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Subsections

Genetic Modification of Food

New findings double the number of bases which build DNA [1]

Four basic units of nucleobases (adenine, guanine, thymine and cytosine) were supposed to build the DNA ruling the genetic code of all living creatures with exception of RNA viruses. Ito et al 2011 report the finding of two other bases, 5-formylcytosine (5mC) and 5 carboxylcytosine (5caC). They are cytosine which was modified by TET proteins. These two molecules are engaged in the demethylation of DNA and stem are active during cell reprogramming. Demethylation caused by these two new bases may reprogram adult cells to become stem cells and cancer.

The researchers also reported that TET proteins can convert 5 methylC (the fifth base) to 5 hydroxymethylC (5hmC)(the sixth base) in the first of a four step reaction leading back to bare-boned cytosine. The reaction with Tet continues on to the seventh and eighth bases, called 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC) from 5mC. Both bases (5fC and 5caC) were found to be active in embryonic stem cells as well as mouse organs and were detected in genomic DNA. The authors concluded that DNA demethylation may occur through TET-catalyzed oxidation followed by decarboxylation. The new findings may be important for cancer research by reactivating tumor suppressor genes that had been silenced by DNA methylation, and may reprogram adult cells.

TET (ten-eleven-translocation) genes [2]

According to Mohr et al 2011 stem cells have the ability to perpetuate themselves through self-renewal and to generate functional mature cells by differentiation. The balance between self-renewal and differentiation must be maintained, otherwise malignant disorders may result. This balance is controlled in part by DNA cytosine-5 methylation catalyzed by DNA methyltransferases.

TET were found to be involved in demethylation by catalysing the conversion of cytosine-5 methylation to 5-hydroxymethyl-cytosine. Members of the TET family are also implicated in embrionic stem cell maintenance and inner cell mass cell specification. Low genomic 5-hydroxymethyl-cytosine and TET2 mutation status was shown in patients with myeloid malignancies and hematological malignancies.


GMO, Genetic Modified Organism, definition

Genetic Modified Organisms according to the Gentechnikgesetz (GenTG)from 20.06.90 (Genetic Technique Law) in Germany are organisms whose genetic material were modified in a way which is not found in nature under natural conditions of crossbreed or natural recombination. The genetic Modified Organism must be a biological unit which is able to multiply itself or to transmit genetic material.
Examples of modifications covered by this law are DNS recombination techniques in which vector systems are used; techniques by which genetic material prepared outside of the cell is introduced directly in the organism. These techniques include microinjection, macroinjection and micro encapsulation, cell fusion as well as hybridization procedures by which living cells are formed with a new combination of genetic material using methods which are not found under natural conditions.

Ployd [3]

Ployd is the number of chromosomes in a cell. The haploid number is the number of chromosomes in a gamete of an individual. This is distinct from the monoploid number which is the number of unique chromosomes in a single complete set.Most of human cells are diploid, that is, they contain one set of chromosomes from each parent. Sperm and eggs contain only one set of chromosomes, they are haploid.

Plants and many amphibian, reptiles and insects contain four sets of chromosomes. This is called tetraploidy. The Monoploid number gives the number of chromosomes present in the cell and euplody is a multiple of the monopliodal number. The human cell has 46 chromosomes which is a multiple of the monoploid number 23. In humans, the monoploid number (x) equals the haploid number (the number in a gamete, n), that is, x = n = 23. In some species (especially plants), these numbers differ.

Commercial common wheat is an allopolyploid with six sets of chromosomes, two sets coming originally from each of three different species, with six copies of chromosomes in each cell. The gametes of common wheat are considered as haploid since they contain half the genetic information of somatic cells, but are not monoploid as they still contain three complete sets of chromosomes from the original three different species (n = 3x).

A monoploid cell is likely to be identical to the cell it was copied from however in haploid cells one of two differing copies of the same chromosome is in the haploid set. Aneuploidy is when the total number is not a multiple of the monoploid number, such as the Down syndrome where there is a single extra chromosome, or there is a missing chromosome in case of the Turner syndrome.

Zygosity [3]

The terms homozygous, heterozygous and hemizygous are used to simplify the description of the genotype of a diploid organism at a single genetic locus. At a given gene or position along a chromosome (a locus), the DNA sequence can vary among individuals in the population. The variable DNA segments are referred to as alleles, and diploid organisms generally have two alleles at each locus, one allele for each of the two homologous chromosomes. Simply stated, homozygous describes two identical alleles or DNA sequences at one locus, heterozygous describes two different alleles at one locus, and hemizygous describes the presence of only a single copy of the gene in an otherwise diploid organism.


F1 hybrid [4]

In agronomy, the term "F1 hybrid" is usually reserved for agricultural cultivars derived from two different parent cultivars, each of which are inbred for a number of generations to the extent that they are almost homozygous. The divergence between the parent lines promotes improved growth and yield characteristics through the phenomenon of heterosis ("hybrid vigour"), whilst the homozygosity of the parent lines ensures a phenotypically uniform F1 generation. Each year, for example, specific tomato "hybrids" are specifically recreated by crossing the two parent heirloom cultivars over again.

Two populations of breeding stock with desired characteristics are subject to inbreeding until the homozygosity of the population exceeds a certain level, usually 90% or more. Typically this requires more than ten generations. After this happens, both populations must be crossed while avoiding self-fertilization. Normally this happens in plants by deactivating or removing male flowers from one population, taking advantage of time differences between male and female flowering or hand-pollinating.

An F2 hybrid is a cross between two F1 hybrids. This second-generation cross does not produce consistent or vigorous plants.


Technology of genetic modification

There are two methods used to introduces a new DNA (gene) in the cell of a plant which is going to be modified:

The "shot-gun" technique

This technique is also called biolistic transformation which was developed by Sanford in 1987.
Cereals are not suitable to be modified by transfection with Agrobacterium tumefaciens and the regeneration of plants whose cells walls were enzymatic digested is very difficult. A device was built to shoot small particles of gold or tungsten against cells. These particles can be coated with DNA material and are so small that they can penetrate cells without lasting damage.
The machine used gun powder and later compressed helium. The particles are accelerated four times sound velocity.
This method is less labor intensive as the Agrobacterium method. The ADN which is being introduced in the host cell is not so complicated as with Agrobacterium and there is also possible to introduce more then 10 different genes at a time. Biolistic transformation can be used to transform all kind of plants, bacteria, moulds, algae and animals.

The transfection with Agrobacterium tumefaciens [5]

This bacterium infects the plant and transfers its DNA to the plant. Agrobacter tumefaciens is a bacterium found in soil. Some strains attack plants transferring a small part or its genetic material to the plant causing tumors.
In the tissue of the tumors Agrobacterium tumefaciens can live and produces there new nutrients (opines).
Opines are products of the condensation of an amino acid and a ketonic acid or an amino acid and a sugars.Examples of opalines are: nopalin(arginine + a-ketoglutaraldehyde ) and octopinOctopin(arginine + sugar piruvate)
Some strains of Agrobacterium tumefaciens have aside of its own ADN genome other plasmides of the size between 200 and 800 kBp (kilobasepairs). These plasmids are responsible for the tumor activity and are therefore called "Tumor inducing Plasmids" (Ti-plasmids).

Ti-plasmides

They carry genes for: The T-DNA is the part of the Ti-plasmids which is transferred to the plant ( Transfer-DNA ) and is limited by the 25Bp repetition as Left Border (LB) 0n the left side and the 25Bp repetition as Right Border (RB) on the right side which are the recognition sequence for the T-DNA.
The transfer of the T-DNA takes place only to wounded plant cells. Certain compounds such as Acetosyringon which are released by wounded cells of the plant act as a recognition for the Agrobacterium tumefaciens in order to link to wounded cells. These compounds are found mainly in dicotyledoneans and only few monocotyledoneans such as asparagus. That is why Agrobacterium can be used only in few cases of genetic modification of monocotyledoneans. With addition off syringon even moulds and important monocotyledoneans can be modified by Agrobacterium tumefaciens.
Special techniques made possible to modify the Ti-plasmids and the T-DNA to avoid the production of phytohormones responsible for the tumor activity, the opine synthesis gene were cut out and gene of the resistance to antibiotic Neomycin and Kanamycin was introduced. The binary vector system uses a big plasmid with the vir-region and a small plasmid with the LB and RB.of an Escherichia coli plasmid.


Protoplasts transformation

Protoplasts are called cells without cell walls.
This method uses pectinase and cellulase enzymes to digest the cell walls of plant tissue.
The vectors used are similar to the method of the shot-gun. The transfer of the DNA to the host is done with the aid of polyethylene glycol or short electrical shocks. This is called electroporation. Selection of the transformed protoplasts an regeneration of the cells is very difficult with this method.
To avoid possible resistance of bacteria to these antibiotics it has been tried to substitute the resistance gene with a gene responsible to the production of isopentenyl transferase which induces a new side shoot of the plant indicating the transformation.

Selection systems

Selection of the transformed cells is being done with antibiotics Kanamycin or gentamycin and neomycin . These antibiotics act toxic on cells without transformation.
The DNA material which is being introduced in a plant must contain four parts:

Marker assisted selection (MAS) [6]

Marker assisted selection (MAS) is indirect selection process where a trait of interest is selected not based on the trait itself but on a marker linked to it.

The gene of interest is directly related with production of protein(s) that produce certain phenotypes whereas markers should not influence the trait of interest but are genetically linked. In many traits genes are discovered and can be directly assayed for their presence with a high level of confidence. However, if a gene is not isolated marker's help is taken to tag a gene of interest.

A marker may be:

- Morphological

First markers loci available that have obvious impact on morphology of plant. Genes that affect form, colouration, male sterility or resistance among others have been analysed in many plant species. Examples of this type of marker may include the presence or absence of awn, leaf sheath colouration, height, grain colour, aroma of rice etc. In well-characterized crops like maize, tomato, pea, barley or wheat, tens or even hundreds of such genes have been assigned to different chromosomes.

- Biochemical

A gene that encodes a protein that can be extracted and observed; for example, isozymes and storage proteins.

- Cytological

The chromosomal banding produced by different stains; for example, G banding.

- Biological

Different pathogen races or insect biotypes based on host pathogen or host parasite interaction can be used as a marker since the genetic constitution of an organism can affect its susceptibility to pathogens or parasites.

- DNA-based and/or molecular

A unique (DNA sequence), occurring in proximity to the gene or locus of interest, can be identified by a range of molecular techniques such as RFLPs, RAPDs, AFLP, DAF, SCARs, microsatellites etc.

A US definition of GMO

"The term "Genetically Modified Organisms" refers to plants and animals containing genes transferred from other species to produce certain characteristics, such as resistance to certain pests and herbicides." In the European community Genetic Modified Organism and its release to the environment are covered by following rules:
1.- Rule 90/220/EWG from 23.04.1990 concerning the release of Genetic Modified Organism in the environment.
2.- Rule 90/219/EWG from 23.04.1990, modified by the Rule 94/51/EWG concerning the use of GMO in closed systems. 3.- Novel Food Decree concerning foods and ingredients which have not yet been used for human nutrition in a worth mentioning extent before. This includes foods covered by the Rule 90/220/EWG; foods initial prepared by means of Genetic Modified Organism but not containing the initial GMO any more such as sugar obtained from genetic modified sugar beet, foods with modified primer molecular structure, foods which had been prepared with or from microorganism, mushrooms or seaweed, foods which had been prepared with unusual techniques which modify significantly the structure of the food.
The content of the German GenTG is similar to the content of the European Rule 90/220 EWG and Rule 90/219/EWG.
The genetic modification of food has the prime score to produce food with:

longer shelf life,
better properties,
using less insecticides in agriculture.

This is true in case of soybeans but Roundup Ready soybean can be efficiently cultivated only with the insecticide of Monsanto. The worldwide insecticides used for soybean will be monopolized therefore by Monsanto

Efforts are being made to breed cereals with better proteins, rape seed with fatty acids better suitable in case of certain diets, other plants missing proteins causing allergies and lactic acid bacteria resistant to virus in the production of milk and meat products thus turning the process of production and the product itself safer.

Please note that the tomatoes on market in Germany are not the same in taste and structure as they had been for years ago, but they have a longer shelf life as before. This however is not due to genetic modification. It is a result of natural crossing of different types of tomatoes.

One should however consider the loss of quality of fresh tomatoes sacrificed on the effort to commercialize the tomatoes and to insure a long transport and a long shelf life.

Unlike to the experiments of Gregor Johann Mendel( 1822/1884 The monk of the Augustiner Abbey who discovered the Mendelschen rules of heredity) modern genetic modification of food introduces alien genes from one species to another completely different one, such as one or more gens of bacteria to the chromosomes of plants. The modern genetic technology interferes deeply with the natural structure of nature.
Crossing plants do not introduce special parts of DNA like terminator gens, marker genes as done by extreme genetic modification techniques.


GMO tomatoes approved by EU Commission

One sort of genetic modified tomato has been approved by the Food Commission of the EU. AstraZenca plc, London has applied for this sort of tomato which has better properties for processing. It is harmless after cooking which denature its proteins and gens. It is therefore not suitable to be sold to consumers which eat the tomato without cooking.

EU guidance on GM animals, plants and derived foodsfood


Guidance on the risk assessment of animal-derived GM food and feed [7]

The EU assessment of GM food and feed, described in Regulation (EC) No 1829/2003 compares GM animals and derived food and feed with the phenotype of traditionally-bred animal. The health status and the physiological parameters of a food/feed producing animal are important indicators of the safety of derived foods/feed

The guidance assumes that traditionally-bred animals have a history of consumption as food and feed, and can therefore, serve as a baseline for the food and feed safety assessment of GM animals, their products and their welfare.

Other topics are addressed, such as molecular characterisation, toxicology, allergenicity of novel proteins, nutritiousness, requirements for the assessment of health and welfare of GM animals bred for food and feed use. Where no comparator can be identified, an assessment of health and welfare of the GM animal itself is considered. The environmental risk assessment of GM animals will be addressed in stand-alone guidance.

EU Guidance on the environmental risk assessment of genetically modified animals [8]

The document, which focuses on GM fish, insects, mammals and birds, outlines the specific data requirements and methodology for the environmental risk assessment of GM animals should applications be submitted for market authorisation in the European Union in the future.

The risk assessment is based on a comparative approach between GM and non-GM animals. The document includes the interactions of the GM animal with target and non-target organisms; environmental impacts of the techniques used to rear or keep the GM animal; and the impact of the GM animal on human and animal health, for example to assess potential risks to farmers, other workers or the general public that may come into contact with GM animals. The draft guidance requires that these and other potential risks are assessed using a systematic, step-by-step approach. Recommendations are also made for the post-market environmental monitoring of GM animals.

According to the Guidance, at present, no applications for market approval of GM animals have been submitted in the EU. However, some non-EU countries regulators are already evaluating GM animals, both in terms of food/feed and environmental safety. The European Commission aims to be prepared for future applications for introduction on the EU market of food and feed derived from GM animals [9]

All stakeholders and interested parties are invited to provide their comments on the guidance through an online public consultation that runs until 31 August 2012.

The proposed Guidance, a loophole for the introduction of animal GM food?

GM animals reared as food are not welcome by the consumer. The European Union should maintain the GM veto instead of preparing for an approval of such food and feed. The recent Guidance is a loophole for the introduction of animal GM food in the EU.


Guidance for food and feed risk assessment of GM plants [10]

The EFSA published on May 2011 an updated guidance for the risk assessment of food and feed derived from genetically modified (GM) plants, assessing developments in areas such as allergenicity and selection of the comparator plant against which the GM plant is compared. The guidance also establishes a new statistical methodology to improve assessment of GM plants.

The guidance outlines methods and approaches for data generation, collection and analysis that must be followed by those submitting applications to EFSA for GM plant risk assessment, focusing on molecular characterisation, field trial design, compositional analysis, toxicological assessment, allergenicity and nutritional assessment. It does not cover the environmental risk assessment of GM plants which is addressed in a separate, stand-alone EFSA guidance document.


Environmental risk assessment of genetically modified plants [11]

This document provides guidance for the environmental risk assessment (ERA) of genetically modified (GM) plants submitted within the framework of Regulation (EC) No 1829/2003 on GM food and feed or under Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms (GMOs).

Labeling of genetic modified food in Europe

The labeling of genetic modified food will be made according to European law EG paper 90/220/EWG. It will be made only in food having the following modifications(EG Paper Nr. 1139/98 and 79/112 EWG)
1.-Food containing alien gens, proteins or DNA .
2.-Food on which the new gens were removed by processing technologies or refining but are changed in some parts so that they cannot be compared with food of nature. An example is GMO rape seed oil with modified fatty acid composition.
Suggested labeling:"Produced with genetic modified corn." In the list, of the ingredients it can be cited "genetic modified" when Soya or rape seed oil is being used.
Food having no alien genes left are not labeled as GMO food ( Genetic Modified Food ). Soybean oil is not labeled as GMO food because refinement removes all modified genes.

Ingredients

Ingredients are also declaration free no matter how manychanges they have undergone (for instance: soybean lecithin is being used as emulsifier in margarine, chocolate and other products )
GMO chymosin, an enzyme used in the production of cheese needs no declaration.
Enzymes are produced by bacteria whose genetic code had been modified,
These enzymes have great utility in the production of monosaccharides such as glucose syrup starting from corn.
No declaration is necessary because no genes or modifications of the resulting product can be demonstrated.One reason to lower the declaration is that meanwhile 90 % of our food would be involved because they had in some way contact with genetic modification.
In October 1999 The Ministers of the EG States agreed with a papers establishing that up to 1% content of Genetic Modified Food are free of declaration.
Refining soybean oil and processing maize and maize starch in Cornflakes for instance high temperatures are needed modifying or destroying the DNA. The GMO-origin cannot be demonstrated with usual methods. These products, even 100% GMO is free of declaration.That is why great retailers gave a ban on Soybean oil because there is no way to be sure to be GMO-free. GMO corn is being introduced in Europe. The resulting corn oil, corn starch, animal feed for cattle in great amount used as silage is free of declaration.
Unilever has announced to label gen modification on food containing GMO soy and corn.
This would include however only food with genetic modified proteins, products with soybean oil are not included.

Compensation in case of GM contamination of organic crops

Denmark is to pay compensation in cases where conventional or organic production has suffered economic losses following genetically modified (GM) material contamination.

The EC ruled that GM contamination could cause economic losses to conventional foods if they have to then be labelled as containing GM material, thus attracting a lower price. Such a scenario could be especially damaging to the fast-growing organic food sector.

EC says that under the current Danish proposals, compensation will be granted only if the presence of GM material exceeds 0.9 per cent and is limited to the price difference between the market price of a crop that has to be labelled as containing GM material and a crop for which no such labelling is required. The Danish authorities will in all cases take action to recover the compensation paid from the farmer from whose fields the GM material has spread.

Switzerland

Switzerland has published a food regulation concerning GMO dated on 1.3.1995 which is mainly identical to the European regulation also here no declaration of Genetic Modified Food is necessary when tho GMO used in the production of the food and the modified genetic material has been discarded.
Meanwhile Unilever UK, Nestlé UK and Nestlé Italy announces that they are going to produce GMO free.
ADM ( Archer Daniels Midland Co), one of the greatest buyer of Soybeans and corn announces to be able to supply Europe with GMO free raw materials. ADM maintains contact with the America n company DuPontwhich makes contracts with farmers concerning GMO crops. It is obvious that the increased efforts to provide GMO free raw materials do have their influence on the price.
The competition between centers of business such as the Far East competing with dumping prices with the European market and even the competition between European countries itself could only be controlled by suppressing the transport facilities increasing worldwide the price of fuel making global marketing so expressive that local ecological isolated markets would be able to survive.
A short way between producer and consumer needs a normal shelf life making unnecessary special efforts to increase it. The great supermarkets however need special packing and other special effects.
As it is impossible to turn back these commercial organizations there is no way out of this dilemma.
Be informed what is going on. This is the only way to protect yourself.Look what is happening in the BSE story, the modification of food, the growing importance of contaminants, such as a possible comeback of DDT the powerful insecticide which deposits in food. Remember however that you are guilty of all this mishaps.
It is the constant desire of consumption that speeds up the possibilities of marketing.
You alone can turn back time by living a life concentrated on true values. Try to find pleasure on the simple things of life.Do not be a slave of savor.

Genetic technique

Are the techniques to promote the transmission of hereditary material between living organism.
Organisms bearing alien genes are called transgene organism (transgene animals,transgene plants or transgene microorganism].There are food and their additives which bear alien genes, other even being genetic modified have lost their alien genes during the manufacturing or refining. The later once are similar to natural products and are therefore free of declaration.
Genetic technique is a very young science:
In 1973 genes where transferred for the first time from one bacterium to another and later on, in 1977 the soil bacterium Agrobacterium tumefaciens was used to transfer alien genes into the cell of plants, or the Bacillus thuringiensis (Bt) has introduced proteins in Novartis Bt 176 maize (bt-Maize) The antisense technique indexAntisense technique was developed in 1990. It suppresses some genes, This was used modifying tomato Flavr and Savr The gene producing ripening enzymes is suppressed and the tomato has a long shelve live
Farming in great scale in USA starts in 1996 with soy, maize, rape seed and cotton.
The European Union allows farming of BT- maize from Ciba Geigy/Novartis/Sygenta.on 1997.

Product producer modification or function Germany
Novamyl Novo Nordisk ( DK) makes flovera better
Europe

Rapeseed

PGS ( B ) sterility/herbicide resistant
Tobacco Selta/Gauloise (F) herbicide resistant
Salad Monsanto (USA) herbicide resistant
Corn Monsanto (USA) herbicide resistant
Great Britain    
Tomato Calgene (USA) retarded ripening
Chymosin Gist Brocades (NL) enzyme for cheese production
Tomato pulp Zeneka (USA) retarded ripening
soybean oil Monsanto (USA) herbicide resistant
Backery yeast Gistbrocades (NL) increased speed of fermentation
Netherlands    
Xylanase Primalcol (SF) better cereal products
Xylanase Quest(Unilever,NL) better cereal products
Novamyl Novo Nordisk (DK) better cereal products
Amylopectin starch..Ayebe (F) additive
Rapeseed oil PGS (B) herbicide resistant
Lipase Unilever (NL) production of certain fatty acids
USA    
Tomato Calgene (USA) retarded ripening
Tomato Monsanto (USA) retarded ripening
Tomato Zeneka (USA) retarded ripening
Tomato NDAPP (USA) retarded ripening
Cotton Monsanto (USA) resistance to insects
Soybean Monsanto (USA) herbicide resistant
Potatoes Monsanto (USA) resistance to insects

Lecithin

Lecithin is a natural emulsifier which is present in Egg yolk and in soy beans.
The world production of lecithin from soy beans is around 180.000 mt.For the production of chocolate 25% were used.The rest was used for margarine, all kind of food and last but not least for drugs and tonics.
USA produces about half of the world supply of lecithin.

Producer Percent of world supply GMO
USA 50 Percent GMO positive
Europe    
from soy beans of USA 17 Percent GMO positive
from soy beans of South America 8 Percent unknown origin
South America    
Argentina 2 Percent GMO positive
Brazil 8 Percent only small parts are GMO free
Asia 5 Percent For home consumption, No export

The slow death of Soy bean

In Germany soy oil is not used any more in the production of human food.Only lecithin is still present in margarine as there is no substitute for it in frying margarine.
Great efforts are being made in research to develop mono- and diglycerides compounds with equal frying anti spatter properties. The efforts to get free of soy results from retailer specifications demanding GMO - free , organic food in order to respond to the desire of the customer which is greatly concerned with the natural origin of food.
The latest great fears of BSE, Dioxins in Food and Genetic modified food created a loss of confidence on the public food control departments.

BSE or dioxins did not modify the genetic code of cattle or hens. The harm caused to the involved species by these problems are turned back as soon as the cause or the epidemie is overcome.
Unfortunately this is not the same in case of Genetic Modified Organisms such as Soy beans.
The change of the genetic code of the plant spreads out through the species.
In few years there will be no natural seed any more present a turn back will be impossible. All benefits of Soy bean such as oil, lecithin,vegetable proteins. soy milk, tofu and a variety of ingredients and foods made from soy bean get lost once for ever in their originality. The aversion to GMO soy bean is strongly present in Germany, UK, France, Australia and many other countries , sometimes hidden by other concerns. Slowly the use of soy bean is dying.
Monsanto is responsible for a loss of confidence in one of the most important agricultural export article of US. It is of main concern of public life to demand that 1.- Soybean planting, transportation, storage, shipping and processing is handled separately from GMO-free Soy bean to restore confidence on the origin of food.
2.- Make a collection of samples of seeds of Soy bean GMO - free in order to guarantee access of future generation to the original seeds. These measures are of urgent importance as the spread of the GMO seed is being forced all over the world by just one profit minded organization damaging the image and the security of an important part of US resources.
These measures should also be extended to corn as seeds from Ciba (Bt-corn, bT= bacillus thuringensis)[12], later overtaken by Novartis are deeply modified in their genetic codes.

Soybean as protein supply for animal feed

Soybean meal which is left from the production of oil is by far the most important ingredient for animal feed. In 1999 the European Union used as animal feed:Million
Soybean meal 26,5 million tons
Sunflower meal 9,1 million tons
Rapeseed meal 5,6 million tons
Pea meal 5,4 million tons
Animal meal 2,4 million tons
Fish meal 0,9 million tons

Animal meal can therefore be substituted by soybean or sunflower. Safety could be increased. The costs of the ban of animal meal as animal feed is insignificant comparing with tho costs to win the battle against BSE.
Soybean meal is not possible to be substituted by other plants. It is therefore important to protect soy plant from genetic modification which might later on turn out to toxic for the plant.

Toxin-free oil rapeseed []

Plants produce defence compounds as a protection against pests. Glucosinolates are such compounds which are toxic not only to pests, but also to animals and humans. According to Halkier et al. 2012, defence compounds are translocated to seeds using two members of the nitrate/peptide transporter family, GTR1 and GTR2, as glucosinolate-specific transporters in Arabidopsis. The authors found that the gtr1 gtr2 double mutant did not accumulate glucosinolates in seeds and had more than tenfold over-accumulation in source tissues such as leaves and silique walls. It is therefore being suggested that GTR1 and GTR2 control the transport of glucosinolates to the seeds of oil plants such as rape. Rapeseed cake, which is a by-product of rape oil, may be used as feed by pig and chicken farmers. Toxin-free oilseed rape could become a feed crop, reducing the dependence of soy cake for animal feed.

The authors try to produce oilseed rape which lack the GTR1 and GTR2 proteins resulting in seeds completely free of glycosinolates.


Identity preserved (IP) non-GM soy bean lecithin and maize

The desire across Europe to avoid genetically modified Foods has lead to an increase of demand for organic and GM-free foods.Austria is the most developed market in Europe. 8.6% of its total farmland is dedicated to organic farming.
Germany and France is expected to develop the greatest market for organic and non-GM foods.
The growing demand of non-GM ingredients for industry creates the need for a certified supply of raw materials. Some bodies which certify organic food chains are Skal Skalin the Netherlands, Naturland in Germany Ecocert in France.
Non-Gm soy bean lecithin is now available with Identity Preserved (IP) non-GM certification coming from the state of Parana,south of Brazil. Non-GM colours such as beta carotene, curcumin, lutein, beetroot and caramelised sugar are available.
Efforts are being made to create xanthan gum derived from GM-free crops of sugar to replace maize derivates because of doubts about the non-Gm status.
Sources of Non-GM, organic, Halal and Kosher status of food ingredients are listed in the Ingrid Database from Fi Data Services, Milton Keynes, UK.

Some of the improvements of food for the future

:

Gluten free cereals

: Gen engineering may produce wheat free of gluten. This aminoacid is not tolerated by sprue patients ( a kind of severe allergy ).

Rice with high level of vitamin A

:In Asia there are regions with undersupply of vitamin A. New seed of GMO rice is rich on provitamin A helping to overcome the deficit.

Transgenetic oil of rape seed

: The GMO oil of rape seed has a modified composition of fatty acids being more valuable then normal oil.

Chymosin

: Is an enzyme obtained from the stomach of calf. It is used in the production of cheese.Because of the prevention of cruelty to animals and because of hygienic aspects many people prefer cheese made with chymosin produced by transgenetic bacteria.

Production of chymosin [13]

The active ingredient of rennet is the enzyme, chymosin (also known as rennin). Rennet from calves are produced from the content newly born calfs abomasum, which is the fourth and final stomach in ruminants which secretes rennet. Calves should be younger than 4 month.

To overcome a shortage of rennet from calves six major alternatives of protease enzymes are on market:

Non-GMO non-calf rennets
  1. Rennet from veal calves
  2. Rennet from adult cows
  3. rennet from pigs
  4. Protease from Mucor miehei
  5. Protease from Endothia parasitica
  6. Protease from Rhizomucor pusillus
  7. Protease from Endothia parasitica
  8. Protease from Mucor pusillus
GMO non-calf chymosins
  1. Chymosins from GMO Escherichia coli
  2. Chymosins from GMO Kluyveromyces lactis
  3. Chymosins from GMO Aspergillus niger)
  4. Saccharomyces cerevisiae.
Other proteases had been proposed as chymosin alternative but not in use,are won from plants:
Cynara candulus, Galium verum, Wrigthia, calycina, Irpex lacteus, Fusarium moniliforme, Sunflower kernel, Calotropis procera.

Advantage of GMO chymosin

Chymosin is now being produced mainly by genetic engineered Saccharomyces cerevisiae. This technology provides chymosine which is identical to the natural chymosin from calf rennet and is being used with few exceptions in the production of all hard cheeses. It is of superior quality, because it has less impurities found in natural non-calf chymosins.

Vegetarians and some religious groups agree to this solution because it avoids killing of calves. [14]

The cheese produced with chymosin from GMO Saccharomyces cerevisiae is considered as GMO-free because the genetic engineered cells of Saccharomyces cerevisiae are not present after the purification of chymosin and the enzyme breakes down during maturation of the cheese. It is therefore not being labelled.

Genetic engineering of yeasts for the production of Chymosin [14]

The DNA encoding the protein chymosin is isolated from calf cells. A copy of this DNA is inserted into plasmids which are then introduced into yeast cells. The genetic modified yeast cells , cultivated in a fermenter vessel, produce chymosin a purer for as found in calw rennet.

Microorganisms which are used in this technology are the yeast Kluyveromyces lactis, the fungus Aspergillus niger var awamori, and a strain of the bacteria Escherichia coli.

Yield in clotting and sensorial changes in ripe cheese using different coagulats [15]

Some reports stressed that there is a higher proteolysis level for microbial coagulants, and with pH decrease, which may lead to a lower yield in clotting and to sensorial changes in ripe cheeses, compared with calf and genetic rennet.

Vasconcelos and colleagues, following these reports, evaluated the influence of pH on yield of milk coagulation with three kinds of coagulants (calf rennet, genetic and microbial coagulant). The analyzed parameters for curd and whey did not differ for the the three coagulants, regardless of milk pH.

Phytase

Is an enzyme used in vegetable food for poultry and pig diets. It liberates the phosphor which is bounded as indigestible phytate. Using phytase the amount of mineral phosphor being added to the food may be reduced and consequently the amount of phosphor which is deposited on the fields.
This turns out to be positive for environment reducing overfertilizing with phosphor. Phytase is produced by GMO bacteria and represents the good side o genetic modification of food.
Other compounds which are produced using genetic modified technologies are: Erythropoietin used in the drug Recormon to treat anemias.
rPA ( recombining plasminogenic activator) used in the drug Rapilysin to treatsever cardiac-infarct.
Monocloned antibodies are also transgenetic biochemical compounds with great chances in future
Other positive genetical products may come soon:
Genetic modified organism like Bacillus subtilis producing enzymes which hydrolyses starch in the production of glucose which is the basis of the production of citric acid and other products.
GMO microorganism will soon produce vitamins such as B1 and B2 as well as aromas and aminoacids such as flavor enhancer and aspartame.
Allergies: It is to believe that the number of allergies is not increasing with the genetic technologies.
The real cause of an increasing risk of allergies is due to a contact to local unknown proteins resulting from globalization of the food marketing by exotic food being imported from everywhere. A recent example is the kiwi allergy and other causes such as environment.

GMO soybean: On the fields of America and Brazil the main producer of soy oil is the seed of " Roundup Ready soybean"being developed by Monsanto.
This seed is tolerant to the herbicide " Roundup ", also manufactured by Monsanto.

The GMO soybean is authorized to be sold in Europe to be used as food for animals and the oil for human food.

In Germany, due to the activity of Green Peace soybean oil is not being used for food.

Low-linolenic soyseeds

Dr Kristin Bilyeu of the USDA's Agricultural Research Service (ARS) has identified the genes in soybeans responsible for the low-linolenic acid trait. Linolenic acid reduces the shelf life and stability of products made from soyoil making it necessary to partially hydrogenate it which leads to the formation of trans-fatty acids. Reducing the amount of linolenic acid in soybeans is a desired breeding objective so that oxidation stable soybean oil can be produced without the production of trans fatty acids. The objective of this work was to determine the molecular genetic basis for soybeans containing 1% linolenic acid in the seed oil fraction and to develop molecular markers specific for identified alleles. Novel combinations of mutant alleles at the three GmFAD3 loci allowed the development of new germplasm containing 1% linolenic acid in the seed oil along with SNP-based molecular markers that can be used in a backcross breeding strategy. Kristin Bilyeu characterized the genes responsible for the linolenic acid formation in the plant, submitting her markers- or identification method- to GenBank, a database of the National Center for Biotechnology Information.

Other low-linolenic soyseeds, the Vistive beans, have been developed by Monsanto.

GMO corn: The GMO corn (called B.t.corn) developed by Ciba-Geigy was authorized to be sold in Europe on the 4.2.1997. The modified corn bears the following gens :
1.- A gene for the production of a B.t.-toxin which protects the plant against a specific insect.
2.- A gene called pat'-gene for the tolerance to the herbicide "Basta " containing Phophosphinotricin. This gene was introduced only to select th plant with the B.t-toxin gene.
3.- A gen called amp'-gene which was introduced in the plant together with the B.t.-toxin gene. It is the ampicillin resistance gene.
The amp' gene produces TEM-1 beta-lactamase which is the most common beta-lactamase found and is responsible for the resistance to ampicillin from 50% of all Escherichia isolated today, from, which 90% are being caused by the RTEM1 type.
The gene is plasmid coded and is denominated as amp' or bla(Tem-1) and exist on a series of cloning vectors such as pBR 322-derivate and pUC- series.
TEM-1 has a low activity against new cephalosporin and may be inhibited by beta-lactamase blocker such as clavulane acid or tazobactame. However under certain conditions there may be created a resistance to amoxicillin / tazobactame as well other combinations of beta-lactame/beta lactamase - inhibitors.
Recently an increase of mutation of TEM-1 and SHV-1 beta-lactamase is noted producing in some cases resistance to new cephalosporin and monobactame.
These derivates are labeled as "extended-spectrum-beta-lactamases" (ESBL), These enzymes were found in Klebsiella pneumoniae, Escherichia coli, Serratia marcescens and other Enterobacteriaceae.
Mutation of TEM1 beta-lactamase such as TEM-30 to TEM-41 may be the reason why the inhibition through clavulane acid is very low.
Bush 1995 has introduced therefore a subclass labeled as "2 br" for these variants.
The "inhibitor resistant TEM-beta-lactamases" ( IRT) was found only in Escherichia coli and in certain Proteus mirabilis and Klebsiella pneumoniae.It might happen that IRT also spreads to Haemophilus influenza and Neisseria gonorrhoeae having frequently TEM-1-beta lactamase.
The use of ampicillin to treat Enterococcus and Haemophilus influenzae infections should be made nowaday preceded by a sensibility test and using suitable beta-lactamase blocker by necessity.
The possibility of a transfer of gens from B.t. corn from Ciba to bacteria cannot be completely excluded.
To transfer the amp'gene to a bacterium it is necessary that the plant releases the amp' gene together with the "origin of replication" (ori) from pUC without damage. The plant cell usually degrades DNA during release with his own nucleases.
DNA enzymes in the paunch and the digestive tract from mammals and in the soil where bacteria may grow also destroy the gens.
The ability of bacteria to take over alien gens is seldom. Only very few bacteria have this ability, together nucleases found everywhere the probability of a transfer of the amp'gene from corn to a bacterium is very low.
The formation of a replicon in the cell, as for example the binding of the extremities of a DNA - fragment could only happen through an illegitimate cross over.
The host place of the origin of a replication exists only in a very limited number of Enterobacteriaceae.
All the above mentioned restrictions make the possibility of a transfer of amp' gene from plant to a bacterium seem to be unlikely.
It is supposed that all human beings have an amp' gene bearing Escherichia coli in their intestinal tract without having been exposed to beta lactame antibiotics. Approximately 50% of all clinically isolated Escherichia coli are already resistant to ampicillin of which 90% have a TEM-1 beta-lactamase.
The amp' gene already being so frequent there is no further danger to increase significantly the number of resistance by a transfer of the gene from Ciba corn to a bacterium.
However the alien gene has no function in the new corn. In future developments it is important to avoid marker gens with resistance to antibiotic or herbicides.

Cellular mechanisms generating chromosomal breaks in bacteria such as Escherichia coli [16]

Nudler et al. 2011 explain how bacteria generate mutations adapting to stressors like antibiotics. The study, using Escherichia coli as living cells, assesses the interaction of replication, transcription, translation and DNA repair with the big actors: replisome, a protein complex responsible for DNA synthesis, RNA polymerase, an enzyme responsible for RNA synthesis, and ribosome, a molecular structure responsible for protein synthesis. Collisions between replisome and RNA polymerase occur frequently in cells because both share the same DNA track, but the speed of the replisome is much faster than that of RNA polymerase.

In this study the co-directional collisions were found to lead to DNA double strand breaks (DSBs) or mutations. Importantly, however, such DSBs appear only if the replisome collides with backtracked RNA polymerase, head-on collisions do not produce such breaks.

Backtracking of RNA polymerase along RNA and DNA, is an intrinsic property of all cellular RNA polymerases from bacteria to humans. Multiple anti-backtracking mechanisms that employ various transcription factors exist in bacteria and nucleus-containing cells, including human cells. The authors stress that the interaction between translating ribosomes and RNA polymerase is important to keep genomic stability because it prevents backtracking. A mechanistic model is proposed to explain such backtracking.

Ribosome functions as sensor of cellular metabolism and stress, such as starvation or antibiotics resulting in mutations. Such mutations depend on error-prone DSB repair resulting in adaption to the environment or other stressors. The backtracking-based mechanism of DSB drives, therefore, the stress-driven evolution in bacteria. The organization of replisomes and RNA polymerase is preserved in evolution. Backtracking-driven genome instability for E.coli may, therefore, occur also in other organism and may explain certain human diseases,


Detection of GMO in food

There are many genetic modified foods on market. To supervise the declaration new methods of analysis were needed which are based on molecular biological principles.
Detection can be made looking for the new specific protein or detecting the new genetic material.
The detection of GMO is very difficult because there are so many other compounds which may interfere in the detection, such as polysaccharides which can inhibit the polymerase chain reaction leading to false negative results.
The food processing causes a denaturation of the proteins on the DNA which is being on test being responsible for failing to be recognized by primers and antibodies. The tests should therefore be suited for the specific processing methodes used. The genetic modified material is often present in very small amounts. Sometimes the transgenetic protein is located in other parts of the plant and the part which is being used as food has no or very little transgenetic material such as the Bt toxin which is present in leafs but not in maize kernels of Novartis BT 176 maize. Usual methodes of GC-MS, HPLC and capillary electrophoresis are unable to detect them. Immunological detection of the transgenetic proteins such as Western Blot or ELISA are now used.
The most effective method to determine transgenetic material is to amplify the alien sequence of the promoter and the gene.
The analytical methods contain the following steps[17]:
1.- Extraction of DNA:It is necessary to extract the genetic material free from other impurities which might interfere in further steps of the analysis.
2.-PCR reaction ( Polymerase Chain Reaction)
The PCR reactions are suited to multiply and amplify specific fragments of DNA that are alien genes to the food being analysed. The primer starter molecules used in the beginning of the reaction decides which sequence of DNA will be multiplied. To avoid false negative results due to inhibit action of impurities during extraction of the DNA it is important to include a positive reaction.
3.-Making the PCR product visible
Through gelelectrophoresis (agarosegelelectrophoresis). The products of the PCR reaction can be made visible together with the determination of the length of the base pair, the alien gen.
4.-Confirmation of the results
The confirmation of the results are being made by controlling the sequence of the base in the PCR product using specific sequence restriction, hybridization with specific sonde
Nested PCR and
Sequencing
The basic PCR gives only qualitative indications. To obtain quantitative results the Competitive PCR or the RT-PCR should be used.


Competitive PCR

Is not so expensive as RT-PCR but there are dilutions to be made which take much time. If two sequences are present with the same complementary DNA sequence for the primers to annel they compete for binding of the primers. A DNA sequence ( internal standard) which is much shorter as the target DNA The amplification products can be separated on a agarose gel. The fluorescence of these products is proportional to the amount of amplified DNA. There are two bands on the agarose gel. Dilutions must be done until the brightness of both bands are equal.


RT-PCR Real-time PCR

The amount of molecules produced during each stage is measured rather then at the end as happens with competitive PCR.
To demonstrate the Presence of modified DNA the PCR-Method is today favored. Other Methods are the gelelectrophoresis sequencer and ELISA .
These methods are used as well in food chemistry as well in clinical researches because they are based on the same principles.

Reliability of real-time reverse-transcription PCR in clinical diagnostics [18]

Real-time PCR has played a decisive role in the sequencing of the human genome, comprehensive genomic, mRNA and miRNA expression profiling of many diseases, detection of human pathogens, diagnosis and prognosis, treatment monitoring and transplant biology. Murphy and Bustin 2009 cite, however, some technical deficiencies such as the demonstrated through its association with the measles, mumps and rubella vaccine/autism controversy. The authors call for careful experimental design, validation and analysis.

The need of gidelines for qPCR data publications [19]

According to Bustin 2009 real-time quantitative PCR (qPCR) due to ill-assorted pre-assay conditions, poor assay design and inappropriate data analysis methodologies resulted in inconsistent or misleading publication. Materials and methods informations are often insufficient for an evaluation of presented data. A set of guidelines of minimum standard for the provision of information for qPCR experiments ("MIQE"). These guidelines may improve the reliability of qPCR nucleic acid quantification technology.

Data normalization and reference genes for RT-qPCR [20]

Galiveti and colleagues 2010 report that numerous non-protein-coding RNA (npcRNA) molecules form a class of untranslated RNAs with significant biochemical activities. Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) lack appropriate internal controls in the analysis of npcRNA. The expression of protein-coding reference genes, also termed "housekeeping" genes (HKGs) vary among tissues and different experimental conditions and are, therefore, questionable as reference in npcRNA expression analyses.

The authors determined the most suitable internal control with least expression variance. They found that five npcRNAs presented better expression levels in different tissues than common HKGs. The authors termed these genes as housekeeping RNAs (HKRs) These genes may be used for RT-qPCR data normalization in human transcriptome analysis, and might also be used as reference genes.

The Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines [21]

Bustin and colleagues 2009 stress the lack of consensus on how to perform and interpret quantitative real-time PCR (qPCR) experiments. To improve the reliablility and the interpretation of qPCR results the authors developed the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines. These guidelines describe the minimum information together with a checklist which should be included in the manuscript submitted to publishers. Data such as disclosure of all reagents, sequences, and analysis methods are essential for the work of reviewers which assess the validity of the protocols. These guidelines aim to improve the reliablility and the interpretation of qPCR results.

Quantification of mRNA using real-time RT-PCR [22]

Nolan, Hands and Bustin 2005 describe a series of RT-qPCR protocols that illustrate the essential technical steps required to generate quantitative data that are reliable and reproducible.

New sensor turns forensic DNA analysis faster and less expensive than PCR [23]

Roy and colleagues 2009 describe an electrical nanogap sensor for the detection and quantification of DNA. The device is based on the electronic transduction mechanism. It uses standard silicon microfabrication technologies.

A pair of micro-sized metal electrodes separated by a nanogap, in combination with special chemical probes capture segments of DNA immobilising them, followed by hybridization with target strands and metallization. The resulting change in conductivity detects and quantifies the targeted DNA.

The new technique is faster, less expensive, and more practical than DNA detecting methods which use the polymerase chain reaction (PCR).


Gene delivery systems using nanotechnology

Gene delivery is the process of introducing foreign DNA into host cells. Gene delivery is, for example, one of the steps necessary for gene therapy and the genetic modification of crops. There are many different methods of gene delivery developed for a various types of cells and tissues, from bacterial to mammalian. Generally, the methods can be divided into two categories, viral and non-viral.

Viral methods: Virus mediated gene delivery utilizes ability of a virus to inject its DNA inside a host cell. A gene that is intended for delivery is packaged into a viral particle.

Non-viral methods: Non-viral methods include physical methods such as microinjection, gene gun, impalefection, hydrostatic pressure, electroporation, continuous infusion, and sonication and chemical, such as lipofection. It can also include the use of polymeric gene carriers (polyplexes)

Non-viral gene delivery system for soft tissue engineering [24]

Saul et al, 2007 presented a non-viral gene delivery system to integrate polymeric non-viral gene carriers (polyplexes) within a novel three-dimensional, sphere-templated fibrin scaffold suitable for soft tissue engineering applications.

The study was performed by seeding scaffolds with NIH-3T3 fibroblasts. Different transgene expression profiles were achieved based on the spatial distribution of polyplexes within the scaffold. Surface-coated polyplexes achieved one order of magnitude greater expression than polyplexes embedded within the scaffold, report the authors.

Nanoparticles as gene vector [25]

A new gene vector, cationized Lycium barbarum polysaccharides (cLBP) nanoparticles, prepared with different amine compounds for non-viral delivery of plasmid DNA encoding for transforming growth factor beta 1 (TGFbeta-1) was developed by Wang et al 2011. Transfection is the process of deliberately introducing nucleic acids into cells. The authors stress that cLBP presented low cytotoxicity and the transfected cells had high cell viability.

Gene delivery of DNA polyplexed with nanoparticles for stem cells [26]

A non-viral gene delivery system using nanoparticles was developed by Park et al 2010. for gene delivery to human mesenchymal stem cells (hMSCs). These stem cells are used in cell differentiation applications and clinical therapies. Using polyplexing with polyethylenimine, the authors could improve the cell-uptake ability of the nanoparticles.

Biodegradable poly (DL-lactic-co-glycolic acid) (PLGA) nanoparticles polyplexed with polyethylenimine (PEI) enhanced the cellular uptake of SOX9 DNA to promote gene expression in hMSCs stem cells, report Kim, Park and others in 2011. [27]

Contamination with mouse DNA leads to false results in XMRV retrovirus, is forensic genetics affected? [28]

Urisman and colleagues 2006 describe the presence of XMRV retrovirus in prostate cancers. [29]
Mikovits and colleagues 2009-2010 reported the findings of the XMRV virus although in chronic fatigue syndrome patients and advocate the use of more than one type of assay in order to determine the frequency of XMRV infection in patient cohorts in future studies of the relevance of XMRV to human disease. [30] [31] [32]

Recent reports on gammaretrovirus, XMRV (xenotropic murine leukemia virus-related virus), found in patients with chronic fatigue syndrome (CFS) may be false because of contamination of the human DNA sample with mouth DNA. Oakes and colleagues 2010 write that at least four groups could not detect the virus in CFS patient, using another methodology. The authors point to the high degree of similarity between XMRV and endogenous MLV proviruses, and mouse cells contain more than 100 copies each of endogenous MLV DNA. Oakes and his team urge to distinguish contaminating mouse sequences from true infections. [28]

Increased efforts must be done to avoid false results in genetic researches using highly sensitive PCR technology wch are also used in forensic genetics were false results may convict innocents.

Hoh and colleagues 2010 report that several subsequent studies failed to detect the virus in patients suffering from CFS. All sequences by nested PCR and cultures for XMRV virus remained negative in German CFS patients. [33]

Van Kuppeveld and colleagues 2010 report that the XMRV could not be found in chronic fatigue syndrome CFS patients in the Netherlands. The authors doubt on the claim that XMRV with CFS. }citeKuppeveldxenotropicmurine

Switzer and colleagues 2010 did not find any evidence of infection with XMRV in U.S. population of CFS patients or healthy controls by using multiple molecular and serologic assays, and could not support the findings of Mikovots and colleagues. [34]

Groom and colleagues 2010 did also confirm absence of the of xenotropic murine leukaemia virus-related virus in UK patients with chronic fatigue syndrome. [35]


Western Blot

The method of Western Blot the extraction of the transgenetic protein from the food is done by means of a nitro-cellulose membrane which binds the proteins. The membrane is immersed in a solution of a specific antibody together with an enzyme resulting in a colour reaction. This method is very labour intensive and therefore not being used in routine.


ELISA (Enzyme Linked Immuno-Sorbent Assay

It is based on the same principles used for Western Blot. The membrane is substituted by a plastic plate with 100 and more wells being therefore suitable for many tests at the same time.

Round Up Ready Soybean,Monsanto Co

The Round Up Ready Soybean is a glyphosateresistant soybean (Glycine max). Glyphosate inhibits the enzyme of the metabolism of aromatic aminoacids in plants, the so called EPSPS 5-EnolPyruvylShikrimi-3-Phosphate-Synthesis.
The gene of glyphosate tolerance comes from Agrobacterium tumefaciens, strain CP4.
The transferpeptid as carrier of the EPSPS in the chloroplast comes from Petunia hybrid (transit-signal-sequence)
The detection of GMO Soybean Round Up Ready is made using the specific primer pair B1/B2 which couples with the promoter sequence CaMV 35S and with the transit - signal- sequence,
This primer pair gives a PCR-product of 172 Bp which hybridize with the DNA- sonde H-35s-ar1 after the transfer to the membrane [36].The range of options of PCR analysis of genetically modified organisms (GMOs) is expanding from day to day. As there is a great variety of commercialized GM plants grown in USA and in Europe, being exported all over the world , laboratory work is getting always harder to detect all possible GMOs as reference materials are not always attainable.

Risks of genetic engineering

: According to a statement of Prof Wolfgang van Daelen, WZB, Berlin 1997 today there are no empirical or plausible theoretical arguments that genetic modified food represent a greater risk to the consumer as it is with normal food.
Risks can never be completely eliminated, however there are no real risks known until present date. Future research to avoid risks of genetic engineering will be handled in two ways:

  Proactive risk research: This way asks what can happen ?
  Monitoring: The supervision by experts of the fields and the processing of food.

The greatest security in genetic engineering is hoped to be attained with use of both ways.

Allergy and soybean [37]

Why such a trouble about genetic modified soybean ?
Soybean was the first genetically modified plant to be introduced in widespread agriculture.The approval of the new plant by the Food and Drug Administration in USA (FDA) is made by determining the allergenic potentials of the plant.
For this purpose the FDA has established in 1992 a guideline to determine the allergenic potential of a plant.
In 1995 followed the guideline of OECD and WHO to determine allergic components in plants.This guideline contains the principal ideas of the guideline of 1992.These guidelines have three main points: The alien gene from GMO Soybean produces the enzyme CP4 EPSP-Synthase.This enzyme was compared with existing allergens.

Size of the molecule

Only the size of the molecule was identical to known allergens.This is however not relevant because the molecule is inactivated by heat.Processed Soybean meal is therefore not allergenic because of this heat barrier.

Digestion

The CP4EPSP-Synthase molecule is not resistant to digestion, being destroyed in the stomach within 15 seconds.

Bounding to long chained sugars

Typical allergens are bound to long chained sugars , the new enzyme does not have sugar in his molecule.

Degree of strangeness

The degree of strangeness is an important element of comparison with other allergens.The new enzyme CP4EPSP-Synthase is similar to EPSP - Proteins from baker yeast and indexBacillus subtilis Bacillus subtilis.Both of them are considered as GRAS (Generally Regarded as Safe)

Amount of Allergen in the final food

The amount of CP4 EPSP-Synthase in the final food is so small that only 0,001% in total and 0,08% in the proteins of the food are found. Allergenic substances are usually the major part of the food.

GMO soybean oil and allergenity

There are no allergic reactions known from refined oil of GMO soybean.The modified allergenic residues which are left after refining are below 1 ppm.
CP4EPSP protein and Round Up Ready Soybean is therefore considered as not allergenic.

French position to GMO [38]

Some environmental organization like Agir pour l'environnement, Greenpeace, France nature environment combatte the GMO.
Some organizations like the FNPL (Federation Nationalle des Producteurs de Legumes) and the CP (Confederation Paysanne) are against transgene food.
These Organizations are united to fight health- and environment- risks. According to them everything being resistant to antibiotic should be forbidden. Other Organizations like the FNSEA and the CNJA support GMO when:
  1. Possible health- and environment risks are eliminated,
  2. An acceptable solution concerning labeling of GMO food is found,
  3. The national agriculture and the distribution system of food is protected against distortion of trade.

Recall of GMO maize seeds in France

In the end of April 1999 there was a recall of maize seeds in France (Elsace) after being proved that the seeds were genetically modified.The recall was confirmed by Pioneer Seeds company.In Germany there were also US GMO maize seed found. DER BUND (German environment organization) says that it should be avoided that frontiers between genetic modified food and natural grown food are erased. The consumer should keep the right to choose between both foods.[39]
Only Germany and Spain have authorized the use of genetic modified maize. All other member s of the EU do not accept it.

PTI: More unmodified Soya proteins

Protein Technologies International (PTI) most important producer of soya proteins, guarantee that it can supply the demands of genetic unmodified proteins from soybeans in Germany. According to PTI it is a fairytale that US soybeans are not separated in GMO and unmodified. To make sure no modified GMO is being used PTI has developed an Identity Preservation Program which is similar to the "Cert ID" scheme for unmodified foods recently introduced by the trade companies (April 1999).

US exporters want to supply demands of GMO-free Soybeans and cereals

The exporter and cereal specialist Northland Seed and Grains in Minnesota guarantee its products to be 100% GMO-free (April 1999)[40].
Two great cereal buyers accept only grains which can also be sold in Europe.

Unilever UK :Renunciation to genetic modified ingredients

Van den Berg Foods UK and Birds Eye Wall's (part of Unilever UK) will stop the use of GMO ingredients changing the with alternative unmodified ingredients, what also includes GMO-free Soy products (April1999). The efforts to produce GMO-free products show that there is a growing market for these products.
Meanwhile Monsanto introduces for autumn 1999 the GMO Roundup Ready Soybean in Brazil
The industry likes the"Brave New World" from Aldous Huxley.Rhone-Poulenc,a chemical giant and Limagrains producer of seeds united to create the Rhobio Company being responsible for biotechnology activities.
It has even been tried to change "Geneticaly modified " to " Geneticaly improved ".
The Nestlé Group refers to rising prices on GMO-free products.
The food dealers like Carrefour, Casino, and Systeme U make sure to get guarantee as GMO-free from the producer.It has shown that it is almost impossible to guarantee GMO-free products because basic components of food are already modified.Carrefour sells products with GMO-free guarantee. The Danone company , Paris announces not to use GMO - raw materials in their production in Europe.
The dealers are afraid that customers guided by dubious publications can make their own buying decisions to get out of control.
The dealers support therefore the labeling of GMO. Other supermarket chains in Great Britain which took out of their shelves products with genetic modified ingredients including meat from animals which had been feed with GMO animal feed around July 1999: Sainsbury, Safeway, Marks and Spencer, Northern Foods, Unilever and Nestlé GB as well as Nestlé Italy. Sainsbury searches for GMO-free cereals for animal feed to produce GMO-free meat and poultry [41].


Terminator-Gen

In the EU there are rules demanding farmers to pay licenses to seed breeders when seeds from the last crop are held back to be reused once again. The seed breeders say that they are loosing much licenses in Austria where these rules are still not introduced.
USA breeders are therefore working to introduce a new gene in their seeds, the so called "Terminator Gene" which lets seeds germinate only one time.There is no use to keep seeds for the next time. Farmers have to by their seeds from the breeder. If the terminator gene spreads out in free nature all plants will stop to germinate. The use of genetic techniques should not be allowed to collect fees or other financial purposes.[42]
Monsanto has made its golden rice available free of charge to developing countries. This rice had been modified with genetic engineering being enriched with vitamin A. The rice could prevent millions of cases of blindness caused by vitamin A deficiency. Monsanto hopes to sell the specific agro chemicals for this plant.


Terminator biotech technology

The Terminator technology or Genetic Use Restriction Technologies (GURTS), sterilises seeds in order to force farmers and gardeners to buy new seed each season. [43]

The Food Commission of UK argues that Terminator seed will prevent farmers around the world saving their own seed, endangering seed security, the environment and consumer choice. The United Nations Convention on Biological Diversity (CBD) launched a moratorium on the field testing and commercialisation of terminator technology, agreed in 2000.

Australian, New Zealand, US and Canadian governments tried to undermine the existing moratorium at the CBD meeting in Granada, Spain, arguing that the technology would increase productivity. It was also recommended that CBDs precautionary approach should be replaced by case-by-case risk assessment.

Greenpeace today called upon the 188 states at the 8th meeting of the UN Convention on Biological Diversity (CBD) in 22. March 2006 in Curitiba, Brazil, to maintain the moratorium on the field trials and commercial releases of Terminator seed technology which was agreed six years ago. [44]

Governments at the United Nations Convention on Biological Diversity (CBD) have unanimously upheld the international de facto moratorium on Terminator technology at the 8th meeting of the CBD in Curitiba, Brazil. According to Maria Jose Guazzelli of Centro Ecológico, a Brazil-based agro-ecological organization the Convention rejected the efforts of Canada, Australia and New Zealand - supported by the US government and the biotechnology industry - to undermine the moratorium on suicide seeds. [45]
Luxembourg, Greece and Austria consistently vote against standard GMO approvals. New patents for Terminator technology have already been granted in Europe and Canada, and applications have been submitted in China, Japan and Brazil.

Hybrid rice

The FAO maintains the International Rice Commission IRC which coordinates international activities to increase rice yields:

Hybrid rice for Egypt [46]

The International Rice Commission of FAO indexInternational Rice Commission selected rice from more than 200 hybrid varieties to help Egypt to produce more rice with less water and less land, achieving the worlds highest national average rice yield in 2005. C4 rice [46]
Converting rice from a C3 plant to a C4 plant, where the "C" refers to the carbon captured by photosynthesis for growth as land and water resources available for rice production keep diminishing as a result of urbanization and industrialization.

However, according to the International Rice Commission Secretary Nguu Nguyenit, in 25 August 2006, Rome, it will take several more years before the C4 rice varieties may become available. And, then it has to be made sure that they are safe for human and animal consumption as well as for the environment.

Concerns related to biosafety, conservation of rice genetic diversity, intellectual property rights and access are risen. The International Rice Commission believes that international capacity building is urgently required to ensure that new innovations benefit local people and do not incur long-term costs to the environment.

In the medium term, increasing rice production in some countries could require a different approach, one based on introduction of better crop management practices. The results from pilot tests in developing countries since 2000 have demonstrated that very high yield with existing varieties can be obtained with improved crop management (ICM).

Improved Crop Management ICM and increasing rice yelds [46]

In the Philippines FAO's Improved Crop Management ICM has given excellent results increasing rice yields in test regions setting planting dates to expose crops to higher solar radiation, optimising seeding density, balanced plant nutrition, careful water management and the introduction of newly-developed hybrid varieties such as SK 2034 and SK 2046.

GM rice in Japan [47]

C4 rice

C3 plants which are less photosynthesis efficient include sugar beet, rice and potatoes. C4 plants highly efficient in the use of light for their biosynthesis are maize and sugar cane.

Genetically modified rice in Japan

[47] Professor Joe Cummins in his review, points out some overlooked dangers of genetic engineering of rice in Japan.

The C4 rice

Bioengineering tried to modify rice introducing a transgene for that enzyme from maize which expresses the enzyme phosphoenolpyruvate carboxylase (PEPC) fixing carbon dioxide in C4 plants. C3 plants lack this gene and fix carbon dioxide exclusively through an enzyme called Rubisco in the chloroplasts. A hygromycin resistance marker was added to the PEPC gene in the C4 rice, but photosynthesis did not improve.

New trials are ongoing with the gene for the enzyme phosphoenol pyruvate carboxylase (PCK) from Urochloa panicoides (liver weed).

Dwarf rice

Dwarf rice resists to lodging in wind and rain. A gene was introduced in rice to degrade the hormone gibberellin controlling height of the plant. This gene did suppress seeding, reducing yields.

The rice was then transformed with the hormone-degrading gene and put under control by tissue specific promoter for gibberellin synthesis, and terminated with tnos, together with a hygromycin resistance marker. There are no final results published.

Rice for alkaline soil [47]

To avoid iron deficiency in rice, when planted on alkaline soil, a gene which expresses the enzyme nicotianamine aminotransferase (NAAT) in barley was transferred to rice. This enzyme from the root of the plant, in alkaline soil, releases molecules called phytosiderophores turning iron uptake possible. Enhanced iron uptake under these conditions, improved yiels of GM rice.

The NAAT gene transfer was achieved with a CaMV promoter and terminated by tnos, and accompanied by hygromycin-resistance and neomycin-resistance marker genes

bacterial blight control [47]

To control rice bacterial blight the gene for the expression of cecropinB peptide, as strong antimicrobial of the larvae of the silk moth, Bombyx mori, was introduced into rice. A very complicated construct was designed using a version of the CaMV 35S promoter with enhancer 5p, the omega sequence from tobacco mosaic virus followed its promoter and the first intron of a phaseolin gene, a rice chitinase signal peptide, the cecropin sequence, terminated by tnos and a kanamycin-resistance marker.

Insect resistant rice [47]

Stem borer insect resistance

To enhance resistance against stem borer insects a trypsin-inhibitor, interfering with the digestion of the stem borer was introduced into rice, following a very complicated system: A synthetic trypsin-inhibitor gene derived from the winged bean with a reduced GC (guanine-cytosine) content to improve messenger RNA production, added to an enhanced CaMV promoter , together with a tobacco mosaic virus omega sequence and the first intron of a phaseolin gene, and terminated with tnos. In addition, a hygromycin resistance marker was also inserted.

Army worm larvae

The army worm larvae became susceptible to the common soil baculovirus when ingesting the pox virus gene product of GM rice. A synthetic insect pox gene with an altered DNA sequence was introduced in the rice using a CaMV promoter, a non-coding region of the rice stripe virus RNA and terminated by tnos, together with a hygromycin resistance marker. The natural resistance of the army worm larvae against the baculovirus is destroyed when it comes to a contact with the transgene DNA of the rice.

Biosecurity concerns relating GM rice in Japan [47]

Pofessor Joe Cummins concludes that the human cytochrome p450 genes should not be used in rice because they activate carcinogens. Cummins refers to the possibility the human genomes to be invaded by trangene DNA, through illegitimate and homologous recombination, following an extensive use4 of the aggressive CaMV-based superpromoters, and the incorporation of human genes. Dangerous consequences include the creation/activation of new viruses or cancer.

Unauthorised US GMO rice Bayer ll Rice 601not allowed to enter the EU [48]

The European Commission in August 2006 has adopted a decision requiring imports of long grain rice from the USA to be certified as free from the unauthorised GMO LL Rice 601. This unauthorised GMO had been found in samples of commercial rice on the US market. The emergency measures adopted by the Commission today mean that, with immediate effect, only consignments of US long grain rice that have been tested by an accredited laboratory using a validated testing method and accompanied by a certificate assuring the absence of LL Rice 601, can enter the EU.

Member States authorities are responsible for controlling the imports at their borders and for preventing any contaminated consignments from being placed on the market. In addition, they should carry out controls on products already on the market, to ensure that they are free from LL Rice 601.

Business operators importing rice from the USA also have responsibility for ensuring that LL Rice 601 does not enter the EU food chain and that imports are certified as free from this unauthorised GMO, in accordance with the EU food law principle that operators are responsible for the safety of the food or feed that they place on the market.

The Bayer long grain LL Rice 601 review of incidence handling [49] [50]

The FSA will review its handling in the case of the Bayer long grain LL Rice 601 in 29 November 2007. A judicial review in February 2007 found in favour of the regulatory agency, but unveiled a number of mistakes made by the Agency to avoid any further spread of the GM rice.

GM rice in USA is widespread. The crossover to crops scheduled for export to Europe is highly possible. The European control system is therefore to be strengthened, because the US is one major supplier of EU with long corn rice, followed by India, Thailand and Guyana.

Lawsuits against Bayer [51]

The EC imposes no penalties and costs against Bayer.

Bayer is being lawsuited by rice farmers from Arkansas alleging that the corporation failed to prevent Liberty Link (LL) rice 601 GM rice from entering the food chain. As a result of the contamination of rice crops the price of US longcorn rice felt considerably

Commercial rice in the United States was found contaminated with genetically engineered (GE) Liberty Link (LL) rice 601, produced by Bayer. Liberty Link (LL) rice 601 was developed to tolerate the herbicide, glufosinate ammonium. Imports were, as a result, immediately banned in Japan.

Greenpeace International calls on the EC to stop reacting to contamination 'accidents' and start preventing them instead. The EC should identify countries and products that are at high risk of contaminating our food supply with illegal or dangerous GE organisms and implement screening, preventative testing and, where there is no demonstrated capacity to prevent contamination, total bans.

Greenpeace International calls on other major importing regions such as the Americas, Africa and the Middle East to take similar steps immediately until the US can guarantee that their rice supply - and other foods - are no longer contaminated.

There is currently no evidence that LLRICE601 has been imported into Canada.

In advance of the announcement, the CFIA and Health Canada, in consultation with the USDA and Bayer CropScience, conducted preliminary assessments of the risks that this incident posed to Canadian food and feed and the environment. The assessments determined that it is unlikely that low levels of LLRICE601 pose a risk to human health, livestock or the environment.

Canadian assessment determined that there is no risk coming from low levels of LLRICE601 [52]

LLRICE601 is genetically very similar to another type of genetically engineered rice ( LLRICE62) which has been thoroughly assessed and approved for feed and food use in both Canada and the US However, this line of rice is not used commercially in Canada and the US While there is currently no evidence that LLRICE601 has been imported into Canada, the Government of Canada continues to work with the USDA to actively gather information about the distribution of this product.

U.S. Food and Drug Administration's Statement on Report of Bioengineered Rice in the Food Supply [53]

The bioengineered variety of rice, called LLRICE601, expresses the phosphinothricin - N - acetyltransferase (PAT) protein which provides tolerance to glufosinate-ammonium herbicide. This rice variety, not intended for commercialization, was not submitted to FDA for evaluation under the Agency's voluntary biotechnology consultation process. However, crops containing the PAT protein have previously been evaluated for safety by FDA on a number of occasions through the Agency's voluntary biotechnology consultation process.Bayer has informed the Agency that LLRICE601 is present in some samples of commercial rice seed at low levels.

In addition, Bayer has provided information about the safety of the PAT protein, molecular characterization, and nutritional composition of grain from LLRICE601. Based on the available data and information, FDA has concluded that the presence of this bioengineered rice variety in the food and feed supply poses no food or feed safety concerns.

Liberty herbicide and NAG (N-acetyl-L-glufosinate)

Smith glufosinate Jeffrey M. Smith says that Liberty herbicide (also marketed as Basta, Ignite, Rely, Finale and Challenge) can kill a wide variety of plants, bacteria, fungi and insects, and has toxic effects on humans and animals. The herbicide is derived from a natural antibiotic, which is produced by two strains of a soil bacterium. In order that the bacteria are not killed by the antibiotic that they themselves create, the strains also produce specialized enzymes which transform the antibiotic to a non-toxic form called NAG (N-acetyl-L-glufosinate). The two genes which produce these enzymes are inserted into the DNA of GM crops.When the plant is sprayed, Liberty's glufosinate ammonium is absorbed by the plant where the enzymes convert it primarily into NAG protecting itself against the weed killing poison.

NAG, accumulates in these GM crops. NAG may be re-transformed back into the toxic herbicide inside our digestive system by gut bacteria, primarily found in the colon or rectum.
The conversion of NAG back to glufosinate was found to be up to 10% and one-third in goats.

Jeffrey M. Smith says it is believed that although these parts of the gut do not absorb as many nutrients as other sections, rats fed NAG did show toxic effects. A goat study also confirmed that some of the herbicide regenerated from NAG ended up in the kidneys, liver, muscle, fat and milk.

Low chances of consuming significant amount of NAG [54]

Christopher Preston of the University of Adelaide, Australia counters the affirmations of Jeffrey M Smith in AgBioWorld in 30.08.2006:
"Transgenic glufosinate resistant plants do convert glufosinate into NAG; however, very little if any NAG ends up in the grain and none in processed foods." "The chances of consuming any significant amount of NAG are very low."

Preston concludes: "It is true that transgenic glufosinate resistant plants metabolise glufosinate to NAG. It is also true that a small amount of NAG can be converted into glufosinate on passage through mammalian intestinal tracts. However, the rest of the steps required for Smith's "unique risk" do not occur. NAG appears only at low concentrations, if at all, in grain from glufosinate-treated crops and not at all in processed foods. Therefore, it would be exceptionally difficult to ingest sufficient NAG to even reach the ADI. The vast majority of the NAG and any glufosinate produced from it are excreted rapidly in faeces. Therefore, the chances of consuming sufficient NAG to convert to sufficient glufosinate in the gut to produce any measurable effect must be exceptionally remote."

Petition for approval of LLRICE601 for human consumption [55]

The USDA announced in August 2006 that a scientific review of available data revealed no human health, food safety or environmental concerns were associated with GMO LL Rice 601 (LibertyLink)

In August 2006 Bayer petitioned for the deregulation (approval) for human consumption of LL Rice 601. The USDA has said that the petition is in accordance with its Animal and Plant Health Inspection Service (APHIS) regulations concerning the introduction of GE organisms and products.

Bayer CropScience document: Application for an Extension of the Determination of Nonregulated Status for Glufosinate-Tolerant Rice (98-329-01p): Transformation Event LLRICE601 OECD Unique Identifier BCS-OS003-7

[55]
The undersigned submits this petition under 7 CFR 340.6 to request that the Director, Scientific Services, makes a determination that the article should not be regulated under 7 CFR 340.

Bayer CropScience requests a determination from APHIS that rice with glufosinate herbicide tolerance event LLRICE601 and any progeny derived from crosses of this event with traditional rice varieties, and any progeny derived from crosses of this event with transgenic rice varieties that have also received a determination of nonregulated status, no longer be considered regulated articles under 7 CFR Part 340, and that APHIS consider this document as an extension to petition 98-329-01p.

Glufosinate-tolerant rice based upon the transformation event LLRICE601 was produced by the introduction of a chimeric 35S/bar gene construct using Agrobacterium-mediated gene transfer. The rice events described in petition 98-329-01p were transformed by direct gene transfer of a chimeric 35S/bar gene construct. All events produce the same protein, the enzyme phosphinothricin acetyltransferase (PAT), which confers resistance to the herbicide glufosinate.

Agronomic evaluation has demonstrated that there were no morphological, beneficial organism, disease susceptibility or pest susceptibility differences observed when comparing the events to cultivated rice. (Bayer document, citation end)

Fact sheet Maize [56]

Glufosinate herbicide- resistant GMOs use the GOX genes. The trait Glyphosate Oxireductase (GMEsyn) confers tolerance to glyphosate (Roundup) The donor-organism was originally Ochronobactrum anthropi, but now a synthetic construct is being used.

After treatment with N-acetyl glufosinate (NAG) accumulates in genetically modified plants. Glufosinate was generally a minor component of the residue whereas the main component in the forage, silage and fodder was NAG and the main component in the grain, cobs and husks was MPP (FAO, 1998; Stumpf, 1996).

Information on the metabolism of glufosinate-ammonium and NAG (N-acetyl-L-glufosinate) in laboratory rats, lactating goats and laying hens was reported. In summary, most of the administered dose of both compounds is rapidly excreted. NAG may be partially metabolized back to glufosinate.

In rat studies up to 10% deacetylation occurred at a low dose of 3 mg/kg bw as shown by the occurrence of glufosinate in the faeces. The authors concluded however that most of the conversion was caused by bacteria in the colon and rectum although toxicity findings indicate partial bioavailability (FAO, 1998).

Toxicity studies show a LD50 (orally en subcutaneous) of NAG of more than 2.8 g/kg body weight, indicating that NAG is not toxic (Trinks, 1995).

In samples of Bt-11 maize no glufosinate-derived residues are found in any maize processed fraction which are relevant food or feed items. These include flour, starch, grits and oil. Residues are not detectable in crude and refined oil. In ruminant and poultry feeding studies no detectable residues were found in meat, milk or eggs at the dose calculated to represent the highest residues in livestock feed under Good Agricultural Practice and taking into account the potential use of glufosinate herbicide in several tolerant crops (Scientific Committee on Plants, 1998).

Based on the current knowledge it can be concluded that large-scale applications of transgenic herbicide-resistant crops and glyphosate or glufosinate does not have adverse effects on the safety of food and the safety for users (Hin, 2001).


Sequencing rice

The genetic code of rice is very similar to all the other cereals to a certain extent. Many of the shared genes are in similar positions on the respective chromosomes. A given function of a gene in rice is very likely to be located in the corresponding gene in another cereal."

According to Robin Buell of The Institute for Genomic Research (TIGR). These data can be used to develop new varieties of rice that deliver increased yields and grow in harsher conditions. Rice is genetically similar to maize, wheat, barley, rye, sorghum and sugarcane. Understanding rice opens the door to the genomics of these plants.


Transporter IRTI for assimilation of iron

[57] [58] Professor Mary Lou Guerinot of Dartmouth University and colleagues identified the transporter IRTI for assimilation of iron by plants. The identification of genes involved in iron homeostasis were done at the model plant Arabidopsis. The scientists studied the transport and regulation of gene expression by metals. Iron and zinc, for instance, influence plant growth and affect yield volumes.

The use of phosphate fertilizers, which has led to high levels of cadmium contamination in many locations around the world Cadmium, together with other metals such as manganese, zinc, and cobalt are also transported by the IRTI transporter.

Guerinot used DNA shuffling and heterologous expression in yeast to isolate alleles of IRT1, which allow the iron to be transported but not the poisonous cadmium. The new transgenic seedlings presented the same cadmium level as wild type plants which had lost their IRT1 function. The transgenic plants had twice as much iron as the wild type plants, when grown in the presence of cadmium.

This genetic technique applied in rice could overcome iron deficiency in many poor countries which use rice as staple food.

The research continues looking after which transporters affect the distribution of micronutrients within plants hoping to lead to agronomic benefits such as increased seedling vigour, higher crop yields and resistance to disease.

In her commentary on this research on Golden Rice in Science, Dartmouth biologist Mary Lou Guerinot says that engineering plants to improve their nutritional value would be of great benefit to developing countries. [59]

Food Watch Report "Golden liess" - Golden Rice will not improve micronutrients in diet [60]

The Food Watch report highlights incomplete or not available data related to bioavailability, there were no animal feeding trials performed, safety assessment is incomplete; there are no data on stability of the carotinoids during storage conditions; there are almost no data published related to possible formation of undesired contents of the plant; spread of the genetic material to other plants is most probable.

Alternatives to Golden Rice to reduce vitamin A insufficiency

In past ten years efficient strategies were initiated to reduce vitamin A deficiency in developing countries. These programs are low-priced and can be run targeted where needed. Golden rice is by no way without alternatives, says Food Watch report.

Food Watch asks for a new discussion related to Golden Rice. The two digits grants of the field project may give better results if used for alternative actions, says the report.

The Golden Rice story [61]

Golden rice is a variety of rice produced through genetic engineering to biosynthesize beta-carotene, a precursor of pro-vitamin A in the edible parts of rice, the endosperm.

Milling of rice removes the oil-rich aleurone layer of rice grains to avoid rancidity during storage. Essential nutrients, such as provitamin A are lost during this procedure leading to vitamin A deficiency, in at least 26 countries are the Recombinant DNA technology was used to improve its nutritional value in this respect.

Ingo Potrykus of Switzerland and Peter Beyer of Germany developed the Golden Rice between 1992 and 2000, the year of their first publication. Both scientists signed a deal with AstraZeneca, which, together with Monsanto, agreed to develop the rice for "humanitarian" purposes. In 2005 Golden Rice 2 was presented. It produces up to 23 times more beta-carotene than the original variety of golden rice, introducing a maize psy in combination with the Erwinia uredovora carotene desaturase (crtI) used to generate the original Golden Rice. [62]

Genetic engineering of Golden Rice [63]

Two beta-carotene biosynthesis genes were used to build Golden Rice: psy (phytoene synthase) from daffodil (Narcissus pseudonarcissus) and crtl from the soil bacterium Erwinia uredovora The psy and crt1 genes were fixed to an endosperm-specific promoter, so they are only expressed in the endosperm. The exogenous lyc gene generates geranylgeranyl diphosphate. The bacterial crt1 gene catalyses multiple steps in the synthesis of carotenoids with lycopene which is finally transformed in beta-carotene giving the endosperm a yellow colour.

Anti-GMO activists [64]

Vandana Shiva, an Indian anti-GMO activist says that the causes of nutrients deficiencies are poverty and loss of biodiversity in food crops. GM crops will aggravated the situation by obscuring the larger issue of a lack of broad availability of diverse and nutritionally adequate sources of food. According to Dr Shiva to meet the full needs of 750 micrograms of vitamin A from rice, an adult would have to consume 2 kg 272g of rice per day. This implies that one family member would consume the entire family ration of 10 kg. from the PDS in 4 days to meet vitamin A needs through "Golden rice".

Varied diet containing foods rich in beta carotene such as sweet potato, leafy green vegetables and fruit may provide children with sufficient vitamin A, says Friends of the Earth. A statement of UN FAO, signed by 18 African countries, is quoted saying "we think it will destroy the diversity, the local knowledge and the sustainable agricultural systems that our farmers have developed for millennia, and that it will thus undermine our capacity to feed ourselves." [65]

The best way to eliminate vitamin A deficiency and malnutrition is to use existing diversified food crops at farms and in garden. Promoting Golden Rice diverts of a these efforts, and the introduction of golden rice will further compromise them. Golden rice is merely a marketing event. [66]

Genetic modified potatoes [67]

Dr. Arpad Pusztai from the Rowett Institute in Scotland responsible scientist liberated the news that rats being fed with GMO potatoesinjured in growth and in its immune system.
The results were declared as misunderstanding and Dr. Pusztai was sent into retirement.
A snowdrop-gene had been introduced in the genetical material of the potato. This alien gene should keep worms and insects away. Further research carried out by Dr. Pusztai made clear that there was a potential menace to animals and mankind. Damage of internal organs,malfunction of the immune system and alteration of growth and were demonstrated by rats being fed with the genetic modified potato.
At the moment it is not clear from where the toxicity comes. Some scientists find the promoter-gene to be responsible for it.
The same promoter-gene is being used in the Roundup Ready Soybean.
If this proves to be true all food having soybean derivates from genetic modified soybeans should be considered as harmful to health.

French retailer against genetic modified food [67]

The French retailer Carrefour, Paris announces to withdraw all genetic modified foods.About one third of their own trade-marked articles are affected.
Carrefour is not an enemy of genetic technology, but it seems impossible to predict the results of genetic modification on foods for long terms.Therefore the retailer wants to go the safe way avoiding genetic modified products.
According to Carrefour[67] GMO labeling EU directive proves to be insufficient to provide protection or information for the consumer as to many exceptions are allowed.
Seven important European retailer enterprises have founded a consortium against genetic modified foods. This consortium embraces Migros (Swiss), Carrefour (France), Sainsbury(Great Britain, Marks and Spencer, Delhaize Le Lion (Belgium), Esselunga (Italy) and Superquina (Ireland).
The aim of this alliance is to clean own trade marks from genetic modified ingredients. (vwd/1.4.99/12/mi)

GMO-labeling in USA

US-citizens claim for GMO-labeling in a petition which has been given to the Congress as there are no rules from FDA.

European GM regulation criticized [68]

At an IFT conference on global acceptance and sustainability of genetically modified food 26.06.2006, Francis Smith from the Competitive Enterprise Institute in Washington DC criticized the EU GM food regulations.

She said that consumers in the EU are concerned a lot about food, especially over topics that attract a lot of media coverage, Austrians being the most affected by worries. Fourty percent of European women are deeply concerned about food safety meanwhile only 29 percent of European men feel so.

Smith believes that Europeans fear new technology. Americans do not share this feeling, they look at the benefits and have less fear and less distrust.

Because of the European distrust, triggered by food scandals like mad cow and safety of the food supply chain in the 1990s, decision-makers take into account these fears and adopt prevention strategies.

Smith argued that regulations addressing perceptions and fears rather than scientific evidence, increases the perceptions of risk when there is little basis for this, and could lead to mass hysteria.

According to the WTO, the relevant EC scientific committees had recommended approval of all 25 product applications. But for transparently political reasons rather than concerns about consumer health or environmental protection, EU politicians repeatedly refused to sign off on the final approvals.


France will impose a GM ban if serious safety doubts are presented [69]

The President Sarkozy in October 2007 imposed a temporary ban affecting cultivation of GM crops. This ban will end at the start of February but Sarkozy will restore the ban if scientists raise serious doubts about GM crops.

Austria enforced a ban on the import and processing of MON810 and T25 maize in June 1999 expressing concern about the effects on non-target organisms and the development of resistance to toxins by target organisms but failed to present scientific evidences for their claim. In 2005 the WTO ordered the ban to be lifted.

According to French anti-GM lobby leaded by José Bové pollination could cross-contaminate non-GM crops grown in the vicinity. The long-term health effects of GM on humans are not known. Bové is on hunger strike to enforce the GM ban.

The position of the Association Generale de Producteurs de Mais [70]

The Association Generale de Producteurs de Mais (AGPM) claims that the GM is needed because of considerable damage by corn borers in the south west and centre of the country.

According to AGPM there is absolutely no justification for this decision on a "genetic trait" which has been shown to be harmless and has been recognised as such for a long time, which is approved for use in many countries and has just been put back on the market in Germany after sales were suspended.

The AGPM highlights the implementation of a best practice guide for coexistence and traceability of GM and non-GM corn in 2004, and that no one of the plots studied went above the 0.9% limit set by regulations for labelling. [71]

According to Greenpeace in 2005 the European Commission Monsanto's approved MON810 maize under the old EU directive without a comprehensive monitoring plan. Greenpeace says that the new directive 2001/18/EC should to be applied for MON810. [72]

Austrian long-term study found GM maize to affect reproduction in mice. [73]

GM maize NK603xMON810 from Monsanto was found by Juergen Zentek and colleagues 2008 to affect reproduction in mice, but these findings cannot be applied on humans. Female mice that were fed with the genetic modified maize had fewer babies and fewer litters compared with mice fed with non-GM maize.

The study was presented by the Austrian health and food safety agency AGES, by Vienna's University of Veterinary Medicine. The authors of the study call for more research to confirm the effect of GM maize on other animals and on humans.

French bill sets tough penalties protecting GM crops [74]

On the 22.05.2008 the French parliament passed a bill on general principle for the freedom to produce and consume GM organisms as long as there is no harm to public health or to the environment. However, it does not say how the level of risk to public health will be determined. Long-term health risks on GM exposure are still unknown. [75]

According to the bill the nature and location of GM cultivation will be recorded in a national register. The distance between GM crops and conventional crops will be dependent on the type of plant. Environmentalists say the bill is to weak to prevent cross contamination of conventional crops.

The bill sets tough penalties for GM sabotage

Sabotage of the GM plantations will be severely punished with a two year jail term and a fine of EUR 75,000 for destroying GM crops and more severe penalties if the destroyed crops were intended for research.

The new bill will not end the ban on GM crops in France, however, observers say it is a step back towards GM acceptance. France has the highest increase of GM cultivation in Europe.

Marker-assisted selection (MAS) accelerating traditional breeding [76]

Jeremy Rifkin from the Foundation on Economic Trends reports that new technologies are making gene splicing and transgenic crops (GM foods) obsolete. He urges to look at Marker-assisted selection (MAS) as a method to accelerate traditional breeding. Rifkin says that genes associated with desirable traits such as yield or pest resistance can now be identified, and located in other varieties of the food crop or its wild relatives, which are then cross bred to improve the crop. This technology will eventually replace GM food which uses molecular splicing techniques to transfer a gene from an unrelated species.

Most of the transgenic crops introduced into the fields express only two traits - resistance to pests and compatibility with herbicides - and rely on the expression of a single gene. Marker-assisted selection (MAS) technology could overcome this limitation.

Jeremy Rifkin notes that farmers are gradually forced to used seeds and agrochemical producing crops which are not their own any more, but belong to multinational corporations of agro business and food industry. [77]

According to Nicolas Schauer marker-assisted selection (MAS) in conjunction with transcript profiling is a powerful method to identify genes of interest for the rapid identification of agronomically desirable lines. The combination of MAS and metabolite profiling might prove much more effective in the long run for the identification of metabolic traits in crops. [78]

Herbicides at the base of imidazolinon [67]

Genetic modified rapeseed resistant to the herbicide imidazolinon is already on market.
The pharmaceutical enterprise Cyanamid Canada and the University of Saskatchewan of Canada develops imidazolinon tolerant wheat. The used gene had been detected in winter wheat.

Celiac disease [79]

Celiac disease is caused by allergy to gliadin, an aminoacid from gluten of wheat, rye and barley. Gluten is present in flour of theses grains and is therefore present in all farinaceous foods as well as traces in starch of these plants. Starch is an ingredient of a long chain of industrial products.The German Ministry of Research (BMF Bundesforschungsministerium) coordinates researches to eliminate the genes responsible celiac causing components of the gluten.
Unfortunately industry does not help the project as there is no financial profit expected.Monsanto hopes to release a glyphosate tolerant wheat type in 2002.
Imidazolinon resistant sugar beets were already developed and are on test.

Genetics of Celiac disease [80]

According to Fasano celiac disease is associated with specific HLA class II genes known as HLA-DQ2 and HLA-DQ8 located on chromosome 6p21. Approximately 95% of celiac disease patients express HLA-DQ2, and the remaining patients are usually HLA-DQ8 positive. Either one of these genes is necessary for disease development but additional non-HLA genes are also involved. These additional genes were found by the genome-wide association studies.

It is being suggested that the genetic predisposition to CD is primarily linked to the effect of HLA-DQ2/DQ8 on the immune response to gluten peptides, together with other genes influencing the adaptive immune reactions, intestinal permeability, and a general predisposition to autoimmunity.

The author stresses that the HLA-DQ genes are the only genes for which testing is currently recommended. Presence of symptoms alone does not necessarily predict which individuals might have celiac disease. Individuals with potential CD in high risk groups should be submitted to an accurate, two-step strategy for screening consisting of HLA-DQ typing and longitudinal serologic celiac disease-screening only in subjects positive for HLA DQ2 and/or DQ8.

The presence of DQ2 or DQ8 does not confirm the diagnosis. Conversely, the absence of both HLA types has a negative predictive value of over 99virtually excludes the diagnosis of CD.[81]

The genome-wide association study [82]

A genome-wide association study is an approach that involves rapidly scanning markers across the complete sets of DNA, or genomes, of many people to find genetic variations associated with a particular disease. Once new genetic associations are identified, researchers can use the information to develop better strategies to detect, treat and prevent the disease. Such studies are particularly useful in finding genetic variations that contribute to common, complex diseases, such as asthma, cancer, diabetes, heart disease and mental illnesses.

Cultivation of genetic modified seeds in Brazil

In September 1999 the cultivation of genetic modified soybean [83]will start.
Fields in the state of Rio Grande do Sul and the south of Mato Grosso and Goias will be cultivated with Roundup Ready soybeans from Monsanto over the Monsoy enterprise.The seed was released by the Brazilian commission CTNbio (Commissao Técnica Nacional de Bioseguranca).
Consumer organizations and government sectors have made resistance against GMO soybean. The GMO plant is coming over the border from Argentina.
Hoechst Schering has received the authorization to cultivate herbicide tolerant rice in Brazil.
According to CNTBio (Brazilian biotechnology safety commission) the worldwide culture of GM plants is 28 millions hectares
Soybeans are represented with 40% followed by maize, tobacco, cotton, tomatoes and potatoes.
USA has 8,5 millions hectares of GM plantations, China 1,8 millions hectares, Argentina 1,4 millions hectares and Canada 1,3 millions hectares.
Du Pont de Nemours and Co has entered the Brazilian market with Du Pont do Brasil with its head in São Paulo taking over Pioneer Hi-Bred international Inc in march 99.
As Du Pont has overtaken Sementes Dois Marcos the way is open to the production of genetic modified soy bean seeds together with modified wheat seeds.
The first product of Du Pont in Brazil will be a modified maize seed which was modified without gene transfer so there is no formality necessary to release the seed.This maize has a very high content of oil. (vwd/6.4.99/mi)

Transgenic beans in Brazil, scientists warn from careless use of genetic technology

Brazilian plant geneticist of the states agency EMBRAPA introduced the RNA interference (RNAi) in pinto bean (Phaseolus vulgaris). This GM bean is resistant to the golden mosaic virus, and was approved by the Brazilian National Technical Commission on Biosafety (CTNBio) for human consumption, and will be at the grocery stores in 2014. [84]

The technology was first mentioned in a study of Aragão et al 2007, when the RNAi was protein was described. [85]

According to Francisco Aragão, leading author of the study, small farmers will now profit from the GM pinto bean. Brazil already became the world's second-largest producer of genetically modified (GM) crops, behind the United States, because of non restrictive and funding politics of bot nations. GM maize, soya and cotton are being accepted by the population. Introducing GM beans will increase the exposure of the population to new genes of unknown health risks, because beans are a staple food of Brazil.

Key aspects of the genetic engineering of GM pinto bean are maintained secret

Environmental groups and a presidential advisory panel, the National Council for Food Security and Nutrition, have called for more transparency in biotechnology science and decision-making, and increased research to rule out health risks stemming from the bean. Nodari, a former member of CTNBio complains that key aspects of the genetic engineering were not made public. Edilson Paiva, president of CTNBio, said that Nodari demands that scientists provide the impossible: guarantees of absolute safety, and secrecy is being maintained to allow the patent of the work.

The pinto bean safety controversy [84]

According to Aragão there is no safety concern related to pinto beans because no unfamiliar proteins are produced, only small snippets of RNA, which react with and neutralize RNA from any invading virus are present. Similar RNA snippets are being naturally produced by plants reacting to viral attacks and do no harm to humans argues Dr. Herve Vanderschuren, a Swiss biotechnologist. However, Dr. Jack Heinemann of Canterbury University of New Zealand, commenting the article in Nature, says that small RNA molecules are in our diets, but this does not say that all are safe. It is known that small RNA molecules that cause RNAi have off-target effects. "We know that small RNAs transmit through food to animals. Plant miRNA has now been found in the human circulatory system, probably from food. These miRNAs can effect gene silencing in the mouse liver, and in human cells. Novel small RNAs that are created through this process ought to be checked for having unintended adverse effects on people and key insects and wildlife," he write in the comment. Dr. Heinimann asks for the data of the testing of the pinto bean, so safety concerns could be dissipated.

EMBRAPA starts field trials to determine the yield of the transgenic bean which may then be planted on land on which the golden mosaic virus prevents farmers to grow beans. Aragão predicts a production increase by 10–20%, needed to avoid costly imports of beans. EMBRAPA will also develop the black bean and any other variety to become virus-resistant.

Applying the genetic technology to African cassava

Dr. Herve Vanderschuren plans to apply the same genetic technology to cassava and other local crops in Kenya, Tanzania and South Africa

Vanderschuren et al 2011 developed a method for detection and quantitation of viruses associated with cassava brown streak disease (CBSD), using quantitative RT-PCR (RT-qPCR). The expression patterns of PP2A, UBQ10 and GTPb were found to be host reference genes with stable expression patterns in different Cassava brown streak virus (CBSV)-infected cassava varieties. These reference genes can be used as internal controls for all types of cassava samples as well as in different cassava varieties infected by CBSV. [86]

Micro RNA genes and plant response to stress situation [87]

MicroRNA (miRNA) are short ribonucleic acid (RNA) molecules which have very few nucleotides (an average of 22). MiRNA genes are located in the intergenic regions of the plant genome. They regulate the post-transcriptional expression of proteins in plants and animals. MiRNAs are likely to be involved in most biological processes.

Aberrant expression of miRNAs has been implicated in numerous disease states, such cancer, heart disease and diseases of the nervous system. They are also linked to plant response to biotic and abiotic stress. MiRNA genes and their target genes maintain a gene regulatory network in plants.


Micro RNAs in Cassava [88]

Patanun et al. 2012 studied the miRNAs of cassava, identifying 34 miRNA families The mes-miR319b represent the first mirtron demonstrated in cassava. Mirtrons are a type of microRNAs that are located in the introns of the mRNA encoding host genes and were found in Drosophila melanogaster, Caenorhabditis elegans.and mammals. Patanun and colleagues also described elements relevant to drought stress and plant hormone response in the cassava miRNA genes. These findings may lead to the development of improved plant strains.


Cassava bacterial blight defence system of Manihot [89]

Cassava (Manihot esculenta) is a staple crop which stores important quantities of starch in its roots. These roots constitute the main source of calories for more than half a billion people around the world, mainly in tropical regions and is used for bioethanol production, animal feed and industrial raw materials. An important disease is the cassava bacterial blight (CBB), caused by gram-negative bacteria Xanthomonas axonopodis pv. manihotis (Xam), and produces crop losses up to 80-100%.

Perez-Quintero et al. 2012 studied the miRNAs in cassava under infection with Xam bacteria. Infection increased the expression of some miRNA families, including the miRNAs linked to defence reactions by activating auxin-responding factors. The authors also note that the infection activated families of miRNAs involved in copper regulation and families targeting disease resistance genes, previously known in Arabidopsis thaliana. The miRNAs are important defence system of cassava against Xam and more studies in this direction may lead to varieties with higher resistance to the disease,

Good business with GM-free vitamin E, phytosterols and isoflavones

Fenchem claims to have established a solid supplier chain of indigenous soybean grown in the north-east of China. As more and more GMO soybean seed is being planted US, Brazil and elsewhere, GMO-free soybean products are getting in great demand. Fenchem received IP (identity preserved) certification for soy-derived natural vitamin E, phytosterols and isoflavones.


GMO and worldwide famine, an interview with Ian Wilmut

Can genetic techniques solve the problem of famine in the world?
Ian Wilmut, genetic specialist who cloned the sheep "Dolly" said in an interview with the Brazilian magazine veja (4.11.1998,page 14):
"It would be exaggerated to say so. The actual production of foods is more than sufficient to feed the whole planet. The trouble is that the food is unequal distributed. Instead of producing more food we have to improve the storage of already existing food and we have to distribute them in a fair way. It is sad to know that people still die of hunger just because we do not find a political and social model which enables us to distribute food in a way with more justice. The solution of the famine is political and not scientific. What genetics can do is to give a contribution that this comes true."

Human embrionic stem cells

Human embryonic stem cells (hESCs) are cells that are derived from the inner cell mass of blastocyst stage human embryos, are capable of dividing without differentiating for a prolonged period in culture, and are known to develop into cells and tissues of the three primary germ layers. Although hESCs are derived from embryos, such stem cells are not themselves human embryos. [90]

Stem cells are eligible for US taxpayer funding [91]

The U.S. government approved batches of human embryonic stem cells. These stem cells may now be used in federal funded researches. With the Executive Order 13505 restrictions in the use of stem cells imposed by the foregoing administration, were lifted by Obama in March 9, 2009. The Dickey-Wicker amendment (SEC.509) imposed by the Congress is still active. This amendment forbids the use of federal money for stem cells which require destruction of a human embryo. The use of these cells, made by others, however, are eligible for public funding. [92]

To avoid critics of organisations which say it is wrong to destroy human embryos for any reason, the NIH published the NIH Guidelines for Human Stem Cell Research. [90]

Section 509 [93]

SEC.509: (a) None of the funds made available in this Act may be used for-(1) the creation of a human embryo or embryos for research purposes; or (2) research in which a human embryo or embryos are destroyed, discarded, or knowingly subjected to risk of injury or death greater than that allowed for research on fetuses in utero under 45 CFR 46.204(b) and section 498(b) of the Public Health Service Act (42 U.S.C. 289g(b)). (b) For purposes of this section, the term "human embryo or embryos" includes any organism, not protected as a human subject under 45 CFR 46 as of the date of the enactment of this Act, that is derived by fertilization, parthenogenesis, cloning, or any other means from one or more human gametes or human diploid cells.

Cloning of cells from frozen material

Nuclear reprogramming [94]

Nuclear reprogramming is a term used to describe changes in gene activity that are induced experimentally by introducing nuclei into a new cytoplasmic environment.
The transplantation of a somatic cell nucleus to an enucleated egg results in a major reprogramming of gene expression and switch in cell fate. [95]

Tian and colleagues 2007 assessed nuclear reprogramming and the potential applications in agriculture and biomedicine, which were, however, found to be of low efficiency. The authors compared nuclear reprogramming cells to those generated by conventional reproduction. They found aberrant expression patterns of H19 and Igf2r as well as X-linked genes in term cloned calves, and the expression profiles of cloned blastocystst were considerably different from those of their nuclear donor cells.

The authors concluded that cloned embryos undergone nuclear reprogramming by the blastocyst stage. They stress that re-differentiation may result in gene expression aberrancies, and small initial nuclear reprogramming errors may increase during subsequent development.

Somatic cell nuclear reprograming factors present in the cytoplasma [96]

Bui and colleagues 2000 stress that genomic reprogramming factors are located in the nucleus rather than the cytoplasm of oocytes or zygotes, however, methods using nuclear factors have not succeeded in producing cloned mice from differentiated somatic cell nuclei.

The authors found that there are functional reprogramming factors present in the cytoplasm of germinal vesicle stage (GV) oocytes which could remodel somatic cell nuclei. Bui and colleagues concluded that genomic reprogramming factors are present in the cytoplasm of the GV oocyte and could be useful in cloning technology.

Nuclear reprogramming by somatic cell nuclear transfer timing [97]

According to Zuccotti experiments in 2000 have demonstrated that mouse oocyte can remodel the genetic programme of a somatic cells nucleus. The activation of zygotic genes occurs in the mouse by the 2-cell stage and it is a crucial event in the life of the newly formed mouse embryo as lack or wrong timing of zygotic gene expression leads to the death of the embryo.

The authors concluded that the genetic reprogramming due to the injected somatic nucleus must be completed before zygotic genome activation occurs. Better understanding of the process of cell differentiation and the reversible process, would allow reprogramming without the use of the female gamete.

Somatic cell nuclear transfer cloning [98]

Niemann and colleagues point out thatsomatic cell nuclear transfer (SCNT) is frequently associated with pathological changes in the foetal and placental phenotype and has significant consequences for development both before and after birth. It involves an erasure of the gene expression program of the respective donor cell and the establishment of the new sequence of expression. Implicated processes like DNA methylation, imprinting, X-chromosome inactivation and telomere length restoration in bovine development are discussed by the authors.

Cloned plants differ from their founder cells [99]

Cloning in biotechnology refers to processes used to create copies of DNA fragments (molecular cloning), cells (cell cloning), or organisms. Variations in the resulting unit may occur despite the fact that they are derived from genetically identical founder cells.

Jiang et al. 2011 report that animal clones present genetically variations caused by alteration of the epigenetic reprogramming of their genome. These variations are attributed to the effect of environmental factors on how animal genes are expressed. The authors found that the causes of variation in cloned plants may have different causes and are more frequent compared with variations found in cloned animals. The high number of variations in regenerant plants are due to high mutations in the DNA sequence of cloned cells during the regeneration process itself or during the cell divisions in the donor plan.

Demonstration of the technique of nuclear transfer into mouse oocytes [100]

Dieter Egli and Kevin Eggan explain the technique of nuclear transfer into unfertilized mouse oocytes. This demonstrates that the processes underlying development, differentiation and aging are epigenetic rather than genetic processes.

An epigenetic process is the attaching of a methyl group to a gene, altering or silencing its function, but does not change the gene itself.
The authors stress the possible derivation of stem cells from a patient to replace damaged or missing cells. The authors present a movie on the method of of nuclear transfer developed by the group of Prof. Yanagimachi (WAKAYAMA et al. 1998).

See the movie: Nuclear Transfer, Dieter Egli and Kevin Eggan, Harvard University at: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2504452

The position of the German Ethics Council on human-animal composite beings [101]

The German Ethics Councildraws a very clear separation between humans and animals, even though both belonging to the animal kingdom. The Council cites as moral and law, which are fundamentals of religion and culture. Genetic techniques, developing rapidly since 1980, introduce human genes in mice tailored as model organisms. Nerve precursors won by human stem cells are being implanted in the mouse brain and primates to study Alzheimer's Disease and Parkinson. The Council points to the fact that the brain is a central point to distinct between humans and animals. The question rises on how to react when suddenly such a model organism reveals human cognitive abilities?

The council accepts some experiments on mouse, plant and bacteria, such as Escherichia coli, but rises concerns when human nucleus is implanted in a cows stem cell, whose animal nucleus was removed, such as allowed in UK. Can such human/animal composite beings created by scientist brought to mature beings?

Following such thoughts, the German Ethics Council recommends that certain practices be forbidden: introducing animal material into the human germline, experiments that would lead to the development of human sperm or eggs in an animal, and implanting an animal embryo into a human. The Report of the Council gives therefore much attention to a borderline between humans and animal which is being watered-down by the allowing human-animal- composite beings.

Laws and regulations governing genetic issues in Germany [102]

The German Basic Law (Grundgesetz) [103], in an all-embracing way, refers to the inviolability of human dignity. These wordings embrace all issues of human-animal composite beings. Special issues are treated in the embryo protection law (Embryonenschutzgesetz) [104], the animal protection act (Tierschutzgesetz) [105], the Genetic Engineering Act (Gentechnikgesetz) [106] and the German drug law (Arzneimittelgesetz) [107]. The transplantation act (Transplantationsgesetz) [108] does not cover human/animal issues as it is only related to human tissues and human organs which arre transplated to another human.

Definition of cybrids: Such cytoplasmic hybrids, called cybrids, are handled in the report as a generic term for living organisms, including their early stages, which held human and animal elements (Genes, chromosomes, cell nucleus, cells, tissues, organs). Some laboratories used this technique to insert human cell nucleus into animal oocyte to create stem cells. The German Ethic Council could not get into an agreement related to such cybrid cells.

The Report does not cover xenotransplantation from animal to humans. Xenotransplantation is the transplantation of living cells, tissues or organs from one species to another.

Allotransplantation is the transplantation of cells, tissues, or organs, sourced from a genetically non-identical member of the same species as the recipient, such as human to human.

According to the WHO Xenotransplantation, animal to human, defined as living cells, tissues or organs of animal origin and human body fluids, cells, tissues or organs that have ex vivo with these living, xenogeneic materials, has the potential to constitute an alternative to material of human origin and bridge the shortfall in human material for transplantation. The WHO cautions, however, that this procedure carries risks, in particular of transmission of known or as yet unrecognized xenogeneic infectious agents from animals to human beings and from recipients of xenogeneic transplants to their contacts and the public at large. [109]

The World Health Assembly Resolution WHA57.18 (2004) urges Member States to ensure effective national regulatory control and surveillance mechanisms before allowing xenogeneic transplantation to take place. [110]

The nasty way to spoil your appetite: Have a stem cell steak for dinner [111]

Dr. Mark Post, professor of physiology at Maastricht University in the Netherlands and his team develop meat products grown from stem cell of cattle. Post says that such meat is more sustainable, the test tube meat could avoid the need for livestock. He aims to produce small strands of meat which can be used for the production of burgers and sausages. Much larger pieces of meat, such as steaks and chops will follow.

According to the researcher stem cell meat may help to solve world hunger, but much work must be done to have the technology commercialised in a near future. They plan to produce one burger in the coming year, with a cost of 250 000 Euros/burger, hoping for financial support to start a real manufacturing process.

Global population is predicted to double around 2050 lab grown meats such as beef, chicken and lamb could become commonplace. Fish fillets have previously been grown in a New York laboratory using cells taken from goldfish muscle tissue, and strips of pork were grown in 2009 at the University of Maastrcht.

Cloned animals on your dish [112]

European consumers are less accepting of cloned animal products than their American counterparts, says a research from Kansas State University, USA.

This study says that a significant number of people do have concerns about cloning from an ethical and moral perspective. If these products are labelled as such, a significant number of people will avoid them. USA does not demand labelling for cloned meat.

Students in Europe and in USA were surveyed. American students were found to accept cloned products better than European students. In this study European students were more concerned about cloning from an ethical and moral perspective, and American students were concerned about food safety as the main reason to avoid cloned meats.

The Human Epignome Project must reconsider its standard analytical method [113]

Kriaucionis and Heintz 2009 identified an additional character in the mammalian DNA code, the nucleotide as 5-hydroxymethyl-2'-deoxycytidine (hmdC) which is a constituent of nuclear DNA that is enriched in the brain. The authors suggest that hmdC excerts a role in epigenetic control of neuronal function.

Definition of epigenetics [114]

Epigenetics is the study of nongenetic factors that manage the regulation of gene expression. Shilatifard and colleagues 2009, proposed a definition of in which "an epigenetic trait is a stably inherited phenotype resulting from changes in a chromosome without alterations in the DNA sequence." The authors also proposed three categories of signals establishing a stably heritable epigenetic state: The first is a signal from the environment, the second is a responding signal in the cell that specifies the affected chromosomal location, and the third is a sustaining signal that perpetuates the chromatin change in subsequent generations. The amino acids adenine, thymine, guanine and cytosine (A, T, G and C) make up the DNA code. Epigenetic studies found that two other nucleotide, 5-methylcytosine (5-mC), and the 5-hydroxymethylcytosine regulate the gene expression. Gene silencing takes place when cytosine is methylated to 5-methylcytosine, induced by the enzyme DNA methyltransferase at sites where cytosine precedes guanine in the DNA code. DNA methylation influences the differentiation of embryonic stem cells into the different cell types that constitute the diverse tissues of the body in the embrionic stage. Environmental factors and experiences may cause methylation patterns influencing behaviours that are heritable for several generations.

Standard analytical method to DNA methylation site identification [113]

The identification of the sites of DNA methylation uses bisulfite sequencing. Kriaucionis and colleagues found that this analytical method cannot distinguish between 5-hydroxymethylcytosine and 5-methylcytosine and early studies must be revised, such as the Human Epigenome Project which maps all of the sites of methylation using bisulfite sequencing.

The role of 5-hydroxymethylcytosine is not clear, but the authors suggest that it may play a role in demethylating DNA and have a positive role in regulating gene expression as well.

RGS14 gene limits learning and memory in Mice [115]

According to Hepler and colleagues 2010 learning and memory are linked to strengthening of synaptic connections between neurons of the brain. The authors found that the RGS14 protein can regulate several molecules involved in processing different types of signals in the brain that are known to be important for learning and memory. This protein may control these signals. Deleting the RGS14 gene makes mice smarter with better learning and memory by unlocking the CA2 part of the hippothalamus, a region of the brain whose function is only sparely known.

After new memory formation a long-term potentiation, a strengthening of connections between neurons that can be seen in some parts of the brain, but not in the CA2 region which is resistant to long-term potentiation. Mice with a disabled RGS14 gene, the CA2 region present a strong long-term potentiation.

The authors stress that silencing the RGS14 gene did not cause adverse effects, however, it is possible that hidden brain function changes may not be found yet. Hepler and his colleagues hope to find a compound that inhibits RGS14 or shuts it down to enhance cognition.

Enzyme which converts 5mC to hmC [116]

DNA cytosine methylation is crucial for retrotransposon silencing and mammalian development Rao et al 2009 identified proteins of the TET genes as the 2-oxoglutarate (2OG)- and Fe(II)-dependent enzyme that catalyzes conversion of 5mC to 5-hydroxymethylcytosine (hmC). TET proteins have potential roles in epigenetic regulation through modification of 5mC to hmC. This enzyme may explain the dynamic system of the expression of silencing of genes.

Genetic marker to detect genetic alterations during nuclear transfer [117]

Studying the nuclear transfer (NT) techniques to generate embryonic stem (ES) cells, Wakayama and colleagues 2008 found that cloning is possible using adult somatic cells or embryonic stem cells, and nuclear transfer- embrionic stem cell lines (NT-ES) may be established.

The authors note, however, that embrionic stem cells derived from parthenogenetic embryos show epigenetic disorders.

Parthenogenesis is an asexual form of reproduction found in females where growth and development of embryos or seeds occurs without fertilization by a male. The offspring produced by parthenogenesis almost always are female. Parthenogenesis occurs naturally in some species, including most lower plants, a Kalanchoe succulent plant genus of South Africa, invertebrates (e.g. water fleas, aphids, some bees, some scorpion species, and parasitic wasps), and vertebrates (e.g. some reptiles, fish, and, very rarely, birds and sharks. This differs from hermaphroditic species which can reproduce by themselves because they contain reproductive organs of both sexes. [118]

The researchers conclude that nuclear transfer affects the epigenetic status of a few gene regions and suggest to use the change in the methylation status of U2af1-rs1 as a genetic marker to investigate the effects of nuclear transfer.

Reprogramming skin cells may provide new therapies for liver disesase [119]

Ducan and colleagues 23009 report the reprogramming of mouse skin cells which resemble embryonic stem cell and develop to liver-like cells. These cells may perform liver functions and can be integrated in the liver tissue.

This could lead to the development of therapies which could replace liver transplantation in advanced liver diseases, and be useful in the treatment of diabetes, hyperlipidemia, and hypercholesterolemia.

The research was based on the work of James Thomson and colleagues which described the reprogramming of skin cells, similar to embryonic stem cells, the so called pluripotent stem cells which can develop to any of the body different tissue cells following processes through which liver cells or other body tissues are made during embryonic development. Thomson and colleagues 2009 describe the reprogramming of human differentiated human cells to induced pluripotent stem cells (iPS) using nonintegrating episomal vectors. After removal of the episome, iPS cells are derived that are similar to human embryonic stem (ES) cells. The authors stress that, using this method, the genomic integration or the continued presence of exogenous reprogramming factors are not needed, opening the way to reprogramming human somatic cells for clinical applications. [120]

The use of embrionic stem cells is limited by ethical considerations and by the fact that they do not have the same genetic make-up as the patient. These difficulties could be eliminated reprogramming skin cells of the patient to be treated.

Defining enhancers ruling cell-type-specific gene expression [121]

Heintzman and colleagues 2009 stress that pluripotent embryonic stem (ES) cell identity is governed by a set of transcription factors Oct4, Sox2 and Nanog, and additional transcriptional modulators that bind to chromatin near sites occupied by Oct4, Sox2 and Nanog. This combinatorial control of gene transcription is fundamental to ES cell state, according to the authors.

The variety of functions of the cell of different tissues depends on promoters, enhancers, insulators and other cis-regulatory DNA sequences for each gene. The authors studied their cell-type-specific gene expression role and found that the chromatin state at promoters and CTCF-binding at insulators is largely invariant across diverse cell types, but enhancers are marked with highly cell-type-specific histone modification patterns. The authors defined over 55,000 of such transcriptional enhancers in the human genome explaining their role in cell-type-specific gene expression.

Human induced pluripotent stem (iPS) cells for cardiovascular therapy [122]

Zhang and colleagues 2009 studied the cardiac differentiation potential of human iPS cells generated using OCT4, SOX2, NANOG, and LIN28 transgenes compared to human embryonic stem (ES) cells using the embryoid body (EB) method. The authors found that iPS cells have a capacity like ES cells for differentiation into specific cardiac tissue. They concluded that human iPS cells can differentiate into functional cardiac cells and are a cell source for cardiac repair.

Cloning a 16 years frozen mouse has succeeded [123]

Wakayama and colleagues 2008 report a successful cloning animals by nuclear transfer from bodies kept frozen at -20° for up to 16 years without any cryoprotection. The authors used brain cell nuclei and established embryonic stem cell lines from the cloned embryos, leading to healthy cloned mice.

The authors concluded that nuclear transfer techniques might clone frozen extinct species such as the woolly mammoth frozen extinct species.

Cloning of animals for better foods

[124] The European Commission requested the European Food Safety Authority to advise on food safety, animal health, animal welfare and environment implication of live cloned animals, obtained through somatic cell nucleus transfer (SCNT) technique, their offspring and of the products obtained from those animals. [125]

At present in Europe cloning is not a commercial practice and there is no specific regulation on the authorisation of food products from cloned animals for human consumption in the EU. EFSA's opinion will therefore help inform any future EU measures for cloned animals and their products.

Food safety officials from the 27 member states decided that milk and meat from cloned animals and their offspring should be considered in the same way as any other novel food, such as genetically-modified organisms (GMOs).

To prepare the advice, the EFSA refers to the Opinion Nr. 9 - 28.February 1997 - Ethical aspects of cloning techniques.Opinion requested by the European Commission on 28 February 1997 [124]. This opinion was prepared by European Group on Ethics in Science and New Technologies . [126]

The Group defines cloning as the process of producing "genetically identical" organisms. It may involve division of a single embryo, in which case both the nuclear genes and the small number of mitochondrial genes would be "identical", or it may involve nuclear transfer, in which case only the nuclear genes would be "identical".

But genes may be mutated or lost during the development of the individual: the gene set may be identical but it is unlikely that the genes themselves would ever be totally identical. In the present context, we use the term "genetically identical" to mean "sharing the same nuclear gene set".

According to the European Group on Ethics in Science and New Technologies [126], in their Opinion nr. 9 - 28/05/1997 - Ethical aspects of cloning techniques express that these new technologies increase the power of people over nature and thus increase their responsibilities and duties. The Group cites the potential uses of cloning animals:
- in the field of medicine and medical research, to improve genetic and physiological knowledge, to make models for human diseases, to produce at lower cost proteins like milk proteins to be used for therapeutic aims, to provide source of organs or tissues for xenotransplantation;

- in agriculture and agronomical research, to improve the selection of animals or to reproduce animals having specific qualities (longevity, resistance,...) either innate, or acquired by transgenesis.

From the point of view of animal breeding, the technology could be useful, in particular if it increases the medical and agricultural benefits expected from transgenesis (genetic modification of animals).

By using genetic modification and selection in cultured cell lines, rather than in adult animals, it could become possible to remove genes, such as those provoking allergic reactions, as well as adding genes, for the benefit of human health.

Strong public concern related to animal cloning for food [127]

The Eurobarometer survey on public attitutes toward animal cloning revealed a strong public concern related to cloning for food production such as meat and milk. Animal cloning has already been introduced in the US food industry and the European Commission expects that meat and milk from cloned animals will be worldwide on the shelves of retailers by 2010.

The European Commission will decide on cloning based on the results of the Eurobarometer survey and a report of the EFSA on the safety of foods from cloned animals.

The Eurobarometer survey on consumers' perception of animal cloning, main outcomes

Animal cloning is the creation of a genetically identical copy of an existing or previously existing animal. The Flash Eurobarometer survey asked citizens of the EU about their perception on cloning: to clarify their attitudes towards animal cloning, and its perceived effects on a number of areas including food safety, ethical and animal welfare concerns.

A large majority of EU citizens (81%) said they new the meaning of animal cloning.

The vast majority of EU citizens agreed that the long-term effects of animal cloning on nature were unknown, animal cloning might lead to human cloning, animal cloning was morally wrong, and cloning might decrease the genetic diversity within livestock populations.

The majority of EU citizens would not accept animal cloning for food production purposes, food industry would ultimately be benefited, only a quarter of the population trust.

25% of the interviewees selected scientists as the most trusted source for information. about the safety of cloned animals.

A majority of EU citizens said that it was unlikely that they would buy meat or milk from cloned animals, even if a trusted source stated that such products were safe to eat.

Eight out of 10 EU citizens said that special labelling should be required if food products from the offspring of cloned animals become available in the shops.

Opinion of the European Food Safety Agency [128]

The European Food Safety Authority (EFSA) is the keystone of European Union (EU) risk assessment regarding food and feed safety. The EFSA in its opinion on the safety of foods from cloned animals stated: "There is no indication that differences exist in terms of food safety for meat and milk of clones and their progeny compared with those from conventionally bred animals. However, such a conclusion is based on the assumption that meat and milk are derived from healthy animals which are subject to relevant food safety regulations and controls. Only pigs and cattle are addressed in this opinion: the two species of animals where adequate data were available." The agency, however, stressed that data were limited, samples were of small size and log time observation was not available.

The EGE opinion [129]

The The European Group on Ethics in Science and New Technologies to the European Commission (EGE) said in a Report in January 2008 that it did not see convincing arguments to justify the production of food from clones and their offspring.

Considering the current level of of animal clones, the EGE find it ethically not justified to clone animals for food supply considering suffering and health problems of the animals. The EGE says that it does not see convincing arguments to justify the production of food from clones and their offspring.

The EGE explains that the Somatic Cell Nuclear Technique (SCNT) used for animal cloning is via three main steps:
  1. enucleation of an oocyte, performed mechanically by fixing the oocyte and aspirating the nucleus by using a sharp glass pipette.
  2. transfer of the nucleus from the donor cell into the fertilized enucleated oocyte by using a micro-manipulator and a microscope and applying a short electrical impulse.
  3. activation and reprogramming of the reconstructed embryo. The exact mechanisms are still unknown.
When the cloned embryo resulting from SCNT starts to develop it is transferred to a surrogate mother, which carries out the pregnancy. So far, around a dozen animal species have been cloned via SCNT. Animal clones that are relevant for the food market include sheep, goats, bovines and pigs. According to the EGE report 20% of cloned calves die in the first 24 hours of birth and a further 15% before weaning. Those that survive the early stages are likely to suffer in later in life through high mortality rates, ill health and susceptibility to multiple organ failure. Cloned foetuses are also often 25% heavier than normal which causes severe problems for surrogate mothers when giving birth.

MEP Intergroup on Animal Welfare

[130]
According to Neil Parish, chairman of the Agriculture Committee, and Member of the European Parliament, the current animal cloning technology is considered primitive and many clones die in the early stages of life. Parish concludes that cloned animals suffer from many more ailments and generally live far shorter lives. From an agricultural perspective, there are serious questions over the effect of this on the gene pool, making cloned animals far more susceptible to disease.

The European Commission should move to ban animal cloning for food until the science has proven it is safe for them to enter the food chain.

US Center for Food Safety [131]

According to the Center for Food Safety the Food and Drug Administration's (FDA) in January 2008 determined that milk and meat from cloned animals are safe for sale to the public. In addition, the FDA is requiring no tracking system for clones or labelling of products produced from clones or their offspring.

In its risk assessment of cloned food, the FDA claims to have evaluated extensive peer reviewed safety studies to support its conclusion, yet a recent report issued by CFS shows the assessment only references three peer-reviewed food safety studies, all of which focus on the narrow issue of milk from cloned cows. [132]

Recent opinion polls show the majority of Americans do not want milk or meat from cloned animals in their food.

The Centre for Food Safety welcomes the Mikulski-Specter amendment of the Farm Bill addresses the gaps and inadequacies of the FDA's current risk assessment. The amendment requires a rigorous and careful review of the human health impacts of allowing clones food into America's food supply to be conducted by the National Academy of Science before any food products from clones are marketed. It also directs the USDA to examine consumer acceptance of cloned foods and their likely impact on domestic and international markets. [133]

Meat and dairy products from cloned animals in the EU

Opinion of the EFSA on cloned foods [134]

The European Food Safety Authority (EFSA) concluded there is no expectation that clones or their progeny would introduce any new food safety risks compared with conventionally bred animals. According to the EFSA meat and milk obtained from healthy cattle and pig clones and their offspring are within the normal range with respect to the composition and nutritional value of similar products obtained from conventionally bred animals.

The opinion, however, says that there are only limited data on safety of cloned food and information on the survival of cloned animals. The EFSA opinion may influence the approval of cloned foods in USA.

The EFSA launched a public consultation on the draft scientific opinion on the implications of animal cloning on food safety, animal health and welfare and the environment. [135]

Cloning makes multiple exact copies of a single gene or other segment of DNA. The animals thus produced have the same genetic material as the original animal, allowing to create strains of animals with increased disease resistance and other qualities.

There are no food safety concerns about cloned foods, but animal health and welfare is compromised says EFSA [136]

In its final scientific opinion on the implications of animal cloning on food safety, animal health and welfare and the environment the EFSA stated in June 2008. Some of the key conclusions of the opinion include:

-There are significant animal health and welfare issues for surrogate mothers and clones that can be more frequent and severe than for conventionally bred animals.

- There are only little data available concerning cloned foods.
- A high number of cattle and pigs clones presented severe health and welfare adverse anomalies.
- Somatic Cell Nucleus Transfer (the most common technique used to clone animals) has resulted in the production of healthy cattle and pig clones.
- No differences between food products from clones or their offspring, in terms of food safety, compared to products from conventionally bred animals if healthy animals which comply with the relevant food safety regulations and controls.
- No environmental impact is foreseen but there are only limited data available.
- The health and welfare of clones should be monitored during their production life and natural life span, and further causes of pathologies and mortality, the immunocompetence and the susceptibility of clones and their offspring to diseases and transmissible agents should be investigated.

Offspring of a cloned cow put to sail on march 2008 [137]

The BBC News report (05.03.2008) that a commercial breeder placed two offspring of a cloned cow to sail on an auction at Easter Comton UK. However, the animals were withdrawn later on because of negative publicity.

The Compassion in World Farming (CIWF) group said that the selling would bring cloning out of the laboratory and on to the farm. The group stresses that they are opposed to cloning on the basis of the animals' welfare.and also there has not been enough research into whether or not there are dangers of the meat or milk entering the food chain.

Coldiretti Italian farmers' group

The Italian farmer' group Coldiretti says that cloned foods are an unacceptable risk. [138]

FDA wants to introduce cloned foods

According to the World Society for Animal Protection cloning results in animal suffering. with 95% of cloning attempts failing. In addition, many of the animals being cloned are those with particularly high productivity, such as cows with excessively large udders that produce more milk but also suffer from major leg problems and painful diseases. [139]

The agency is not requiring labeling or any other additional measures for food from cattle, swine, and goat clones, or their offspring because food derived from these sources is no different from food derived from conventionally bred animals, however, there are insufficient information of other animal species, such as sheep, which are recommended not to enter the food chain. [140]

Should a producer express a desire for voluntary labeling (e.g., "this product is clone-free"), it will be considered on a case-by-case basis to ensure compliance with statutory requirements that labeling be truthful and not misleading. The Agengy says that it did not consider moral, religious and ethical concerns aspects of cloning. The risk assessment was strictly a science-based evaluation. [140]

The Mikulski and Specter amendment [141]

Amendment 3524, introduced by Senators Mikulski and Specter, calls for more information on food products from cloned animals, with specific focus on elements that have not been addressed by the Food and Drug Administration's (FDA) initial risk assessment and reassures the moratorium on cloned foods.

The Food and Drug Administration FDA's preliminary report in December concerning the safety of cloned foods was criticized for being flawed and incomplete. According to the Center for Food Safety (CFS) there are few studies on the risks of food from clones, and no long-term food safety studies have been done. FDA indicates that it will not require labeling on cloned food, so consumers will have no way to avoid these experimental foods. The US Food and Drug Administration is expectedto give its final verdict on food from cloned cattle and pigs soon. [142]

The proposed amendment to the Farm Bill calls for studies that would evaluate the health effects of allowing the commercialization of milk and meat from cloned animals.

CSPI says that if companies begin using clones to breed food animals, they need to explain why. Will it make any food product better, safer, cheaper or more sustainable? Clear evidence of benefits must be generated if consumers are going to accept cloned animals and their products. [143]

Cloned meat in USA

The US Food and Drug Administration (FDA) said it planned to approve cloning for food production in 2007 and to allow the product into the food chain without the need for labelling.

Cloned meat and milk already entered US food chain [144]

According to Washington Post the U.S. cattle cloning companies did not tracked how many offspring of clones have entered the food supply. Producer say that cloned Kansas cattle semen were openly sold to many U.S. meat producers in the past years.

USDA encourages a voluntary moratorium [145]

USDA says that currently, the cloning industry is adhering to a voluntary moratorium on putting cloned products into the food chain. The Agency encourages the cloning industry to continue its voluntary moratorium for a sufficient period of time to prepare so that a smooth and seamless transition into the marketplace can occur.

Cloning ethically not justified [146]

The European Group on Ethics in Science and New Technologies (EGE) has doubts as to whether cloning animals for food supply is ethically justified. Whether this applies also to progeny is open to further scientific research. The organisation does not see convincing arguments to justify the production of food from clones and their offspring, and recommends in case of future food products derived from cloned animals introduction to the European market, that the following requirements are met:

Food safety is considered to be a pre-requisite for the marketing of foods for human consumption. Concerning animal welfare and health, the five freedoms, should be met in intensive animal breeding: from hunger, thirst and malnutrition; from fear and distress; from physical and thermal discomfort; from pain, injury and disease;and to express normal patterns of behaviour should be maintained. A Code of Conduct on responsible farm animal breeding, including animal cloning should be prepared. Current EU legislation regarding traceability of animals and their food products should be enforced.

Despite the efforts of open markets of the WTO, the import of cloned animals, their offspring and materials derived from cloned animals (e.g. semen and food products) should be conditional on proper documentation, in particular with regard to traceability provisions and animal welfare and initiatives to ensure consumers' freedom and rights should be taken. Proper measures to preserve the genetic heritage of farm animal species should be installed.

Public discussion related to the use of animal cloning should be promoted and indicators on public perception concerning the introduction of such products to the food market should be collected. Labelling procedures should be cleared prior to the marketing of such food in the EU.

The Food Standards Agency position [147]

Foods produced from cloned animals fall under Regulation (EC) No 258/97 (Novel Foods Regulation). This means that meat, milk or eggs from cloned animals would be subjected to a safety evaluation and approved by all European Union (EU) Member States as a novel food before they could be legally marketed. In U.S. products from cloned animals and their offspring are allowed by FDA to enter the food chain. This may lead to the technology being considered for use in Europe. [148]

According to UK Food Standards Agency foods like meat milk and eggs from cloned animals are not allowed to enter the food chain in UK. [149]

There are no methods to detect offspring of cloned animals. As sperms from these animals are already being sent all over the world an unknown safety issue will affect the whole food industry with farming Frankenstein creatures.

Cloning of animals

The Group gives their opinion concerning cloning of animals:
Research on cloning in laboratory and farm animals is likely to add to our understanding of biological processes, in particular ageing and cell commitment, and hence may contribute to human well-being. It is ethically only acceptable if carried out with strict regard to animal welfare, under the supervision of licensing bodies.

Cloning of farm animals may prove to be of medical and agricultural as well as economic benefit. It is acceptable only when the aims and methods are ethically justified and when it is carried out under ethical conditions.

These ethical conditions include:
- the duty to avoid or minimize animal suffering since unjustified or disproportionate suffering is unacceptable;

- the duty of reducing, replacing and when possible refining the experimentation adopted for the use of animals in research;

the lack of better alternatives;

human responsibility for animals, nature and the environment, including biodiversity.

The group stresses the need to preserve genetic diversity in farm animal stocks. Strategies to incorporate cloning into breeding schemes while maintaining diversity should be developed by European institutions.

On regard to human reproductive cloning the Group writes that the European Community should clearly express its condemnation of human reproductive cloning and should take this into account in the relevant texts and regulations.

According to the Group, further efforts must be made to inform the public, to improve public awareness of potential risks and benefits of such technologies, and to foster informed opinion.

Cloning of animals for food should be prohibited says MEPs [150]

The European Union is currently considering whether to approve animal cloning for food production. According to Sonja Van Tichelen , director of the Eurogroup for Animals cloning is an incredibly wasteful way of producing food, and causes suffering and harm to animals at every stage of development.

Cloning has been proved to be an inefficient practice that requires the loss of many animal lives just to produce one successful clone. Scientists have found that the ones who do survive suffer more defects and die much earlier than non-cloned animals. Only 8% of sheep involved in a cloning process result in a viable offspring or embryo transferred. For cows this is 15-20%. Goats less than 3%, pigs 3-5%, rabbits less than 2%, mice less than 2%, horses less than 1%, and deer less than 1%.

Members of the European Parliament of the Intergroup on Animal Welfare, leaded by Neil Parish MEP, signed a resolution calling on the Commission to prohibit:
  1. The cloning of animals for food supply
  2. The farming of cloned animals or their offspring
  3. The placing on the market of meat or dairy products derived from cloned animals or their offspring
  4. The import of cloned animals or their offspring, semen and embryos from cloned animals or their offspring, and meat or dairy products derived from cloned animals or their offspring.

Intellectual property issues

The Ethic Group in its Opinion 23[146] stresses that it should be clarified whether the exclusion clauses in Directive 98/44/EC (Art. 6)[151] on patentability of biological inventions and the EPO rules (23) to animal cloning for food apply.


Patents on GMO-plants

The DBV (Deutscher Bauernverband)German Farmer Union has urged the German Government not to take over the European Biopatent guideline in the German Patent law. The DBV says that European Commission opens the possibility of o wide spreadread patent on plants.It is also possible that patent protected gens spread in nature contaminating other plants Farmers would be than unwillingly colliding with patent claims. According to the German Farmer Union are plants and animals a part of natural life basics and should not be withdraw from general availability. Plants and animals are part of nature and should not be put under the monopoly of certain business groups.

Assisted reproductive techniques (ARTs) [146]

Assisted reproductive techniques (ARTs)are:
Image clone


Animal cloning using somatic cell nuclear transfer (SCNT)

Whereas all of the current methods still involve taking sperm from a male and joining it with an egg from a female, cloning is the ability to create an exact copy of a biological entity by means other than the joining of a sperm and an egg. Genetic diversity is strongly reduced in the offsprings. The lack of genetic diversity makes the species vulnerable to environmental changes like new diseases and early aging.


Artificial Insemination

Involves the collection of semen from bulls which can then be used to artificially inseminate cows. It is possible to freeze the semen, store it for longer and make it more widely available. Genetic diversity is not affected.


Oestrus Synchronisation

Involves giving hormone implants and injections to ensure a group of cows are ready for breeding at the same time, being used together with artificial insemination. Genetic diversity is not affected.


Embryo Transfer

Just as artificial insemination is used to produce lots of calves from a single, prize bull, embryo transfer is used to produce more offspring from a prize cow stimulated with a hormone to produce a number of eggs at once. The cow is then mated and many of the eggs are fertilised and start to develop. The embryos are flushed out surgically and implanted into the uterus of surrogate mothers where they are brought to term. Genetic diversity is not affected.


In Vitro Fertilisation

This is a similar process to embryo transfer except the unfertilised eggs of the prize cow are fertilised in an incubator before being transferred to surrogate mothers. Genetic diversity is not affected.


Sexed Semen

The purpose of this is to produce a higher proportion of female calves. Genetic diversity is not affected.


GMO rapeseed in Europe

GMO-rapeseed has been released in Europe by Advanta Seeds without knowledge of the farmer. The GMO-seed was distributed in large scale in Germany,France and Sweden and England for two years. France and Sweden gave order to destroy the GMO- fields. German Environment Ministry says that there is no regulation to force the destruction of the German fields as there is no direct menace to public heath. This is a sign of inexcusable weakness of the German Environment Ministry which is unable to protect purity of nature. Advanta Seeds says that it was a mistake but it is supposed to be a strategy of the seed companies to contaminate rape seeds with GMOs in order to avoid the GMO-free alternative to GMO-Soya which is being refused by retailers in Europe.
The contamination of nature is getting ahead because logistics, flying pollen, handling during processing in storage, on ships and trucks.


GMO Sugar beets in Germany

According to daily news from 10.10.2000 sugar beet plants were genetically modified and not allowed released by KWS SAAT AG in Einbeck (Kreis Northeim) Germany. They are resistant to the herbicide Round up. KWS SAAT AG says that it was a technical mistake which caused the release. This is the proof that genetic technology is being handled careless. It also may be a part of a strategy to contaminate nature in order to avoid the GMO-free alternative. It is possible that the gene passes over to weed turning these plants resistant to herbicides.
To counter the growing contamination of nature with GMOs the European retailers urge the producers to guarantee GMO-free production. Even the 1% contamination which is granted as unavoidable contamination with GMO in connection with GMO-free labeling is not allowed in their products.


Recall of Aventis GMO maize "StarLink"

Aventis has started a recall of its GMO maize "Starlink" according to press release from 12.10.99 due to pressure from US- Agriculture Ministry and the Environment Protection Agency ( EPA ).
StarLink is a variety of Bt corn that has been genetically engineered to produce a protein, Cry9C, intended to be toxic to certain insect pests of corn.

StarLink maize was liberated for animal feed. This GMO maize was sold by farmers to the Azteca Mill in Texas. The maize meal produced from StarLink seed was sold to Kraft as ingredient to the Taco-Bell foods.
Half of the crop of StarLink was sold to the farmers of Iowa. In Europe StarLink was mingled with non-GM seeds. Now almost all plantations are potentially contaminated with StarLink.
Starlink Maize contains a protein which is supposed to cause allergic reactions on humans.
Some Japanese importers denied to import StarLink-Maize as animal feedstuff to avoid the possibility of being used for human food.
Aventis Crop-Science says there will be no sells on seeds of Starlink in 20001.


Monarch butterfly and the Bt maize

Many reports affirmed that the Monarch butterfly was killed by toxic compounds of the Bt maize which has parts of the genes of Bacillus thuringiensis. According to the Environmental Protection Agency (EPA) reviews of scientific informations indicates that there are very little risks for the butterfly. Some authors even predict benefits to the butterfly from farming of corn, cotton and potato Bt plants.
John Obycki and Laura Hansen of the Iowa State University had demonstrated that larvae of monarch butterflies grew more slowly and suffered a higher mortality rate when being fed milkweed leaves with deposition of pollen from insect pest tolerant GM maize as sole food source to monarch larvae, causing 20% mortality in larvae within 48 hours with one variety of GM maize. The Environmental Protection Agency trying to reach decision regarding renewal of registrations for several Bt products argues that the situation is different from that prevalent in natural environment.
The larvae of the monarch butterfly feed on milkweed plants mostly in June whereas the peak time of maize pollen shed is from mid-July to August.This allegation however is insignificant as toxic materials for living beings are present bt maize is therefor harmful for nature.

GM Maize Mon 863

At the request of the European Commission the Scientific Panel on Genetically Modified Organism (GMO Panel) of the European Food Safety Authority (EFSA) has reviewed an evaluation report concerning GM maize MON 863 submitted by Germany regarding the safety of the insect-protected genetically modified maize MON 863. Results of the rodent toxicity study with MON 863 maize did not indicate concerns about its safety for human and animal consumption. (20 October 2004)

Serious concerns regarding the food safety of GM corn MON 863 [152]

The maize for animal and human in many countries, including the EU, Japan, Mexico and the USA.

Signs of liver and kidney disruption: Seralini and colleagues at CRIIGEN (Committee for Independent Research and Genetic Engineering) based at the University of Caen, in a study supported by Greenpeace, found signs of liver and kidney toxicity in rats fed with transgenic maize MON863 which expresses the bt-toxin (Cry3Bb1) protecting against the corn rootworm pest. According to the researchers, It appears that the statistical methods used by Monsanto were not detailed enough to see such disruptions in biochemical parameters.

Difference in weight gain [153]

The authors found significant differences in the weight gains data , with differences between male and females, and suggest that this could be due to endocrine disruption and/or hormonal metabolism differences caused by the GM corn MON863.

Cause of toxicity not known

It is not known whether the signs of toxicity are caused by the Bt protein, or from some changes in the plant's own DNA caused by the genetic engineering event. The authors cannot conclude that GM corn MON863 is a safe product. Companies should be more rigorous in the studies and ensure that their data stands up to scrutiny, and the regulatory authorities such as the EFSA in Europe [154], should demand more complete data before approving any crop.

EFSA reaffirms its risk assessment of genetically modified maize MON 863 [155]

The EFSA has examined a paper by Séralini et al. on the statistical evaluation of a 90-day feeding study in animals with genetically modified maize MON863, to identify any consequences for EFSA's risk assessment of the safety of MON863. The paper presents an alternative statistical analysis of the 90-day rat study that was considered in the original risk assessment.

Following a detailed statistical review and analysis by an EFSA Task Force, EFSA's GMO Panel has concluded that this re-analysis of the data does not raise any new safety concerns.

The main conclusions are:
- The statistical analysis made by the authors of the paper did not take into account certain important statistical considerations. The assumptions underlying the statistical methodology employed by the authors led to misleading results.

- EFSA considers that the paper does not present a sound scientific justification in order to question the safety of MON 863 maize.

- Observed statistically significant differences reported by Monsanto, Séralini et al., and EFSA, were considered not to be biologically relevant. Therefore, the GMO Panel sees no reason to revise its previous Opinions that the MON 863 maize would not have an adverse effect in the context of its proposed use. Prior to this most recent work, MON 863 maize has been subject to a comprehensive risk assessment by EFSA and by other authorities which did not identify any adverse effects on human and animal health or the environment. In addition to the original Opinion in April 2004, this study has been reviewed again twice since then, prior to this recent work.

Other related documents:
Letter to the Commission
http://www.efsa.europa.eu/en/about_efsa/structure/who_is_who/home_cgl/correspondence.html
The GMO Panel statement
http://www.efsa.europa.eu/en/science/gmo/statements0/gmo_statement_mon863_ratfeeding.html
EFSA statistical analysis of the Monsanto data
http://www.efsa.europa.eu/en/science/scientific_reports/statistical_analyses_MON863.html

EFSA Safety and Nutritional Assessment of GM Plants and derived Food and Feed released [156]

The safety assessment of GM plants and derived food and feed follows the approach widely accepted by international organisations such as the FAO/WHO, Codex Alimentarius and the OECD. It is based on comparison with conventional counterparts to identify intended and unintended differences.

The Report is a basis for deciding whether animal feeding trial studies are needed for the safety and nutritional assessment of GM food and feed. The report was published by the EFSA in March 2008.

The EFSA is working with member states to update risk assessment guidance for GMOs. These guidelines remain voluntary, however all member states do use them.

Safety assessment of genetically modified crops [157]

Horizontal gene transfer from genetically modified crops to gut microflora most likely occurs with transgenes of microbial origin. Kleter, Peijnenburg and Aarts reviewed the function and characteristics of microbial transgenes and their horizontal transfer other than antibiotic-resistance genes in market-approved genetically modified crops. The authors concluded that the microbial genes reviewed in their study do not give rise to health concerns, and recommend to include all other transgenes not assessed in this study in the premarket safety assessment of genetically modified crops carrying transgenes.

The authors analyse the speculations that an increased survivability of pathogens in the environment may indirectly increase the likelihood of exposure to these pathogens. The authors associate this with the presence in soil fungi of analogues of the bacterial gene coding for beta-glucuronidase might originate from a horizontal transfer between bacteria and fungi, which have been reported.

The authors stress that safety assessment should follow internationally harmonised approaches such as the FAO/WHO Codex Alimentarius guidelines for the safety assessment of foods derived from GM plants and microorganisms. [157] [158] [159] [160] [161] [162] [163]

The authors point to the fact that the transfer of herbicide resistance genes and others may, in theory, provide a selective advantage to soil microorganisms sensitive to herbicide action, and an increased survivability of pathogens in the environment may indirectly increase the likelihood of exposure to these pathogens.

Assessed transgenes [157]

Bromoxynil nitrilase (BXN)

Genetic engineering of BXN into crop plants renders them resistant to application of the herbicide bromoxynil. No direct impact of this gene on human or animal health is foreseen if it were to be transferred to pathogens given the apparently specific role of this gene in soil environments, according to the authors.

3-enolpyruvylshikimate-5-phosphate synthase (cp4 EPSPS)

The source of the cp4 epsps gene was the soil bacterium Agrobacterium strain CP4, which was one out of a group of glyphosate-degrading bacteria. The authors concluded that there is no indication that the potential transfer of the cp4 epsps gene from GM crops to microorganisms would alter the pathogenicity.

Glyphosate oxidoreductase (GOX)

The source organism Achromobacter LBAA was one of the bacteria isolated from activated industrial and domestic sludge that were capable of degrading glyphosate. Glyphosate, which is converted by GOX, has no role in the treatment of human and animal disease. A potential transfer of GOX from transgenic plants may therefore, not increase pathogenicity of recipient microorganisms.

Phosphinothricin acetyltransferase (PAT)

Streptomyces hygroscopicus (bar gene) and Streptomyces viridochromogenes (pat gene) are streptomycetes that occur in soil and produce the natural herbicide bialaphos (phosphinothricin-alanine-alanine). PAT shows substrate specificity for phosphinothricin and similar compounds, a natural antibiotic which is not used in treatment of human and animal disease. The transfer of the PAT enzyme is unlikely to confer increased pathogenicity to pathogens.

Barnase and Barstar

The genes encoding Barnase and Barstar have been cloned from Bacillus amyloliquefaciens which is not pathogen. The authors concluded provisionally that the potential transfer of the barnase and barstar transgenes is unlikely to influence the pathogenicity of recipient microorganisms.

DNA adenine methylase (DAM)

The dam gene has been isolated from Escherichia coli. Genes encoding adhesins, siderophores, and toxins, are linked with mobile genetic elements are not present in non-pathogenic strains used to derive the dam transgene. Transfer of the dam transgene would not impact on the pathogenicity of recipient microorganisms, according to the authors.

1-aminocyclopropane-1-carboxylate (ACC) deaminase

The gene encoding the 1-aminocyclopropane-1-carboxylate (ACC) deaminase has been isolated from Pseudomonas 6G5. The authors concluded that the potential transfer of the ACC deaminase transgene from GM crops is unlikely to contribute to pathogenicity of recipient microorganisms.

S-adenosylmethionine hydrolase (SAMase)

The SAMase gene encoding the enzyme S-adeno-sylmethionine hydrolase (SAMase), is derived from the E coli bacteriophage T3, which prevents bacteriophages from degradation by infected bacterial hosts. The authors concluded that horizontal transfer of SAMase is unlikely to contribute to pathogenicity to recipient bacteria.

β -glucuronidase (GUS)

The uidA gene, which codes for the beta-glucuronidase (GUS) enzyme, has been isolated from E coli. The authors concluded that the horizontal transfer of GUS will not pose a safety risc.

Nopaline synthase (NOS)

The native tumor-inducing plasmid pTiC58 of Agrobacterium tumefaciens strain C58 harbours among others the gene nos encoding nopaline synthase. The authors say that a horizontal transfer to microorganisms would not affect human and animal safety.

Cry proteins

The source of the cry genes used for genetic modification of crops is Bacillus thuringiensis, which produced spores containing crystals that are toxic to insects. Reviews of safety data found no toxicity of Cry proteins to humans. The authors found no threat to humans and domestic animals should a transfer to microorganism take place.

Nitrogen-fixing bacterium's genome

The complete genetic sequence of the nitrogen fixing bacteria Sinorhizobium meliloti[164] has been published. Nitrogen is essential for the growth of plants. It can be supplied in form of:
1. Nitrogen fertilisers such as ammonium ions (NH4) which is 30% of the total amount needed
2. Atmospheric nitrogen are built through various natural processes representing 40% of the total amount needed by the plants. 3. Atmospheric nitrogen obtained by reduction caused by symbiosis between legumes and bacteria known as Rhizobium, reducing atmospheric nitrogen into ammonium ions which can be used for the synthesis of proteins and nucleic acids.


Endosymbiosis of green algae with embryo of the spotted salamader [165]

Kerney et al 2011 describe the endosymbiosis of green algae within cells of embryos of salamander. Endosymbiosis is vary rare because of the immune reaction of the host cells which try to defeat any foreign cells which manage to get inside of their cells.

The green algae Oophilia amblystoma, living in cells of salamander embryos was described by Lambert Printz in 1927. Later on it was discovered that salamander embryos did not develop completely and quickly in absence of the green algae. According to the authors the alga can invade and grow within an egg's jelly. Once inside, it metabolizes the carbon dioxide produced by the embryo and provides it with oxygen as a result of photosynthesis. [166]

To improve the detection of the green algae cells the authors used fluorescent microscopy to turn chlorophyl of the algae visible together with a short string of nucleic acids that targets and binds to a ribosomal RNA molecule unique to Oophilia (18S rRNA)

Endosymbiosis [167]

The authors explain that symbiosis is a species-species interaction in which the organisms share space for extended periods of time. Symbioses can benefit one organism and harm the other (parasitism), benefit both (mutualism), or benefit one creature and leave the other unaffected (commensalism). Endosymbiosis is a special type of symbiosis, requiring one organism to live inside the cells of another. Endosymbiosis has special evolutionary significance, as it may have preceded the full integration of certain cell organelles, such as mitochondria and chloroplasts.

Australian GM field peas

Field peas in Australia were heavily attacked by the field pea weevil Bruchus pisorum. The Commonwealth Scientific and Industrial Research Organisation (CSIRO) developed a GM field pea which was resistant to the weevil.

Alfa-amylase blocking gene of GM field peas

A gene from beans introduced in the field pea blocked the activity of the alfa-amylase enzyme. This enzyme is important for digestion of starch. Weevil larvae feeding on starch in the developing pea seed are unable to digest the starch and starve.

Beans with the alfa-amylase inhibitor gene were eaten by humans for many years without evidence of an immune response. However, according to Dr. Simon Hogan and Professor Paul Foster at the John Curtin School of Medical Researche the peas with the same alfa-amylase inhibitor gene did cause an immune response in mice, resulting in inflammation of the lung tissue .

Subtle changes that occurred in the chemical structure of the bean alfa-amylase inhibitor as it was introduced in the field pea are responsible for the immune responses. These changes in structure is probably being caused by glycosylation, which occurs when proteins are made via a particular pathway in cells.

The introduced bean protein had to go through a pathway in cells where it would undergo several processing steps including glycosylation. Glycosylation can differ from organism to organism and even in different cell types within an organism causing the immune response in GM field peas.

CSRIO decided not to progress development of these GM field peas. [168]

Lycopene from Fusarium sporotrichoides

Lycopene is the red colour of tomato and is used in nutraceuticals as strong antioxidant, reducing risk of prostate cancer and heart diseases. It is obtained by extraction and purification from the carotinoids from tomatoes. Agricultural Research Service (ARS) in Illinois have genetically modified the fungus Fusarium sporotrichoides to manufacture lycopene using corn fibre material which is a leftover of the production of ethanol.

Timothy Leathers, a geneticist working at the National Center for Agricultural Utilisation Researche changed the metabolic pathway of natural trichothecene toxins of Fusarium sporotrichoides resulting in the production of lycopene in February 2004.

Tomato genome becomes fully sequenced with implication on related fruits and vegetable [169]

The full genome sequence of the tomato, Solanum lycopersicum, has been decoded and is published in May 2012 in the Journal Nature. Researchers aim to improve yield, nutrition, disease resistance, taste and colour of the tomato and other crops. The "Heinz 1706" tomato variety was used for the sequencing performed by the Tomato Genomics Consortium, an international collaboration between 14 countries.

Tomatoes possess some 35,000 genes arranged on 12 chromosomes. Sequencing of other tomato varieties can build on the data of the "Heinz 1706" tomato sequence reducing the costs. The Helmholtz-Zentrum provides the access to the tomato genome database at http://mips.helmholtz-muenchen.de/plant/tomato/index.jsp [170] James Giovannoni, leading the U.S. tomato sequencing team, says that the knowledge developed by the Tomato Genomics Consortium will help to look inside fruit biology, disease resistance, root development and nutritional qualities. Other plants, such as strawberries, apples, melons, bananas and many other fleshy fruits, share some characteristics with tomatoes. The new knowledge may be applied to these other plants. [171]

Genetically enhanced hybrid peppers

The robust pepper varieties, developed at the Hebrew University of Jerusalem, widen the ecological conditions under which the vegetable can be grown.

The new hybrid peppers, which come in various colours, have been raised to produce high yields under night time conditions as low as 10 degrees celcius. This is much lower than previous hybrids that required temperatures higher than 18 degrees celcius and needed costly heating to grow and develop.

The breeding project involved large-scale experiments with more than 25,000 plants a year, grown in target areas, mainly in the Arava region of southern Israel and the south of Spain. Dr. Elkind noted that vegetable production under mild winter conditions and using simple plastic or net protection is one of the most rapidly expanding, protective cultivation systems worldwide.

The major areas which use this production method, in addition to Israel and Spain, are Mexico and China.
Source: The Hebrew University of Jerusalem.

GM corn in France [172]

The study said that French Bt corn acreage is expected to boom from 500 ha in 2005 to 5,000 ha in 2006, as a result of the economic advantages experienced by Bt corn growers in 2005.

According to the Global Agriculture Information Network (GAIN), backed on French press reports, biotech corn cultivation is increasing rapidly, since 2005. Marie-Cécile Hénard writes is this report that pervasive presence of the European corn borer in Southern France provides strong incentive for further expansion. Bt corn provides an effective and profitable remedy against the European corn borer in this region.

French consumer resistance leaded by José Bové and other opponents of GM crops in France tried to to destroy GM fields. José Bové characterized commercial GM corn grown in France as "outlaw crops".

The findings of this pro-GM study suggest that the historical rejection of GM technology in Europa is on the wane. The WTO of course famously ruled earlier this year that Europe had violated its trade rules by banning GM food imports between 1999 and 2003.

The EU's approach to GM food regulation was criticised by pro-GM lobby for having little theoretical basis, and pandering to the fears and prejudices of its citizens. The EU's precautionary principle says that regulators should err on the side of caution, assuming that a prevention strategy is always appropriate.

Drought and fungal resistant GM wheat

The BASF plant biotechnology company, together with Australian research centre Molecular Plant Breeding CRC, CropDesign, a Belgian biotech company, that specialises on traits for yield-enhancement, drought tolerance and improved nutrient use efficiency of crops such as corn and rice.

These companies have built in 2006 the platform for the development of crops with higher yields, drought tolerance and resistance to fungal diseases.

Corn and wheat are the most important cereals. Drought and fungal diseases reduces yields in arid areas, Australia and Europe. These new breeds will help to overcome these serious crop losses.

Romania and GM soybean

Romania is growing large quantities of GM soybeans. A large part of which is exported to the European Union for use in animal feed. According to Simon Barber Romanian farmers would be in competitive disadvantage versus Brazilian, Canadian and US soybean suppliers, who are already exporting this same product to the EU. [173]

GM Maize

Monsantos Maize MON 810 had been approved before the EU biotech ban in 1998. Mon 810 has been assessed as safe for human health by the EU Commission. In conclusion, the Panel considers that the information available for MON 863 x MON 810 maize addresses the outstanding questions raised by the Member States and considers that it will not have adverse effects on human and animal health or the environment in the context of its proposed use.

In 2004 17 different seed strains of this maize have been allowed to be sold and grown in the EU.
Greece, however, held its ban despite not having provided any validated scientific evidence to support a ban, appealing against the EC's order at the European Court of Justice, the highest EU court. [174]

Luxembourg, Greece and Austria consistently vote against GMO approvals, while the UK, Finland and the Netherlands almost always vote in favour of any new GM authorisation.

Friends of the Earth (FoE) supports the biotech ban of Greece claiming that Monsanto's Roundup Ready soybeans, the most extensively grown GM crop today, has led to an increase in herbicide use. Independent reports from the US show that since 1996, GM corn, soybean and cotton have led to an increase in pesticide use of 55 million kilos.[175]
Juan Lopez of Friends of the Earth International said in Kuala Lumpur that the safety of GM crops cannot be ensured, that they are neither cheaper nor higher quality and that they are not the magical solution to solve world hunger. [176] [177]

The International Service for the Acquisition of Agri-biotech Applications (ISAAA) argue that the benefits of the technology to the food industry have simply become irresistible. The ISAAA is a not-for-profit organization that delivers the benefits of new agricultural biotechnologies to the poor in developing countries aiming to share these technologies to those who stand to benefit from them and at the same time establish an enabling environment for their safe use.

Emerging economies such as China, Brazil, Argentina and India are likely to be the greatest growth regions for the use of biotech foods Iran started to plant biotech rice in 2005 as being the first country to do so. [178]

According to ISAAA for the first time the absolute growth in the biotech crop area between 2003 and 2004 was higher in the developing countries (7.2 million hectares) than in industrial countries (6.1 million hectares). The organization highlites that the percentage growth was almost three times as high (35%) in the developing countries of the South, compared to the industrial countries of the North (13%). [179]

Testing for unapproved Genetically Modified Organisms rice BT63 will enter into force on 15th May [180]

The European Commission decided today to require compulsory certification for the imports of Chinese rice products that could contain the unauthorised GMO Bt63. The decision has been taken after rice products -originating in, or consigned from, China and containing the unauthorised genetically modified rice "Bt 63" were discovered in the EU market between 2006 and 2007.

The presence of the unauthorised Bt63 was first discovered in the United Kingdom, France and Germany in September 2006. Despite measures announced by the Chinese authorities in 2007, alerts concerning the presence of the unauthorised genetically modified rice "Bt 63" were reported until late 2007.

EU emergency measures

The emergency measures adopted by the Commission mean that, as of April 15, only consignments of the rice products indicated in a specific Annex of the Decision can enter the EU. These consignments must be tested by an official or accredited laboratory using a specific testing method and accompanied by the analytical report assuring they do not contain Bt63.

Responsible parties for the controls

China is responsible for ensuring that Bt63 does not enter the EU food chain and that imports are certified as free from this unauthorised GMO. Member State authorities are responsible for controlling the imports at their borders and for preventing any contaminated consignments from being placed on the market. In addition, they should carry out controls on products already on the market, to ensure that they are free from Bt63.

Business operators importing rice products from China are also responsible for ensuring that Bt63 does not enter the EU food chain and that imports are certified as free from this unauthorised GMO, in accordance with the EU food law principle that operators are responsible for the safety of the food or feed that they place on the market.

Non-authorised genetically modified rice "Bt63" food product in Finnland detected [181]

EVIRA, the Finnish Food Safety Authority reports that samples of rice vermicelli were found to contain the non-authorised genetically modified rice "Bt63". The product was withdrawn from the market and destroyed. One soy bean jelly sample contained genetically modified soy in excess of the 0.9% labelling limit. The importer was ordered to change the labelling of the product. The expression "genetically modified" or, e.g., "prepared from genetically modified soy" must appear in the declaration of ingredients right after the genetically modified ingredient if it exceeds 0.9%. [182]

Food products containing soy and maize (e.g. tofu products and precooked food products) as well as from Chinese and American rice products, and organic foods were included in sampling plan. All the analysed organic samples tested negative for GM ingredients.

Bayer CropScience puts rice industry in distrust [183]

According to Green Peace a series of scandals erupted in 2006 as world rice supplies were discovered to be contaminated with unapproved genetically engineered (GE) rice varieties. Field trials of GE rice in the US and the illegal sale of rice seed in China led to unapproved GE rice entering global food supply chains. Contaminated food stocks were found and pulled from shelves in European stores. Widespread bans on US-produced rice were enacted.

The latest GE contamination scandal shows that once GE organisms are released into the environment, the consequences for consumers, farmers and traders are enormous. As a result, farmers, millers, traders and retailers around the globe are facing massive financial costs, including testing and recall costs, cancelled orders, import bans, brand damage and consumer distrust-distrust that could last for years.

Lawsuits have been filed by US rice farmers against Bayer CropScience already, as farmers struggle to protect their livelihoods from GE contamination. On the other side,WTO has published a ruling on a case brought against the EU by the US, Canada and Argentina over Europe imposing restrictions on the importing of GE food.

BASF and Monsanto are united in plant biotechnology [184]

The two giants wich dominate agricultural products joined at March 21, 2007 The cooperation will focus on the development of crops with higher yields and crops that lead to consistent yields under adverse environmental conditions, such as drought.

The most promising candidate genes of both companies will be advanced for accelerated joint development and for commercialization by Monsanto.

The two companies expect to generate a greater number of viable research projects than they could have done on their own, accelerate the development of new products, and bring a greater number of traits to the market at a faster speed.
Monsanto will receive 60 percent of net profits and BASF will receive 40 percent of net profits.

US court prohibits GMO sugar beet from Monsanto [185]

United states sugar beet are 95% genetic modified plants. Genuity Roundup Ready sugarbeets were introduced to farmers during the 2008-2009 crop season. A court decided now that the US government authorised the planting of the GMO sugar beet of Monsanto before environmental impact statement had been completed and finally approved.

More than half of sugar produced in USA comes from sugar beet. The court denied further use of the GMO seed. Growers must use former not GMO seeds, starting in 2011.

Monsanto fears a drop of two billion Dollar of revenues in 2011 and 2012 due to this prohibition. It may take years to get a new approval for the GMO sugar beet. BASF works together with Monsanto developing GMO seeds of corn, rape, wheat, soy and cotton. BASF developed GMO sugar beet plants with KWS company.

Stem cell lines from bovine embryos

Dr. Paul Verma from the Dairy Cooperative Research Centre (CRC) made researches on the multiplication of bovine ES-like cells stem cell lines from bovine embryos.

Results from this research have many implications for the application of advanced breeding technologies in dairy cattle and other livestock. Advanced stem cell technologies offer the opportunity to improve performance of IVF-based animal breeding systems (in vitro fertilisation). In the dairy industry, this can help to shorten the interval between generations of animals and speed up the entire process of genetic improvement.

This system requires the use of embryos rather than semen in artificial breeding technology. Molecular diagnostics are being developed to allow precise monitoring of genes in embryos before they are implanted.

According to CRC the monitoring technology allows the healthiest, most viable, embryos to be identified and implanted which will improve success rates and lower costs. [186]

Kanamycin resistant potato has no adverse effect on human health

The Scientific Panel on Genetically Modified Organisms (GMO Panel) of the European Food Safety Authority (EFSA) issued in 2005 an opinion on genetically modified potato EH92-527-1 (Unique identifier BPS-25271-9), with an altered starch composition (higher amylopectin:amylose ratio).

The BASF Plant Science GM potato EH92-527-1 is derived from the cultivar Prevalent. Potato leaf discs were transformed by Agrobacterium-mediated gene transfer technology using kanamycin resistance gene (nptII) as selectable marker. The modification involves inhibition of the expression of granule bound starch synthase protein (GBSS) responsible for amylose biosynthesis. The starch produced has little or no amylose and consists mainly of amylopectin. Amylopectin starch potatoes are mainly used for the production of starch for industrial purposes, and the potatoes are not intended for direct human consumption. However it might also be used as food.

Potato rarely survives outside the cultivated environment and there is no indication of enhanced weediness or invasiveness of potato EH92-527-1. Potato has no cross-compatible wild relatives in Europe. Since the natural exchange of genetic material is only possible with other varieties of potato, there is negligible risk to the environment of any transgene flow. Therefore, no unintended environmental effects due to the establishment and spread are anticipated.

In the unlikely event that horizontal transfer of gene sequences would occur between the GM potato and bacteria, the bacteria would not pose any additional risk to human health or the environment.

No adverse effects on plant-associated organisms and soil function have been observed or would be likely from cultivation of the potato EH92-527-1. In addition, the GMO Panel agrees with the approach proposed by the applicant in the environmental monitoring plan.

EFSA found that the BASF GM potato is unlikely to have an adverse effect on human health or the environment in the context of its proposed uses. [187]

BASF GM potatoes release between 2007 and 2011 in Germany [188]

The Federal Office of Consumer Protection and Food Safety (BVL) approved the release of genetic modified potato, developed by BASF Plant Science in ten cultivation areas in Germany between 2007 and 2011. The approval is extended all over Germany, including Bavaria. The BVL does not expect any safety incidences which might endanger man and animals.

The marker gene used in these potato varieties, makes them resistant against the antibiotic kanamycin. The European Food Safety Authority has already identified no safety concerns which may have an adverse effect upon approval.

Measures to avoid spread out of the GM potato imposed by BVL

- The safety distance to conventional potato crops should be 10 metres.
- The aerial part of the plant must be destroyed chemically or physically before the harvest of the GM potato.
- The crops must be identified.
- The cultivation areas must be monitored for new breeds one year after GM use, if GM potatoes are found, the monitoring must be extended for another year.

The gene of the tale cress weed Arabidopsis thaliana together with a gene of a wild potato changes the starch composition of the potato . 677 strains of GM potatoes will be tested. Some strains will express amylose, other will express only amylopectine. They are tolerant to the herbicide Imazamox.

BASF GE potato field trial considered as illegal in The Netherlands [189]

On 7 March 2007, The Council of State in The Netherlands judged that the field trials of BASF concerning three GM potato herbicide-resistant varieties, two with changed starch content and one with high late blight resistance, were illegal and destroyed the permits immediately.


Intragenic Ranger Russet potatoes

[190]
Compared to more commonly used Russet Burbank, which is already 131 years old, the Ranger Russet has superior yields and greater disease resistance. This modified variety may also be interesting for Europe due to its excellent adaptability.

Caius Rommens indexRommens, Caius and coleagues reduced the expression simultaneously of the Ranger Russet's tuber-expressed polyphenol oxidase (Ppo), starch-associated R1, and phosphorylase-L (PhL) genes. For these changes no foreign DNA was introduced. The researchers say that French fries made from these potatoes contained about one-third the acrylamide content of control fries.

According to the authors the Ranger Russet presented increased black spot bruise resistance, reduced cold-induced sweetening, reduced stress-induced sugar ends, enhanced fry aroma, reduced amounts of processing-induced acrylamide, reduced starch phosphate content, and increased starch.

Genetic modulation of aroma of fruits and vegetables [191]

Dong-Sun Lee and colleagues 2008 used genetic manipulation of enzymes in plants to change aromas in fruit and vegetable. This may enable scientists to change the flavour and taste of fruits and vegetables. The authors say that the flavour of GM olive oil may be altered modifying the activity of enzymes which generate the substances responsible for flavour.

The research based on changes of the allene oxide synthase (AOS) into hydroperoxide lyase HPL. This gene modifies the enzymes which produce green leaf volatiles (GLVs), working with Arabidopsis thaliana, and used 3-D images of the enzymes to make a small, but specific, genetic change in AOS, leading to the generation of HPL.

The AOS enzyme produces jasmonate, the scent of jasmine flowers, and the hydroperoxide lyase HPL aromas to fruits and vegetables, GLVs.

The authors stress that this knowledge may be used for pest control. GM modification of enzyme pass could create pest resistant plants because green leaf volatiles and jasmonates also ward off predators, attracting predators of plant predators.

Unwanted gene flow from canola

[192] Unwanted gene flow from herbicide resistance canola can occur between canola varieties over large distances, but as isolated events.

Janine Baker and Christopher Preston came to the conclusion that insect mediated pollen transfer accounts for the low levels of gene flow that have been observed to occur sporadically at long distances from the source.

The scientists sprayed bees with fluorescent paint and determined the level of their movement between canola fields.They found that honeybees tended to return to the same field when resources were abundant, but that they foraged over larger distances when resources were scare or patchy. These findings suggest that the honeybee has the potential to contribute to gene flow over large distances but at a very low frequency.

European court ruled that GM contaminated honey must be labeled and needs EU food safety approval [193]

German beekeepers sued the Bavarian government after their honey was contaminated by field trials of Monsanto's GM maize Monsanto 810 in 2005. The GM maize is used as feed and is not allowed for the consumption by humans. Any GM contaminated honey must now be subject to full safety authorisation and labelling.
Half of Canadian and South American honey imports must be removed from self of European stores: Half of imported honey from Canada and South America have been tested positive for GM contamination by Ökotest in 2009. Contaminated products must be taken out of the shelf of European food stores. This will have an effect on European GM ruling and is a severe setback for the GM agrarian lobby which is trying to water down the zero tollerance to GM contamination of food. Honey producers can now sue any maize GM grower when his honey is contaminated by Monsanto 810 polen or othes GM variety.

In 1998 Monsanto obtained authorisation to market certain MON810 maize products as maize flour, maize starch, and maize oil but not for MON810 food products in general. Honey is a special product because besides the different kinds of sugar produced by the bees it also contains pollen. Therefore it is in parallel an animal and plant product. The GM regulation 1829/2003 excludes animal products from obligatory GM labelling, if animals are fed with GM feed. [194]

Increasing likehood of proliferation of GM food

The WTO has ruled in favor of the US challenge of the European Moratorium on approvals of agricultural crops preliminary decision by a World Trade Organization (WTO) panel on the U.S. challenge of the European Union (EU) moratorium on approvals for crops derived from biotechnology.

The United States, joined by Argentina and Canada in the challenge, had asserted the moratorium violates international trade rules and undermines the development and use of biotechnology. They had argued the ban was not based on scientific evidence and maintained that biotech crops are as safe to health and the environment as other crops. All parties now will have a chance to review and comment on the preliminary ruling, which was released February 7 to the U.S. and EU governments and subsequently leaked to the press.

The WTO likely will issue a final decision on the approval challenge in late 2006 or early 2007, a U.S. trade official said in advance of the decision. The loss of U.S. agricultural sales to Europe because of the ban has amounted to "several hundred millions of dollars" annually, the official said. The U.N. Food and Agriculture Organization (FAO) and World Health Organization (WHO) also have said there are no greater risks associated with biotech-derived foods than with conventional plants and foods.

Resistance to genetically altered crops among consumers in Europe remains strong, therefore the lifting of the ban might not have a significant effect on the quantity of U.S. agricultural exports to the region. [195]

WTO Dispute Settlements, Reports out on Biotech Disputes [196]

The WTO, on 29 September 2006, issued the reports of the panel that had examined complaints by the United States, Canada and Argentina, respectively, against "European Communities - Measures affecting the approval and marketing of biotech products" (DS291, DS292 and DS293).

In this report the panel concluded that the European Communities has acted inconsistently with its obligations under the SPS Agreement in respect of the relevant member State safeguard measures, it must be presumed to have nullified or impaired benefits accruing to Argentina under that Agreement.

In the light of these conclusions, the Panel recommends that the Dispute Settlement Body request the European Communities to bring the relevant member State safeguard measures into conformity with its obligations under the SPS Agreement.

No repercussion on the European GMO-regulation is expected. [197]

Improving citrus breeding

The University of California, Riverside is one of the few universities in the world to support an active citrus breeding program. Professor of Genetics Mikeal Roose developed the GeneChip Citrus Genome Arrayin classifying the citrus genes that are associated with taste, acidic content and disease. The citrus array will be used to develop new diagnostic tools for the improvement of citrus agriculture and post-harvest fruit handling, as well as to understand mechanisms underlying citrus diseases. [198]

Citrus fruits are abundant in limonoids which are a class of chemically related compounds present in citrus fruit. Limonoids have been shown to have anti-cancer activity and can inhibit breast cancer cellular proliferation in estrogen receptor negative (ER-) and positive (ER+) cells.

The researchers found that limonoids, particularly limonin, slowed tumor growth and were able to prevent or delay the regrowth of resected tumors in these models. Limonoids also reduced the incidence of lung metastases. These results suggest that citrus juice components may be beneficial in the prevention of mammary cancer. [199]

Other benefits of citrus juice [200]

Consumption of 750 mL but not of 250 or 500 mL orange juice daily increased HDL-cholesterol concentrations by 21% , triacylglycerol concentrations by 30%, and folate concentrations by 18%; decreased the LDL-HDL cholesterol ratio by 16%; and did not affect homocyst(e)ine concentrations. Plasma vitamin C concentrations increased significantly.

Kurowska and colleagues found in 2000 that orange juice (750 mL/d) improved blood lipid profiles in hypercholesterolemic subjects, confirming recommendations of the US food guide pyramid to consume about 5-10 servings of fruit and vegetables daily.

EuropaBio

EuropaBio, the European Association for Bioindustries, has 50 direct members operating worldwide and 25 national biotechnology associations representing some 1500 small and medium sized enterprises involved in research and development, testing, manufacturing and distribution of biotechnology products. http://www.europabio.org

Agricultural Biotechnology International Conference (ABIC)

The Foundation was set up as a not-for-profit corporation, based in Saskatoon, Saskatchewan. The Foundation's goal is to ensure ongoing opportunities for continuous learning and networking within the agbiotech community through the Agricultural Biotechnology International Conference.

The Foundation performs advisory, promotional and supportive functions for the conferences. As the Foundation shapes the ABIC concept, it strives to position the Conference at the forefront of agbiotech innovation.

The Conference on Co-existence between GM and Non GM crops in Vienna 2006 forms part of the Commission's consultations with interested parties on the development of efficient and cost-effective strategies to ensure co-existence.

Agriculture is an open process, which means that perfect segregation of the different agricultural production types is not possible in practice. Co-existence of these production types which will not lead to a systematic exclusion of one or more of them can only be ensured if the segregation measures are designed in a way that takes these limitations into account.

Co-existence thus concerns only the economic implications of GMO admixture, the measures to achieve sufficient segregation between GM and non-GM production and the costs of such measures. [201] [202] [203]

It is already completely legal to grow certain GM crops within the European Union, with this list of permitted crops likely to get longer.
On 23 July 2003, the European Commission adopted Recommendation 2003/556/EC on guidelines for coexistence, reaffirming that measures for coexistence should be developed by the Member States. The guidelines specify that those farmers who introduce the new production type in a region should bear responsibility for implementing the farm management measures necessary to limit gene flow. Following these guidelines, measures currently being discussed by Member States are designed to be taken by GM crop farmers. Furthermore, since seeds may be a source of adventitious GM presence in agriculture, the European Commission initiated discussions on setting

specific thresholds for the adventitious presence of GM seeds in conventional seeds, lower than those allowed in the final crops (0.9%). Therefore, seed production might have to operate under different coexistence requirements than crop production. These discussions are still ongoing. [204]

In 1998 Spain approved the first commercial growing of the transgenic maize BT 176 from Ciba Geigy (now Syngenta). The Spanish government continues to allow transgenic maize growing although there is scientific uncertainty over its safety.

NGOs fear widespread contamination

Consumer organizations fear GM crops will cause widespread contamination by genetic modified with heavy consequences for organic growers such as Enric Navarro, an organic maize corn grower from Girona in Spain claiming that in 2005 his crop was contaminated up to 12.6 per cent by GM maize. He burnt the crop at a massive economic loss."This year I will again sow maize. But if it is again contaminated (with genetically engineered maize) I will abandon the growing of organic maize. It is obvious that I cannot carry on my shoulders the episodes of contamination like these, year after year." [205]

Jose Bové [206]

The anti-globalisation leader Jose Bové combats junk food, US trade tariffs and the risks to the environment of genetically modified organisms. He was arrested and sentenced several times. He was prevented to enter Mexico to attend a WTO summit in Mexico in 2003. He was deported from Hong Kong on WTO talks and prevented to enter the US to speak at a conference at Cornell University in New York state in 2006. On april 2006 Jose Bové, was arrested attempting to occupy Monsanto's seed facility near Carcassonne in southern France. The action had been organised by the Collectif des faucheurs volontaires, la Confederation paysanne and Greenpeace.

According to Olivier Keller, national secretary of the Confederation Paysanne, GM is harming the environment and is causing genetic contamination of the food-chain and agriculture, thus threatening the right of farmers and consumers to grow and eat GM free food.

Biotech companies and regulators argue that adequate controls are already in place to ensure that the cross-contamination is not an issue.

Simon Barber, director of the plant biotechnology unit at EuropaBio - the European association for bioindustries counters that thousands of European farmers grew GM last year, successfully co-existing with their neighbours.

According to anti-GM campaigners GM crops will cause widespread contamination, leaving consumers with no GM-free choice at all. Pro-GM forces on the other hand argue that consumers must be given the choice, and that the WTO ruling backs this up. But opposition to GM food will scarcely be long lasting.

Europe's food safety authority announced that the GMO Panel is of the opinion hat there is no reason to believe that the continued placing on the market of Bt176, T25 and MON810 maize, and Ms1xRf1 and Topas 19/2 oilseed rape is likely to cause any adverse effects for human and animal health or the environment under the conditions of their respective consents," said an EFSA statement this week.

Future of biotechnology and BASF

Dr Hans Kast, president and CEO of BASF Plant Science announced in 2006 that BASF has identified plant biotechnology as the largest of five key future-growth clusters. BASF has built its work on the cooperation of Metanomics and SunGene companies.

Metanomics covers BASFs gene-discovery research identifying the metabolic functions of each and every plant gene.Together with SunGene company the gene-mapping knowledge is being applied to optimise and control specific traits in a plant Increasing the content of valuable compounds in plants, such as vitamins, carotenoids and proteins, such as NutriDense, the nutritionally enhanced corn for the feed industry. It has higher content of protein, essential amino acids, oil/energy and more available phosphorous.

Simon Barber, European Association for Bioindustries, EuropaBio argues that co-existence between GM and non-GM crops has been a success in Europe for years and there is no substantiated evidence to the contrary. According to Barber In Europe alone, there are thousands of farmers and dozens of scientists with real field experience of successful co-existence who were not asked to share their knowledge at this conference. [207]

EU concept of coexistence

[208] The cultivation of genetically modified organisms (GMOs) in the EU is likely to have implications for the organisation of agricultural production. On the one hand, the possibility of the adventitious (unintended) presence of genetically modified (GM) crops in non-GM crops, and vice versa, raises the question as to how producer choice for the different production types can be ensured. In principle, farmers should be able to cultivate the types of agricultural crops they choose, be it GM crops, conventional or organic crops. None of these forms of agriculture should be excluded in the EU.

On the other hand, the issue is also linked to consumer choice. To provide European consumers with a real choice between GM food and non-GM food, there should not only be a traceability and labelling system that functions properly, but also an agricultural sector that can provide the different types of goods. The ability of the food industry to deliver a high degree of consumer choice goes hand in hand with the ability of the agricultural sector to maintain different production systems.

Coexistence refers to the ability of farmers to make a practical choice between conventional, organic and GM-crop production, in compliance with the legal obligations for labelling and/or purity standards.

The adventitious presence of GMOs above the tolerance threshold set out in Community legislation triggers the need for a crop that was intended to be a non-GMO crop, to be labelled as containing GMOs. This could cause a loss of income, due to a lower market price of the crop or difficulties in selling it. Moreover, additional costs might incur to farmers if they have to adopt monitoring systems and measures to minimise the admixture of GM and non-GM crops. Coexistence is, therefore, concerned with the potential economic impact of the admixture of GM and non-GM crops, the identification of workable management measures to minimise admixture and the cost of these measures.

Economic aspects of coexistence versus environmental and health aspects [208]

It is important to make a clear distinction between the economic aspects of coexistence and the environmental and health aspects dealt with under Directive 2001/18/EC on the deliberate release of GMOs into the environment.

According to the procedure laid down in Directive 2001/18/EC, the authorisation to release GMOs into the environment is subject to a comprehensive health and environmental risk assessment. The outcome of the risk assessment can be one of the following:
- a risk of an adverse effect to the environment or health that cannot be managed is identified, in which case authorisation is refused,
- no risk of adverse effects on the environment or health is identified, in which case authorisation is granted without requiring any additional management measures other than those specifically prescribed in the legislation,
- risks are identified, but they can be managed with appropriate measures (e.g. physical separation and/or monitoring); in this case the authorisation will carry the obligation to implement environmental risk management measures.

If a risk to the environment or health is identified after the authorisation has been granted, a procedure for the withdrawal of the authorisation or for modifying the conditions of consent can be initiated under the safeguard clause set out in Article 23 of the Directive.

Since only authorised GMOs can be cultivated in the EU(1), and the environmental and health aspects are already covered by Directive 2001/18/EC, the pending issues still to be addressed in the context of coexistence concern the economic aspects associated with the admixture of GM and non-GM crops.

1.3. The Round Table on coexistence

A Round Table to examine the latest research results on the coexistence of GM and non-GM crops was hosted by the European Commission in Brussels on 24 April 2003. It focused on coexistence issues raised by the introduction of GM maize and GM oilseed rape into EU agriculture. Expert panels presented the scientific findings, which were then discussed with a range of stakeholders representing the farming sector, industry, NGOs, consumers and other players.

The Round Table sought to provide a scientific and technical basis, drawing on practical farming experience, for whatever agronomic and other measures may become necessary to facilitate the sustainable coexistence of these different agricultural production types.

The present guidelines draw on the results of the Round Table, a summary of which, prepared by a group of participating scientists, is available on the following Internet site
http://www.gmo-compass.org/pdf/law/guidelines_coexistence.pdf

EFSA guidance on environmental impact of GM plants [209]

The European Food Safety Authority (EFSA) has published updated guidance for the environmental risk assessment (ERA) of Genetically Modified (GM) plants focused in particular on data generation, collection and analysis and the evaluation of possible long-term effects of GM plants and potential effects on non-target organisms, such as those insects which are not meant to be the target of the toxin produced by some GM plants.

The Guidance covers persistence and invasiveness of the GM plant, possible gene transfer from the plant to micro-organisms, how the GM plant could affect cultivation, management and harvesting techniques, and effects on human and animal health.

In order to assess the safety of a GM plant EFSA requires all applicants to follow its guidance documents which specify the type of data and information that should be submitted. Seven specific areas were reviewed which include the persistence and invasiveness of the GM plant, considering the possible plant-to-plant gene transfer; the likelihood and consequences of gene transfer from the plant to micro-organisms; the potential evolution of resistance in target organisms; the potential effects on non-target organisms; the biogeochemical processes, such as changes in soil composition, and the potential impact of the cultivation, management and harvesting techniques of the GM plant.

The guidance document includes detailed requirements for: the choice of appropriate non-GM comparators with which the GM plant is compared during the safety evaluation. Also types of receiving environments have to be considered; long-term effects and the experimental design of laboratory and field studies; and their statistical analysis are defined in guidance.

Amendment of Directive 2001/18/EC liberalising GMO cultivation in Europe [210]

In July 2010, the Commission adopted a proposal which amends Directive 2001/18/EC, giving members states the freedom to allow, restrict or ban cultivation of GMO on their territory. The proposal, has to go through a co-decision process, however the plan is being disapproved by anti-GMO states and also pro-GMO states alleging fragmentation within the single market, and being incompatible with rules set by the WTO.


[211]
Crop antibiotic resistance Antibiotic resistance
  Marker used for selection Genes under bacterial
    promoters
Corn -- 3 nptII
    5 bla
    1 cat
Tomato 4 nptII --
Oilseed rape 3 nptII --
cotton 3 nptII 2 aad
Potato 2 nptII 1 aad
Soybean 1 nptII 1 bla
Squash 2 nptII --
Papaya 1 nptII tetR
Radicchio 1 nptII --

US FDA/CFSAN 1997

Green Fluorescent Protein [212]

Green Fluorescent Protein was discovered and developed by Martin Chalfie, Osamu Shimomura and Roger Y. Tsien as a companion protein to aequorin, the chemiluminescent protein from the jellyfish Aequorea victoria. It has become a valious marker of gene expression and protein targeting in intact cells and organisms, and may be used as physiological indicators, biosensors, and photochemical memories. The three researchers were awarded the 2008 Nobel Prize in Chemistry on 8 October 2008

It has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm. Its emission peak is at 509 nm which is in the lower green portion of the visible spectrum. GFP has a typical beta barrel structure, consisting of one beta-sheet with alpha helix(s) containing the chromophore running through the center. Inward facing sidechains of the barrel induce specific cyclization reactions in the tripeptide Ser65-Tyr66-Gly67 that lead to chromophore formation. This process of post-translational modification is referred to as maturation. The hydrogen bonding network and electron stacking interactions with these sidechains influence the color of wtGFP and its numerous derivatives. The tightly packed nature of the barrel exclude solvent molecules, protecting the chromophore fluorescence from quenching by water. [213]


Nonradioactive fluorescent biomarker substances for biomedical and research [214]

Fluorescent organic dyes are widely used as nonradioactive labels in biological analysis or as biomarkers in biomedical applications. Synthetic fluorescent molecules should exhibiting high extinction coefficients, high quantum yields, narrow emission bands, and photostability.

According to Song Lin Niu and colleagues 2009 available fluorescent substances have poor water solubility, and low resistance to the formation of nonfluorescent dimer and higher aggregates. The authors report two methods to introduce (poly)sulfonated linkers onto 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY) which are water-soluble.

The spectral properties of the resulting water-soluble BODIPY derivatives were found to present good characteristics under physiological conditions making them attractive as biomarkers.

Real-time gene expression Monitoring [215]

Grover and colleagues 2008 Researchers from the marked fruit fly genes with Green Fluorescent Protein and detected the green glow of the active gene with a camera equipped with a special filter. A second camera system tracked the movement of each fly in three dimensions. According to the authors genes which are responsible for certain behaviours may thus be tracked. The researchers hope to get new insights on ageing in the fruit fly and biological processes.

The authors note that two genes, hsp70 and hsp22, which respond to oxidative stress, spiked in the hours before the death of a fly. This suggests a sharp increase in oxidative stress as the fly began dying of natural causes. The phase of hsp gene expression relative to fly activity levels was different for each fly, and remained the same throughout the life span.

Linking Green Fluorescent Protein to any gene, its expression change may be monitored automatically and can be correlated with the animal activity, behavior and mortality.

Green Fluorescent Protein unveils details of the physiology of bacteria [216]

Sothward and Surette 2002 describe the use of the green fluorescent protein (GFP) from the jellyfish Aequorea victoria which can be expressed as a fluorescent protein in any other organism and became the most common reporters in use today. GFP is used for characterizing spatial and temporal dynamics of gene expression, protein localization and protein-protein interactions in living cells and bacteria.

Green Pigs [217]

Scientists from National Taiwan University's Department of Animal Science and Technology created pigs which are green through and through by adding genetic material from jellyfish into a normal pig embryo. The eyes, teeth and trotters of the pigslook green, and their skin has a greenish tinge. In the dark, a blue light makes them glow. The researchers hope the green pigs will mate with ordinary female pigs to create a new generation.

Shape and size gene of tomatoes identified which may also be applied in other plants [218]

Esther van der Knaap studied US the SUN gene from the tiny round wild ancestor and compare it with the actual elongated tomato varieties

Furthermore, the gene encodes a member of the IG67 domain of plant proteins, called IQD12 - a family of proteins whose discovery is relatively new.

The authors found that one of the major genes controlling the elongation of tomato, encode a member of the IQ67 domain-containing family, resulting in an unusual gene duplication event mediated by the long terminal repeat retrotransposon RIDER. This resulted in the increased fruit size and variation in fruit shape of domesticated tomatoes. Introducing the gene in the wild small tomato, the plant produced big, elongated tomatoes. Small original tomatoes were produced after removal of the gene.

The scientists hope to control the shape of other very diverse crops, such as peppers, cucumbers and gourds when more researches are done on this matter.

The Rx gene


Novel tomato genes affecting the functioning of the Rx gene [219]

Five tomato mutants of a transgenic, Rx1-expressing "Micro-Tom" line, were found by Sturbois et al 2012 to be affected in the Rx-mediated resistance against Potato virus X (PVX). The mutant tomatoes failed to develop lethal systemic necrosis upon infection with the virulent PVX-KH2 isolate and accumulation of the virus took place in all mutnats. However, the Rx1 resistance against the PVX-CP4 avirulent isolate was still effective, The mutants are not affected in the Rx1 transgene or in the Hsp90, RanGap1 and RanGap2, Rar1 and Sgt1 genes.

The mutants have retained the ability to mount an Rx1-mediated hypersensitive response. The authors stress that the five Micro-Tom mutants may be used to identify new tomato genes associated with the Rx gene.

In 1999 Bendahmane, Kanyuka and Baulcombe reported that Rx-mediated extreme resistance against potato virus X in potato does not involve a necrotic hypersensitive response at the site of initial infection. The authors suggest that the Rx gene differs from the usual disease resistance of plants, where both reactions occur simultaneously. In this study Rx gene of Nicotiana spp and potato were found to be able to initiate a cell death response but extreme resistance is separate and epistatic to necrosis. Epistatic means that the effects of the Rx gene is modified by one or several other genes called hypostatic.The authors concluded that cell death and pathogen arrest are separate disease resistance responses in plants. [220]

The potato (Solanum tuberosum) nucleotide binding-leucine-rich repeat immune receptor Rx confers resistance to potato virus X (PVX) and requires Ran GTPase-activating protein 2 (RanGAP2) for effective immune signaling. Coexpression with RanGAP2 intensifies the Rx-mediated immune signaling, leading to a hypersensitive response and amplified resistance to potato virus PVX. The protein RanGAP2 also stabilizes the Rx gene, contributing to defense signaling. In mutants the coexpression of Rx with a WPP domain of RanGAP2 leads to hyperaccumulation of both the WPP domain and Rx suppressing resistance to PVX and the hypersensitive response. RanGAP2 acts as a cytoplasmic retention factor for Rx, by interfering in the defence regulating partitioning of Rx. [221]


Genetic of Micro-Tom tomato cultivar [222]

The tomato cultivar Micro-Tom is a crossing Florida Basket and Ohio 4013- 3 cultivars, initially produced as ornamental plant. However, due to its small small size, rapid growth, and easy transformation it is used as model system for molecular research on the regulation of berry fruit development.

Martí et al. 2006 describe the genetics of Micro-Tom and report mutations in the SELF-PRUNING (SP) genes which generate a determinate phenotype, and mutations in the DWARF (D) genes which leads to mis-splicing and production of at least two shorter mRNAs. The existence of at least two resistance loci to the plant pathogen Cladosporium fulvum were reported.

The Micro-0 variety that is fully susceptible to Cladosporium fulvum is a mutation of the Cf03Micro-Tom. It is used for studies on Cladosporium fulvum resistance genes.

Based on the Micro-Tom variety, Saito et al. 2011 produced a family of mutant tomato population by ethylmethane sulfonate (EMS) mutagenesis and gam-ray irradiation.The mutant phenotypes were classified into 1,819 phenotypic categories in 1,048 mutants..To manage these data, the authors developed the TOMATOMA, a relational system interfacing modules between mutant line names and phenotypic categories, with open access at http://tomatoma.nbrp.jp/index.jsp. [223]

Organic potato production techniques [224]

Healthy soils, crop rotations and the help of natural predators are used by organic farmers to control crop pests and diseases in a sustainable agriculture. Few crop diseases, however, are so resistant to these techniques that additional chemique help is required such as spraying copper up to 6kg/ha per year together with crop rotation to control potato blight. Organic farmer say that copper does not build up in the soil, due to the crop rotations. Copper sprayed on leaves does not accumulate in tubers like other pesticides.

Health concerns related to GM potatoes and tomatoes [225]

Feeding trials by two scientific teams found that GM potatoes cause lesions in the gut wall of rats and mice. In October 1999 Ewen and Pusztai wrote that diets containing genetically modified potatoes expressing the lectin Galanthus nivalis agglutinin (GNA) had variable effects on different parts of the rat gastrointestinal tract, such as the proliferation of the gastric mucosa.

The biotechnology industry reacted very aggressively and tried to mobilise the scientific community to undermine the credibility of the work. Similar effects have been found with GM tomatoes in two US feeding trials, which found that GM tomatoes cause lesions in the gut wall of rats.

Drought resistant tomato [226]

Hirshi and colleagues studied the AVP1 (vacuolar H+-pyrophosphatase (H+-PPase))gene from Arabidopsis thaliana which results in enhanced performance under soil water deficits. According to their work, the AVP1 gene plays an important role in root development through the facilitation of auxin fluxes.

The scientists introduced The AVP1 gene in tomatoes resulting in more robust root systems allowing transgenic tomato plants to take up greater amounts of water during the imposed water deficit stress, resulting in a more favorable plant water status and less injury. The authors are working on how this technology could be applied to other commercial crops.


Abscisic acid, a plant hormone, may improve drought-resistance of crops [227]

Cutler, et al.2011 report further findings in abscisic acid , a plant stress hormone, which regulates drought-tolerance of crops. Plants, under drought stress produce abscisic acid which signals stomata on leaves to close, thus reducing water loss, cessation of plant growth to reduce water consumption and other tress responses.

The Study of Cutler and colleagues found how to increase the stress response pathway by modifying the abscisic acid receptors so that they can be turned on. The authors tested more than 740 variants of the stress hormone receptor, to find the best hormone receptor to improve crops at field.

The researchers focused on Pyrabactin resistance (PYR) 1 and its relatives which belongs to a family of soluble abscisic acid (ABA) receptors that inhibit type 2C protein phosphatases (PP2C) when in their agonist-stabilized conformation. Mutations that stabilize this agonist-bound conformation may be used to activate signalling in vivo.

The study improved the knowledge of PYR1 and related ABA receptors and found that activation of a single receptor is sufficient to activate signalling in plants to increase resistance to stresses such as drought, flooding, increased salinity and freezing temperatures. This is to increase yields under stress situations, however it may not green the desert Cutler cautioned.

Abscisic acid (ABA), also known as abscisin II and dormin. ABA functions in many plant developmental processes, including bud dormancy. Abscisic acid owes its names to its role in the abscission of plant leaves in preparation for winter.

Cutler et al 2010 studying the ABA signalling network, identified ABA receptors and developed a new model for ABA based on the soluble PYR/PYL/RCAR receptors function directly regulating PP2C phosphatases, which in turn directly regulate SnRK2 kinases. The authors suggests that future research should focus on the model of SnRK2s and PP2Cs, hormone interaction and other factors involved in ABA signalling. [228]

Autoactivation of abscisic acid (ABA) signaling SnRK2 kinases [229]

Ley-Moy Ng et al. 2011 explain that central for Abscisic acid (ABA) signaling is the ABA-mediated autoactivation of three monomeric Snf1-related kinases (SnRK2.2, -2.3, and -2.6). In the absence of ABA, SnRK2s are kept in an inactive state by forming physical complexes with type 2C protein phosphatases (PP2Cs).

The authors report the crystal structures of SnRK2.3 and SnRK2.6, and describe the ABA-mediated release of PP2C inhibition to activation of SnRK2 kinases, taking place in a two-step mechanism of intramolecular kinase activation.

Vitamin B1 (thiamine) participation in plant adaptations to certain stress, mediated by abscisic acid [230]

Rapala-Kozik et al. 2012 presented a model of the expression of thiamine biosynthetic genes, of genes encoding thiamine diphosphate-dependent enzymes and the levels of thiamine compounds responding to oxidative, salinity and osmotic stress. Also a regulatory role of abscisic acid in the stress sensing phase is being suggested.

The expression of genes involved in the thiamine diphosphate biosynthesis pathway, including that of THI1, THIC, TH1 and TPK were found to be up-regulated in stress response, correlating with increases in thiamine and its diphosphate ester content.

Among other stress phytohormones active, such as the salicylic, jasmonic and abscisic acids, only only abscisic acid significantly influence the expression of thiamine expression genes. Abscisic acid is important in the regulation of THI1 and THIC gene expression during salt stress but this process is more more complex and needs further clarification.

Exotic library [231] [232]

Actual crops resulted on a selective breeding from a very small number of wild plants. The genetic variation of wild plants is lost. Dani Zamir from Jerusalem University believes that exhausted genetic diversity presents a limitation for breeders to improve tomatoes.

He suggests to go back into the wild and cross domesticated crops with their wild relatives in order to gain access to useful traits, such as pest- or drought-resistance by developing exotic libraries Zamir explains that an exotic library consists of a large number of lines, each of which carries a small, well-defined chromosomal segment from the wild plant in the genome of the domesticated crop, and the whole genome of the wild relative is represented in a complete exotic library.

To develop drought-resistant tomatoes for the Middle East, Zamir crossed wild Lycopersicon pennellii tomato plants from the deserts of Western Peru, with a M82 high-yield tomato line of Israeli. In this way he developed a library of 50 introgression lines and crossed them again with three other lines and obtained a drought-resistant tomato with 30% higher yield than the original M82 line and does not rely on irrigation.

The delayed-ripening tomato FlavrSavr [233]

Artificial ripening with ethylene is used in 80% of US tomatoes. Conventional green tomatoes are harvested and transported. Before selling they are redened with ethylene. The delayed-ripening tomato Flavr Savr, a from Calgene was a genetically modified tomato that could be picked when ripe and transported without bruising. It carries the antibiotic resistance gene (Flavr Savr) for resistance to the antibiotic kanamycin. The enzyme polygalacturonase (PG), responsible for the tomato's softness, is supressed and the tomato keeps longer hard.

It was subject to heavy legal pressure from Monsanto and concerns about the use of antibiotic resistance gene. The tomatoes were only ever sold in a small number and then rapidly withdrawn.

GM tomatoes for puree

Zeneca developed at that time a GM tomato also based on the kanamycin antibiotic gene, for use in puree, a tomato to make it bulkier with reduced water content with increased viscosity.This tomato puree was sold starting in February 1996, attaining 60% of total sale of canned tomato puree. Due to safety concerns, both products were withdrawn from market.

There were concerns made public that the genes passed out of the tomatoes and entered bacteria, the bacteria could develop resistance to the antibiotic, undermining its medical effectiveness.

Other companies developing GM tomatoes

Agritope, Aventis, DNA Plant Technologies, Seminis and Monsanto.

FDA had decided that GM foods in general should not be regulated differently to non-GM foods and would not require pre-market approval. Unlike food additives, for which pre-market approval is required in the US, they argued that GM foods are the same or substantially equivalent to non-GM foods.

Safety concerns about Flavr Savr tomatoes

Calgene had fed rats with either a GM tomatoes. These studies revealed statistically significant differences between the effects of the GM and non-GM tomatoes. gross lesions, findings of gastric erosion or necrosis lesions and microscopic lesions were found in the rats were observed. The study methodology was argued FDA approved the GMO tomato.

Dr Joseph Cummins warned that the inclusion of a genetic sequence from the Cauliflower Mosaic Virus could create virulent new viruses and Dr Edwin Mathews wrote that the genetically modified plants could also contain unexpected high concentrations of plant toxicants.

The FDA was obliged to reveal these internal views in 1998 after a lawsuit filed by consumer groups.

The GM potatoes were rejected in the US in 2002 by US retailers and The British Retail Consortium has said UK supermarkets won't be stocking GM potatoes.

The high-flavolol tomato with chi gene from Petunia [234]

Shelagh R. Muir and colleagues modified tomatoes introducing a gene from Petunia encoding chalcone isomerase, an enzyme involved in flavonol biosynthesis has been used to produce transgenic fruit with an increase in peel flavonols of up to 78 fold, mainly due to accumulation of quercetin glycosides. The in vitro evidence for for One group of flavonoids, the flavonols (e.g., quercetin and kaempferol) are told to be cardiovascular protective.

Introducing the transgene gene (pBBC50) containing the Petunia hybrida chi gene under the control of the constitutive cauliflower mosaic virus (CaMV) double 35S promoter. into the skin of a tomato, Shelagh R. Muir and colleagues were able to increase flavonol production by up to 78 times. The taste was not affected, and 65% of the flavonols were retained when the GM tomatoes were processed into paste.
Shelagh R. Muir and colleagues concluded that constitutive overexpression of a Petunia gene encoding CHI in tomato resulted in elevated flavonol end products in the fruit peel. The tomato lines contained significantly increased levels of quercetin glycosides, and smaller but still substantial increases in kaempferol glycosides in fruit peel, offering potential health benefits.

USDA field test permission for genetically engineered crop

[235] The U.S. Department of Agriculture's (USDA) Animal and Plant Health inspection Service (APHIS) is responsible for protecting and promoting U.S. agricultural health, administering the Animal Welfare Act, and carrying out wildlife damage management activities.

The APHIS mission is an integral part of U.S. Department of Agriculture's (USDA) efforts to provide the Nation with safe and affordable food.

APHIS maintains the Biotechnology Regulatory Services (BRS) program regulating the field testing, movement, and importation of genetically engineered (GE) organisms that are known to be, or could be plant pests. BRS issues various types of permits for each of these activities. A Compliance & Enforcement program inspects, audits, and oversees activities under the permit process. BRS also evaluates petitions for deregulation to ensure that products being considered for removal from regulation do not pose a threat to U.S. agricultural or environmental health.

USDA APHIS protects America's agricultural and natural resources by ensuring the safe development of GE organisms using a science-based regulatory framework controlling :
- Permits for other types of genetically-engineered organisms include transgenic arthropods and applications for veterinary biologics.
- Permits for Non-genetically engineered articles include Plant pests, Plant and plant products, and Animal and animal products.

Biotechnology Regulatory Services (BRS)

Companies and organizations who wish to field test a genetically engineered crop, must obtain USDA permission through a permit. Companies must submit all plans for field testing for review by regulatory scientists who evaluate the risks of the test and the protocols to be employed. USDA will approve the plan if the proposed test conditions appear adequate to confine the regulated article within the field test site. To ensure compliance to the permitting conditions, field test sites are inspected and records are audited.

Depending on the nature of the genetically engineered crop, an applicant files either a notification or a permit application. In general, most of the plants are field tested under the notification procedure, a more streamlined approval process that is used only for familiar crops and traits considered to be low risk. Permitting is used for field tests of plants that have an elevated risk, such as plants producing pharmaceutical or industrial compounds.

APHIS also published a list of isolation distances from any contaminating source, valid for US, adapted from Table 5, 7 CFR part 201.76

Canada has its own list of isolation distances published under Canadian Food Inspection Agency (CFIA) APPENDIX 2: MINIMUM ISOLATION distances and periods of post-harvest land use restriction 2000. [236]

In Europe isolation distances between GM and non-GM fields are settled in the regulation 2003/556/EC.[208]

Discussion related to isolation distances [237]

According to Professor David James oil-seed rape separation distances should be applicable to any other crop. He ignores that rape is insect-pollinated and has fairly heavy pollen. Other plants like cereal crops are wind-pollinated traveling farther than rape pollen. (Val Spouge Braintree, Essex)

New tomatoes with increased soluble solids in the fruits [238]

According to professor of horticulture Avtar Handa of the Purdue University tomatoes with 10% increased soluble solids in the fruits have been geneticalliy engineered by its team. The fruits grow and ripen like traditional varieties.

Handa and colleagues constructed a backwards gene that inhibits the tomato ripening enzyme pectin methylesterase. This gene was introduced in an Agrobacterium which carried the gene to the DNA of the tomato. This backward gene resulted in higher soluble solids in the tomato making the production of thicker Ketchup possible.

According to Avtar Handa, crossing commercial varieties with wild species bearing the desired gene, takes years and may transfer unwanted characteritics. Genetic engineering creates the wanted tomato in less time compared with traditional cross- breeding.

Extended growing season for tomato [239]
Tomatoes are sensitive to frost. This shortens their growing season. Fish, on the other hand, survive in very cold water. Scientists identified a particular gene which enables a flounder to resist cold and used the technology of genetic engineering to insert this 'anti-freeze' gene into a tomato. This makes it possible to extend the growing season of the tomato.

Cross the species boundary [240]

While there may be risks associated with transferring undesirable traits through conventional breeding, a major concern about gene technology is not with the crossing of two of the same plant species, but the transfer of genes from one species, for example a fish, into another species such as a tomato, or a bacterium into a plant.

This ability to "cross the species boundary" through genetic engineering introduces an additional uncertainty and potential for serious harm.

One major area of concern was the gene crossover, sometimes described as transgenic, from one species to another. There was much less concern about wheat genes being used in wheat than bacterial genes being used in wheat for example. The use of viral promoter genes was a cause of even graver concern, in particular what might be the consequences of viral changes in subsequent generations. The Committee was told that little to no research had been done on later generation viral consequences.

Assurances that there is "no evidence" of harm may in fact mean no research has been done, and that worries the community. While there may be genetic exchange between species occurring in nature, genes from fish do not get into tomatoes under normal circumstances.

Opponents have argued that while the products of gene technology, such as herbicide resistant crops, long shelf life melons and delayed ripening tomatoes, are likely to bring some benefits to consumers, these products have been mainly developed to meet the needs of those in the food supply system, growers, transporters, wholesalers and retailers.

Notably, the crops that have been subject to genetic engineering are those that are economically important in the industrialised not the developing nations, for example maize, oilseed rape (canola), sugarbeet, tomato and potato. Nevertheless some research and trials have been conducted on wheat, rice, and cassava, an important food source in African and South American countries.20 Additionally, the main applications of genetic modification are producing herbicide and pesticide resistant plants, with much of the benefit going to the producers rather than consumers.

Recommendations from the Canadia Canola Council Canola to farmers

[241] The Canola Council of Canada is a non-profit association representing the entire canola industry - growers, input suppliers, researchers, crushers, processors, exporters and marketers.

The Canadian Canola-Council urges farmers to protect the reputation of canadian canola as a safe and high quality product stressing not to grow the canola varieties:

The genetic modified oilseed rape T45 from Bayer CropScience receives import green lights for food and feed uses, cultivation is excluded in EU [242]

The GMO Panel of the European Food Safety Authority is of the opinion that the molecular characterisation of the DNA insert and flanking regions of oilseed rape T45 does not raise any safety concern, and that sufficient evidence for the stability of the genetic modification was provided.

The GMO Panel is of the opinion that the composition of oilseed rape T45 does not deviate from that of conventional oilseed rape varieties, except for the introduced trait.

The PAT protein induced no adverse effects in acute dose oral toxicity studies in rodents. In addition, the PAT protein is rapidly degraded in simulated gastric fluid and inactivated during heat treatments.

A 42-day feeding study with broilers did not indicate differences in the nutritional value of T45 oilseed rape versus the non-GM comparator and confirms the nutritional equivalence of T45 oilseed rape containing diet in comparison with a conventional diet in broiler chickens.

The applications for oilseed rape T45 concern food and feed uses, import and processing of oilseed rape T45 and all derived products, but excluding cultivation of the crop in the EU. There is therefore no requirement for scientific assessment of possible environmental effects associated with the cultivation of oilseed rape T45. There are no indications of increased likelihood of establishment or survival of feral oilseed rape plants in case of accidental release into the environment of oilseed rape T45 seeds during transportation and processing.

However, the GMO Panel advises that appropriate management systems should be in place to minimise accidental loss and spillage of transgenic oilseed rape during transportation, storage, handling in the environment and processing into derived products.

In conclusion, the GMO Panel considers that the information available for oilseed rape T45 addresses the scientific comments raised by the Member States and that the GM oilseed rape T45 is as safe as its non genetically modified counterpart with respect to potential effects on human and animal health or the environment. Therefore the GMO Panel concludes that oilseed rape T45 is unlikely to have any adverse effect on human or animal health or on the environment in the context of its intended uses.

GM glufosinate-tolerant oilseed rape from Bayer Crop Science is safe [243]

The EFSA GMO Panel reiterates on the 17.09.2009 the previous conclusions that GM glufosinate-tolerant oilseed rape Ms8, Rf3 and Ms8 x Rf3 is unlikely to have an adverse effect on human and animal health or, in the context of its proposed uses, on the environment. This also applies to the products which are the subject of the present application.

Tomato mapping [244]

The genus Lycopersicon includes the cultivated tomato (L. esculentum Mill.) together with its wild relatives. The wild species bear a wealth of genetic variability. Less than 10% of the total genetic diversity in the Lycopersicon gene pool is found in L. esculentum (Miller and Tanksley, 1990). The center of diversity for tomato is located in western South America, and the cherry tomato L. esculentum var. cerasiforme is considered as the most likely ancestor of cultivated tomatoes. Karyotypes of the Lycopersicon species are very similar with little or no structural difference among species (Barton, 1950). As a crop plant, tomato is one of the best-characterized plant systems.as diploidyt It is a diploid plant permitting self pollination, and a relatively short generation time make it amenable to genetic analysis.

Classical genetics has created one of the largest stocks of morphological mutations induced by radiation (X-rays, UV-light, neutrons) and chemical mutagenesis. A major contributor in the mutagenesis area was Hans Stubbe who developed over 300 L. esculentum mutants and 200 in L. pimpinellifolium. (for summary see Rick, 1975). A particularly interesting example of induced mutagenesis was the directed manipulation of fruit size of L. esculentum and L. pimpinellifolium.(Stubbe, 1971).

The number of mapped genes in the form of cDNAs has increased considerably with the introduction of RFLP markers.

The current tomato map is considered to be complete in that all molecular and classical markers could be mapped to one of the 12 linkage groups indicating that no loci failed to link up with thethe map.

Propagation and Cultivation of Tomato [245]

Tomato is a highly specialized crop and bred to be grown under intensive monoculture. The distance required between foundation seed fields is 200 feet which in practical terms is the effective distance tomato pollen can travel under field conditions and remain viable.

Tomato Does Not Cross-pollinate Other Plant Species [245]

The factors that prevent cross-pollination are well documented and are applicable to genetically engineered tomato. Tomato can be crossed by hand-pollination to all wild Lycopersicon species with varying degrees of success. The genus has been divided into two subgenera, the one easily crossed with commercial tomato (esculentum complex), and those that cannot be easily crossed (peruvianum complex). Hybridization between these two subgenera usually leads to early embryo breakdown, which results in seed that is not viable. This problem can be circumvented by embryo culture and other laboratory techniques, albeit at great effort.

The closest genetic relatives of tomato are in the genus Solanum. Hybrids have been obtained between L. esculentum and S. lycopersicoides, but these hybrids are usually sterile (Stevens and Rick, 1986). No other member of the genus, including S. nigrum, a common weed in tomato fields, has yielded viable hybrids (Taylor, 1986).

There is no evidence that tomato plants can cross-pollinate with other plants in the area of the field test. Similarly, there is no evidence that the engineered tomato plants will cross-pollinate with any other tomato plants in the vicinity.

Dissemination of Transformed Tomato Plants [245]

The transformed tomato has gained no measurable selective advantage over nontransformed tomato in its ability to be disseminated or to become established in the environment.

The risk of horizontal movement of these genes from the genetically engineered tomato plants into the genetic environment is negligible because the genes are incorporated into the plant genome. No mechanism is known to exist in nature capable of transferring the genetic material from the tomato genome to another organism which tomato does not fertilize.

EU concept of coexistence [208]

The cultivation of genetically modified organisms (GMOs) in the EU is likely to have implications for the organisation of agricultural production. On the one hand, the possibility of the adventitious (unintended) presence of genetically modified (GM) crops in non-GM crops, and vice versa, raises the question as to how producer choice for the different production types can be ensured. In principle, farmers should be able to cultivate the types of agricultural crops they choose, be it GM crops, conventional or organic crops. None of these forms of agriculture should be excluded in the EU.

On the other hand, the issue is also linked to consumer choice. To provide European consumers with a real choice between GM food and non-GM food, there should not only be a traceability and labelling system that functions properly, but also an agricultural sector that can provide the different types of goods. The ability of the food industry to deliver a high degree of consumer choice goes hand in hand with the ability of the agricultural sector to maintain different production systems. Coexistence refers to the ability of farmers to make a practical choice between conventional, organic and GM-crop production, in compliance with the legal obligations for labelling and/or purity standards.

The adventitious presence of GMOs above the tolerance threshold set out in Community legislation triggers the need for a crop that was intended to be a non-GMO crop, to be labelled as containing GMOs. This could cause a loss of income, due to a lower market price of the crop or difficulties in selling it. Moreover, additional costs might incur to farmers if they have to adopt monitoring systems and measures to minimise the admixture of GM and non-GM crops. Coexistence is, therefore, concerned with the potential economic impact of the admixture of GM and non-GM crops, the identification of workable management measures to minimise admixture and the cost of these measures.

Foods derived from modern biotechnology Codex guidelines 2003

ftp://ftp.fao.org/codex/Publications/Booklets/Biotech/Biotech_2003e.pdf
The Codex Commission published risk analysis and food safety assessment of genetic modified foods in 2003 comprising:

1- Principles for the risk analysis of foods derived from modern Biotechnology.
2- Guideline for the conduct of food safety assessment of foods derived from recombinant-DNA plants.
3- Guideline for the conduct of food safety assessment of foods produced using recombinant-DNA microorganisms

While risk analysis has been used over a long period of time to address chemical hazards (e.g. residues of pesticides, contaminants, food additives and processing aids), and it is being increasingly used to address microbiological hazards and nutritional factors, the principles were not elaborated specifically for whole foods.

The risk analysis approach can, in general terms, be applied to foods including foods derived from modern biotechnology. However, it is recognised that this approach must be modified when applied to a whole food rather than to a discrete hazard that may be present in food.

The guidelines represent a risk analysis on the safety and nutritional aspects of foods derived from modern biotechnology. Environmental, ethical, moral and socio-economic aspects of the research, development, production and marketing of these foods are not covered.

Genetically engineered food labelling, the position of FDA [246]

Statements about foods developed using bioengineering

FDA recognizes that some manufacturers may want to use informative statements on labels and in labeling of bioengineered foods or foods that contain ingredients produced from bioengineered foods. The following are examples of some statements that might be used. The discussion accompanying each example is intended to provide guidance as to how similar statements can be made without being misleading.

"Genetically engineered" or "This product contains cornmeal that was produced using biotechnology."

The information that the food was bioengineered is optional and this kind of simple statement is not likely to be misleading. However, focus group data indicate that consumers would prefer label statements that disclose and explain the goal of the technology (why it was used or what it does for/to the food). Consumers also expressed some preference for the term "biotechnology" over such terms as "genetic modification" and "genetic engineering"

. "This product contains high oleic acid soybean oil from soybeans developed using biotechnology to decrease the amount of saturated fat."

This example includes both required and optional information. As discussed above in the background section, when a food differs from its traditional counterpart such that the common or usual name no longer adequately describes the new food, the name must be changed to describe the difference. Because this soybean oil contains more oleic acid than traditional soybean oil, the term "soybean oil" no longer adequately describes the nature of the food. Under section 403(i) of the act, a phrase like "high oleic acid" would be required to appear as part of the name of the food to describe its basic nature. The statement that the soybeans were developed using biotechnology is optional. So is the statement that the reason for the change in the soybeans was to reduce saturated fat.

"These tomatoes were genetically engineered to improve texture."

In this example, the change in texture is a difference that may have to be described on the label. If the texture improvement makes a significant difference in the finished product, sections 201(n) and 403(a)(1) of the act would require disclosure of the difference for the consumer. However, the statement must not be misleading. The phrase "to improve texture" could be misleading if the texture difference is not noticeable to the consumer. For example, if a manufacturer wanted to describe a difference in a food that the consumer would not notice when purchasing or consuming the product, the manufacturer should phrase the statements so that the consumer can understand the significance of the difference. If the change in the tomatoes was intended to facilitate processing but did not make a noticeable difference in the processed consumer product, a phrase like "to improve texture for processing" rather than "to improve texture" should be used to ensure that the consumer is not misled. The statement that the tomatoes were genetically engineered is optional.

"Some of our growers plant tomato seeds that were developed through biotechnology to increase crop yield."

The entire statement in this example is optional information. The fact that there was increased yield does not affect the characteristics of the food and is therefore not necessary on the label to adequately describe the food for the consumer. A phrase like "to increase yield" should only be included where there is substantiation that there is in fact the stated difference.

Where a benefit from a bioengineered ingredient in a multi-ingredient food is described, the statement should be worded so that it addresses the ingredient and not the food as a whole; for example, "This product contains high oleic acid soybean oil from soybeans produced through biotechnology to decrease the level of saturated fat." In addition, the amount of the bioengineered ingredient in the food may be relevant to whether the statement is misleading. This would apply especially where the bioengineered difference is a nutritional improvement. For example, it would likely be misleading to make a statement about a nutritionally improved ingredient on a food that contains only a small amount of the ingredient, such that the food's overall nutritional quality would not be significantly improved.

FDA reminds manufacturers that the optional terms that describe an ingredient of a multi-ingredient food as bioengineered should not be used in the ingredient list of the multi-ingredient food. Section 403(i)(2) of the act requires each ingredient to be declared in the ingredient statement by its common or usual name. Thus, any terms not part of the name of the ingredient are not permitted in the ingredient statement. In addition, 21 CFR 101.2(e) requires that the ingredient list and certain other mandatory information appear in one place without other intervening material. FDA has long interpreted any optional description of ingredients in the ingredient statement to be intervening material that violates this regulation.

Statements about foods that are not bioengineered or that do not contain ingredients produced from bioengineered foods [246]

Terms that are frequently mentioned in discussions about labeling foods with respect to bioengineering include "GMO free" and "GM free." "GMO" is an acronym for "genetically modified organism" and "GM" means "genetically modified." Consumer focus group data indicate that consumers do not understand the acronyms "GMO" and " GM" and prefer label statements with spelled out words that mean bioengineering.

Terms like "not genetically modified" and "GMO free," that include the word "modified" are not technically accurate unless they are clearly in a context that refers to bioengineering technology. "Genetic modification" means the alteration of the genotype of a plant using any technique, new or traditional. "Modification" has a broad context that means the alteration in the composition of food that results from adding, deleting, or changing hereditary traits, irrespective of the method.

Modifications may be minor, such as a single mutation that affects one gene, or major alterations of genetic material that affect many genes. Most, if not all, cultivated food crops have been genetically modified. Data indicate that consumers do not have a good understanding that essentially all food crops have been genetically modified and that bioengineering technology is only one of a number of technologies used to genetically modify crops. Thus, while it is accurate to say that a bioengineered food was "genetically modified," it likely would be inaccurate to state that a food that had not been produced using biotechnology was "not genetically modified" without clearly providing a context so that the consumer can understand that the statement applies to bioengineering.

The term "GMO free" may be misleading on most foods, because most foods do not contain organisms (seeds and foods like yogurt that contain microorganisms are exceptions). It would likely be misleading to suggest that a food that ordinarily would not contain entire "organisms" is "organism free."

There is potential for the term "free" in a claim for absence of bioengineering to be inaccurate. Consumers assume that "free" of bioengineered material means that "zero" bioengineered material is present. Because of the potential for adventitious presence of bioengineered material, it may be necessary to conclude that the accuracy of the term "free" can only be ensured when there is a definition or threshold above which the term could not be used. FDA does not have information with which to establish a threshold level of bioengineered constituents or ingredients in foods for the statement "free of bioengineered material."

FDA recognizes that there are analytical methods capable of detecting low levels of some bioengineered materials in some foods, but a threshold would require methods to test for a wide range of genetic changes at very low levels in a wide variety of foods. Such test methods are not available at this time. The agency suggests that the term "free" either not be used in bioengineering label statements or that it be in a context that makes clear that a zero level of bioengineered material is not implied. However, statements that the food or its ingredients, as appropriate, was not developed using bioengineering would avoid or minimize such implications. For example,
- "We do not use ingredients that were produced using biotechnology;"
- "This oil is made from soybeans that were not genetically engineered;" or
- "Our tomato growers do not plant seeds developed using biotechnology."

A statement that a food was not bioengineered or does not contain bioengineered ingredients may be misleading if it implies that the labeled food is superior to foods that are not so labeled. FDA has concluded that the use or absence of use of bioengineering in the production of a food or ingredient does not, in and of itself, mean that there is a material difference in the food. Therefore, a label statement that expresses or implies that a food is superior (e.g., safer or of higher quality) because it is not bioengineered would be misleading. The agency will evaluate the entire label and labeling in determining whether a label statement is in a context that implies that the food is superior.

In addition, a statement that an ingredient was not bioengineered could be misleading if there is another ingredient in the food that was bioengineered. The claim must not misrepresent the absence of bioengineered material. For example, on a product made largely of bioengineered corn flour and a small amount of soybean oil, a claim that the product "does not include genetically engineered soybean oil" could be misleading.

Even if the statement is true, it is likely to be misleading if consumers believe that the entire product or a larger portion of it than is actually the case is free of bioengineered material. It may be necessary to carefully qualify the statement in order to ensure that consumers understand its significance.

Further, a statement may be misleading if it suggests that a food or ingredient itself is not bioengineered, when there are no marketed bioengineered varieties of that category of foods or ingredients. For example, it would be misleading to state "not produced through biotechnology" on the label of green beans, when there are no marketed bioengineered green beans.

To not be misleading, the claim should be in a context that applies to the food type instead of the individual manufacturer's product. For example, the statement "green beans are not produced using biotechnology" would not imply that this manufacturer's product is different from other green beans.

Substantiation of label statements [246]

A manufacturer who claims that a food or its ingredients, including foods such as raw agricultural commodities, is not bioengineered should be able to substantiate that the claim is truthful and not misleading. Validated testing, if available, is the most reliable way to identify bioengineered foods or food ingredients. For many foods, however, particularly for highly processed foods such as oils, it may be difficult to differentiate by validated analytical methods between bioengineered foods and food ingredients and those obtained using traditional breeding methods.

Where tests have been validated and shown to be reliable they may be used. However, if validated test methods are not available or reliable because of the way foods are produced or processed, it may be important to document the source of such foods differently. Also, special handling may be appropriate to maintain segregation of bioengineered and nonbioengineered foods.

In addition, manufacturers should consider appropriate recordkeeping to document the segregation procedures to ensure that the food's labeling is not false or misleading. In some situations, certifications or affidavits from farmers, processors, and others in the food production and distribution chain may be adequate to document that foods are obtained from the use of traditional methods. A statement that a food is "free" of bioengineered material may be difficult to substantiate without testing. Because appropriately validated testing methods are not currently available for many foods, it is likely that it would be easier to document handling practices and procedures to substantiate a claim about how the food was processed than to substantiate a "free" claim.

FDA has been asked about the ability of organic foods to bear label statements to the effect that the food (or its ingredients) was not produced using biotechnology. On December 21, 2000, the Agriculture Marketing Service of the U.S. Department of Agriculture (USDA) published final regulations on procedures for organic food production (National Organic Program final rule; 65 FR 80548). That final rule requires that all but the smallest organic operations be certified by a USDA accredited agent and lays out the requirements for organic food production.

Among those requirements is that products or ingredients identified as organic must not be produced using biotechnology methods. The national organic standards would provide for adequate segregation of the food throughout distribution to assure that non-organic foods do not become mixed with organic foods. The agency believes that the practices and record keeping that substantiate the "certified organic" statement would be sufficient to substantiate a claim that a food was not produced using bioengineering.

Genetic regulations in the EU

EU Release directive 2001/18EG

Directive 2001/18/EC of the European Parliament and of the Council of 12 March 2001 on the deliberate release into the environment of genetically modified organisms and repealing Council Directive 90/220/EEC - Commission Declaration.
http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2001:106:0001:0038:EN:PDF

Genetically modified food and feed directive (EG) 1829/2003[194]

Regulation (EC) No 1829/2003 of the European Parliament and of the Council of 22 September 2003 on genetically modified food and feed regulates labelling og GM food. The article 12 (1) establishes a labelling threshold of 0.9% in European Union (EU). This threshold is set to 3% in Korea, and 5% in Japan. In China there is zero tollerance for labelling of 17 types of GM products. This includes maize seeds, maize oil, tomato seeds, ketchup, soybean seeds, soybean oil, rapeseed seeds, and cotton seeds. [247]

This considered the almost worldwide GMO comtamination of seeds, tranportation,storage facilities and production lines.

Gene register decision

Commission Decision 2004/204/EC of 23 February 2004 laying down detailed arrangements for the operation of the registers for recording information on genetic modifications in GMOs, provided for in Directive 2001/18/EC of the European Parliament and of the Council.
http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2004:065:0020:0022:EN:PDF

EFSA-GMO Nr. Event/Species Scope of the
    application
UK-2004-1 NK603 x MON810 Maize Food, Feed
NL-2004-02 1507 Maize Food
DE-2004-03 MON863 x MON810 Maize Food, feed
UK-2004-04 Rice LLRICE62 Food,feed
    import and processing
UK-2004-05 1507 x NK603 Maize Food, feed
    import and processing
UK-2004-06 MON863 x NK603 Maize Food, feed
    Import and processing
BE-2004-07 MON863 x MON810 x Food, feed
  NK603 Maize import and processing
UK-2004-08 H7-1 Sugar beet Food, feed produced
    from GM plants
    (derived products)
UK-2005-09 MON 531 x MON 1445 Food, feed produced
  Cotton from GM plants
    (derived products)
  MON 15985 and Food, feed produced
UK-2005-10 MON 15985 x MON from GM plants
  1445 Cotton (derived products)
UK-2005-11 MIR604 Maize Food, feed
    Import and processing
NL-2005-12 591223 Maize Food, feed
    Import and processing
NL-2005-13 LLCotton25 Fodd,feed??
    Import and processing
UK-2005-14 Amylopectin Potato Food, feed
  event EH92-527-1  
NL-2005-15 1507 x 59122 Maize Food, feed
    Import and processing
NL-2005-16 281-24-236 x Food, feed
  3006-210-23 Cotton Food, feed
UK-2005-17 1507 x NK603 Maize Food, feed, Import
    and processing,
    CULTIVATION
NL-2005-18 A2704-12 Soybean Food, feed
    Import and processing
UK-2005-19 GA21 Maize Food, feed
    Import and processing
UK-2005-20 59122 x NK603 Maize Food, feed
    Import and processing
UK-2005-21 59122 x 1507 x NK603 Food, feed
    Import and processing
NL-2005-22 NK603 Maize Food, feed
    Import and processing
    CULTIVATION
NL-2005-23 59122 Maize Food, feed
    Import and processing
    CULTIVATION
NL-2005-24 40-3-2 Soybean CULTIVATION
UK-2005-25 T45 Oilseed rape Food, feed
    Import and processing
NL-2005-26 NK603 x MON810 Maize CULTIVATION
CZ-2005-27 MON 88017 Maize Food, feed
    Import and processing
NL-2005-28 1507 x 59122 Maize Food, feed
    Import and processing
    CULTIVATION
UK-2006-29 59122 x NK603 Maize Food, feed
    Import and processing
    CULTIVATION
UK-2006-30 59122 x 1507 x NK603 Food, feed
    Import and processing
    CULTIVATION
NL-2006-31 LY038 Maize Food, feed
    Import and processing
NL-2006-32 LY038 x MON810 Maize Food, feed
    Import and processing
CZ-2006-33 MON 88017 x MON 810 Maize Food, feed
    Import and processing
UK-2006-34 Event 3272 Maize Food, feed
    Import and processing
NL-2006-35 LLCotton25 x MON Food,feed produced
  15985 Cotton from GM plants
    (derived producs)



Glyphosate-tolerant genetically modified maize GA21 from Syngenta [248]

The GMO Maize GA21 applyed for food and feed uses, import and processing and for renewal of the authorisation of maize GA21.

The GMO Panel of EFSAconsiders maize GA21, on October 2007, as safe as its non genetically modified counterparts with respect to potential effects on human and animal health or the environment. Therefore the GMO Panel concludes that maize GA21 is unlikely to have any adverse effect on human and animal health or on the environment in the context of its intended uses.

Insect-resistant GMO Maize 59122 approved in EU [249]

The maize 59122 from Pioneer Hi-Bred International, Inc. and Mycogen Seeds, c/o Dow Agrosciences LLC was approved on the 23 March 2007 by the Panel of the European Food Safety Authority for food and feed uses, import and processing of maize 59122 but does not include cultivation.

Maize 59122 was transformed by Agrobacterium-mediated gene transfer technology and expresses CRY34Ab1, CRY35Ab1 and PAT proteins. The molecular characterisation data established that maize 59122 contains a single insert of the T-DNA. The structure of the insert in maize 59122 was determined by Southern analysis and DNA sequencing. No vector backbone sequences were detected. BLAST sequence analysis revealed that border regions of the maize event 59122 show significant homology to maize genomic DNA and EST sequences. None of the EST sequences showed homology to known toxin or allergen encoding sequences. Analysis of ORFs spanning the two junction regions was performed by bioinformatic analysis and no novel ORFs with sequence similarity to known toxins or allergens were identified.

the GMO Panel concludes that maize 59122 is unlikely to have any adverse effect on human and animal health or on the environment in the context of its intended uses.

GM maize Bt11xGA21 from Synngenta Seeds is safe [250]

The EFSA GMO Panel on the 17.09.2009 considers that maize Bt11xGA21 is as safe as its non-GM maize counterpart with respect to effects on human and animal health and the environment. Therefore, the EFSA GMO Panel concludes that maize Bt11xGA21 is unlikely to have any adverse effect on human and animal health or on the environment in the context of its intended uses.

GM corn in Europe

Dow AgroSciences is engaged to develop genetic solutions for crop production for food and non-food agriculture. With the acquisition Austrian Maize Technologies, and the Dutch germplasm company Duo Maize, Dow AgroSiences will become leading in the European market of corn seeds for grain and silage.

Herculex [251]

The new acquisition will foster the spread of the Herculex seed in the European market which covers about 12 million hectares of corn fields.

The European registration of Dow's Herculex corn which is expected for soon, will bring more protection of corn against more destructive pests than any other trait, according to DowAgroSciences. It is the only in-plant technology that protects against western bean cutworm and black cutworm.

Full package of GM technology

HERCULEX expresses the Cry 1F Bt protein and provides protection against above-ground insects like European corn borer, western bean cutworm, black cutworm and fall armyworm.
It combines Cry 1F with the binary Bt proteins, Cry 34Ab1 and Cry 35Ab1 to control all listed pests plus corn rootworm in HERCULEX XTRA and Pioneer Hi-Bred. Herculex traits also contain LibertyLink technology and many Herculex hybrids contain Roundup Ready Corn 2 technology.

Dow agroSciences presents three main traits:
Herculex I: Dow says its Herculex I covers in one stroke the followingh pests:
European corn borer
Southwestern corn borer
Western bean cutworm
Black cutworm
Fall Armyworm
Southern corn stalk borer
Lesser cornstalk borer
Sugarcane borer
Corn earworm suppression

Herculex RW Rootworm Protection

provides powerful, below-ground in-plant insect protection against western, northern and Mexican corn rootworm larvae, during the whole season.

Herculex XTRA

combines Herculex I Insect Protection and Herculex RW Rootworm Protection for powerful protection above-and below-ground. Its whole-plant protection works all season long and shields your corn unlike any other trait.

A perfect family of seed, pesticides and herbicides

Adopting Herculex the door is open to a large family of GM technologies where everything is well round up. However, Dow AgroSciences warns: Application of other, not specified herbicides to Herculex corn could result in total crop loss. The use of Roundup Ready Corn 2 technology requires the purchaser to have completed a Monsanto Technology Agreement.

Bayer CropScience [252]

Bayer CropScience has developed partnerships with each of the participating seed companies listed below to offer LIBERTY herbicide at a better value by reducing the LIBERTY herbicide cost. See the list of corn seeds which have the Liberty Link gene at
http://www.bayergrowingstrong.com/liberty_rebate/

Genetically Engineered Enzymes [253]

Enzymes are naturally occurring proteins that speed up biochemical processes. They're used to produce everything from wine and cheese to corn syrup and baked goods.

When enzymes are genetically engineered, the manufacturer is not required to notify the FDA that the enzymes have been modified. The FDA provides an incomplete list of GE enzymes:

[253]
Enzymes Used for
Chymosin production of cheese
Novamyl(TM) production of baked goods, preserves freshness
Alpha amylase white sugar, maltodextrins and nutritive
  carbohydrate sweeteners (corn syrup)
Aspartic (proteinase enzyme from R. miehei) production of cheese
Pullulanase production of high fructose corn syrup

FDA Food safety evaluation of new Non-Pesticidal proteins [244]

The FDA asserts in its policy on GM foods that genetically engineered crops are no different than those created through traditional breeding methods. The agency bases its position on the fact that foods derived from traditionally bred crops have a history of safety. Thus the FDA takes the position that genetic engineering is just another traditional breeding method, and reasons that GM foods should be considered safe.

The Food and Drug Administration (FDA) issued a final guidance for industry entitled "Recommendations for the Early Food Safety Evaluation of New Non- Pesticidal Proteins Produced by New Plant Varieties Intended for Food Use." The guidance provides recommendations to developers of new plant varieties, including bioengineered plant varieties, on the early food safety evaluation of new non-pesticidal proteins produced by such new plant varieties, including for example such proteins produced in bioengineered plants. The guidance describes procedures for submitting an early food safety evaluation of such proteins to the agency.

The guidance describes the procedure for early food safety evaluation of new proteins in new plant varieties that are under development for food use. In most cases, the proteins expected to become components of food, whether as a result of the use of traditional or modern biotechnology methods, will be the same or quite similar to proteins commonly found in food. FDA believes that any food safety concern related to such material entering the food supply would be limited to the potential that a new protein in food from the plant variety could cause an allergic reaction in susceptible people or could be a toxin in people or animals.

FDA recommends that sponsors and developers of new plant varieties intended for food use consult with FDA about their evaluation of the food safety of any new proteins produced in these plants prior to the stage of development where the new proteins might inadvertently enter the food supply. Thus, the safety evaluation recommended by this guidance is termed an "early" food safety evaluation of new proteins.

If a protein has been evaluated in an early food safety evaluation and no safety concerns are identified, we would not expect an additional early food safety evaluation to be submitted if the same protein is introduced into another plant species. Also, if a protein has previously been reviewed as part of a biotechnology consultation and there were no safety concerns identified, we would not expect you to submit an early food safety evaluation for such a protein. This guidance does not apply to plant-incorporated protectants (PIPs), which are regulated by EPA.

The Environmental Protection Agency (EPA) is responsible for evaluating the safety of pesticides, including plant-incorporated protectants. As such, these proteins are not subject to FDA review and are not the subject of this guidance.

Tomato chloroplast DNA transformation as promising nutraceuticals [254]

Genetic engineering scientist are searching the way of production of food crops with enhanced nutritional or medicinal value, the so called nutraceuticals.

Ralf Bock and colleagues from the Institute of Plant Biochemistry and Biotechnology in Münster, Germany, in cooperation with Helaine Carrer of the Centre of Agriculture Biotechnology of Piracicaba, Brazil, created a new technique of genetic engineering. Instead of modifying the DNA of the nucleus of the cell, the scientists modified the DNA of plastids such as chloroplasts. In this way efficient production of edible vaccines, pharmaceuticals, and antibodies in tomato can be realised, while the new genes are hindered to pass to other crops or weeds.

This technique had already been used in genetic engineering of tobacco. Ralf Bock and his team optimized plant tissue culture and regeneration, nutrition, plant hormones and light intensity for the tomato plant.

Only one marker gene has been used yet, but more genes will now be focused in the experiments of Ralf Bock.

The idea to use GM tomatoes with transformed plastids to develop edible vaccines is also backed up by Pal Maliga from the University in Piscataway, New Jersey. [255]

Some of the technical issues that require resolution comprising the transfer of foreign genes from chloroplasts to the nucleus were described by Henry Daniell and Christopher L. Parkinson, University of Central Florida. [256]

Antibiotic Resistance Marker Genes and approval of APH(3')II gene [257]

The FDA position to antibiotic resistance marker genes is given in the draft guidance "Use of Antibiotic Resistance Marker Genes in Transgenic Plants: Guidance for Industry" (the draft guidance) and report entitled "Report on Consultations Regarding Use of Antibiotic Resistance Marker Genes In Transgenic Plants."

The report summarizes FDA's recent consultations with outside experts on the use of antibiotic resistance marker genes in transgenic plants. The draft guidance is intended to provide information to crop developers that will assist them on the use of antibiotic resistance marker genes in the development of transgenic plants.

In the Federal Register of May 29, 1992 (57 FR 22984), FDA published a notice on a policy statement (the 1992 policy) regarding foods derived from new plant varieties, including those derived using genetic engineering techniques. In the 1992 policy statement, FDA specifically discussed antibiotic resistance selectable marker genes and noted that both the antibiotic resistance gene and the enzyme encoded by the gene, unless removed, are expected to be present in foods derived from plants developed using the markers.

The agency acknowledged that, when present in food, enzymes that are encoded by selectable marker genes and that inactivate certain clinically useful antibiotics theoretically might reduce the therapeutic efficacy of antibiotics administered orally. Accordingly, FDA believes that it is important to evaluate such concerns with respect to commercial use of antibiotic resistance marker genes in food, especially those marker genes that will be widely used. In addition, the agency also believes that it is important to consider the possibility that resistance to antibiotics in microorganisms has the potential to spread through horizontal transfer of antibiotic resistance marker genes from plants (59 FR 26700, May 23, 1994).

This second consideration was reflected in FDA's evaluation of the safety of the use of the kanamycin resistance (kan<SUP>r</SUP>) gene product, aminoglycoside 3'-phosphotransferase II (APH(3')II, also known as neomycin phosphotransferase II or nptII) when the agency amended the food additive regulations to permit the use of APH(3')II in the development of transgenic tomato, cotton, and oilseed rape.

FDA received several comments from the public regarding the use of antibiotic resistance marker genes in transgenic plants in response to the 1992 policy, and in response to the agency's solicitation for comment regarding the request of Calgene, Inc., for an advisory opinion on the use of the kan<SUP>r</SUP> gene as a selectable marker in the development of transgenic tomatoes (56 FR 20004, May 1, 1991). FDA responded to these comments when it issued the final rule permitting the use of APH(3')II in the development of transgenic tomatoes, oilseed rape, and cotton (59 FR 26700 at 26706).

Since FDA's decision approving the use of the APH(3')II in the development of transgenic tomatoes, cotton, and oilseed rape, the agency has continued to receive inquiries from crop developers as well as from the public regarding the safety and regulatory status of antibiotic resistance marker genes. Therefore, FDA sought to develop sound scientific principles regarding the safety of the use of antibiotic resistance marker genes in the development of transgenic plants intended for food use so as to provide sound scientific guidance to crop developers regarding the safe use of antibiotic resistance marker genes.

Toward this end, FDA undertook several consultations with outside experts having expertise in relevant fields including gene transfer and antibiotic resistance. The purpose of the consultations was to determine whether circumstances exist under which FDA should recommend that a given antibiotic resistance gene not be used in crops intended for food use, and if so, to delineate the nature of those circumstances.

Guidance for Industry: Use of Antibiotic Resistance Marker Genes in Transgenic PlantsDraft Guidance. September 4, 1998 [211]

In 1992, the Food and Drug Administration (FDA) issued a policy statement regarding foods derived from new plant varieties including those derived using genetic engineering techniques (U.S. FDA, 1992). In this policy statement, FDA specifically discussed antibiotic resistance selectable marker genes and noted that both the antibiotic resistance gene and gene product, unless removed, are expected to be present in foods derived from plants developed using the markers. The agency acknowledged that selectable marker gene-encoded enzymes that inactivate certain clinically useful antibiotics, when present in food, theoretically might reduce the therapeutic efficacy of antibiotics administered orally.

Thus it is important to evaluate such concerns with respect to commercial use of antibiotic resistance marker genes in food, especially those that will be widely used. In addition, the agency believes it is important to consider the possibility that resistance to antibiotics in microorganisms might spread through potential horizontal transfer of antibiotic resistance marker genes from plants to microorganisms in the gastrointestinal tract or in the environment.

Since FDA's decision regarding the use of the kanamycin resistance (kanr) gene product, aminoglycoside 3'-phosphotransferase II (APH(3')II, also known as neomycin phosphotransferase II or nptII) in the development of transgenic tomato, cotton, and oilseed rape,

the agency has continued to receive inquiries regarding the safety and regulatory status of antibiotic resistance marker genes. Therefore, FDA sought to develop sound scientific principles regarding the safety of the use of antibiotic resistance marker genes in the development of transgenic plants for food use and to provide sound scientific guidance to crop developers regarding the safe use of antibiotic resistance marker genes.

Towards this end, FDA undertook several consultations with outside experts between November, 1996 and February, 1997. The purpose of the consultations was to determine whether circumstances exist under which FDA should recommend that a given antibiotic resistance gene not be used in crops intended for food use, and if so, to delineate the nature of those circumstances.

Overall, the arguments made concerning the improbability of the transfer of the kanr gene to gut microorganisms in the case of the Flavr Savr tomatoes (U.S. FDA, 1994) could also be applied to other antibiotic resistance marker genes. However, while the possibility of transfer from plants to microorganisms is remote for all marker genes, crop developers should consider the following factors in evaluating whether an antibiotic resistant gene is suitable for use as a selectable marker:
1) whether the antibiotic that may be affected is clinically important,
2) whether it is frequently used,
3) whether it is administered orally,
4) whether it is unique or acceptable alternative antibiotics exist,
5) whether there would be selective pressure for transformants to be selected, and
6) whether there already is resistance to the antibiotic in the environment.

FDA's analysis (U.S. FDA, 1994) showed that the kanr gene that was used in the case of the Flavr Savr tomato passes the paradigm outlined above. Neomycin and kanamycin are infrequently used antibiotics, neither is unique for any use, and rarely are administered orally. Thus, selective pressure would be minimal for development of resistant bacteria because the drugs are not used in humans or in animals to any great extent. Similarly, these antibiotics are not used in agriculture or aquaculture to any great extent. Therefore, they would not provide the selective pressure required to select for the kanr determinant if transfer from plants to soil microorganisms were to take place. In addition, existing resistance levels far exceed any transfer that may take place from transgenic plants to microorganisms.

However, different circumstances may apply to other antibiotics. For example, with regard to the presence or absence of selective pressure, streptomycin and oxytetracycline may provide selective pressure in the environment because of their use as pesticides in agriculture. On the other hand, ampicillin may provide selective pressure in the human gut when used in the clinical setting but not in the environment.

The notion that it may be possible to construct a list of antibiotic resistance marker genes that are acceptable for use in the development of transgenic crops was discussed. The kanr gene can be placed on such a list. Some experts suggested that the hygromycin resistance gene may be included on such a list because of its limited use in humans. It was noted, however, that it may have important veterinary uses.

Other experts would include the beta-lactamase gene of pUC18 (that confers resistance to a narrow spectrum of beta-lactam antibiotics), and the tetracycline resistance gene on such a list. There is so much resistance to these antibiotics already in the environment that any potential transfer from transgenic plants to microorganisms, especially when compared to transfer among bacteria, is unlikely to add to the existing levels of resistance in any meaningful way.

The idea was put forth that antibiotic resistance genes could be ranked on a continuum with the kanr gene on one end as the most acceptable, and the vancomycin resistance gene on the other. It was further suggested that use of marker genes beyond the kanr and hygromycin resistance genes might be acceptable on the basis of studies to address potential transfer and a commitment to conduct post-market surveillance for transfer of the gene in question.

Potential Transfer of Antibiotic Resistance Marker Genes to Microroganism in the Environment [211]

Some experts noted that some soil microbes may be naturally transformable and that they may take up and incorporate DNA causing genomic rearrangements that might help them occupy particular ecological niches.

Some experts felt that it does not make sense to expand the availability of the resistance gene in the environment because abundance of marker genes may compress the typical 4- 5-year time lag between first use of new antibiotic and the emergence of resistance in hospitals.

Others felt that the risk of transfer from plant genome to soil microorganisms is not a significant one. This latter group felt that DNA from plant debris would be unavailable for transfer because it would be degraded by nucleases when the plant cell lysed; in addition, there would be no selective pressure in most cases although there are exceptions such as when streptomycin and tetracycline are used as pesticides to prevent fire blight in fruit trees, or when manure is used as fertilizer following use of antibiotics as growth promoters in animals. It was noted that transfer from bacteria to bacteria accounts for the wide dissemination of certain antibiotic resistance markers in soil bacteria.

Approaches to Assessing Potential for Transfer of Antibiotic Resistance Marker Genes and Conducting Surveillance for Resistance [211]

Participants in the consultations discussed a study conducted by a crop developer wherein plant DNA containing the beta-lactamase gene from an insect-resistant transgenic corn line (intact or nuclease degraded) was incubated with competent E. coli. The study was intended to determine if any bacteria were transformed and acquired ampicillin resistance; the experiment showed that transformation did not occur above a frequency of 1 in 6.8 x1019.

Some experts said that if transformation were to take place, it would be more likely to do so in experiments using competent bacteria in the laboratory than in nature because competent bacteria have the highest transformation frequency. They added that if transformation was not observed in the laboratory, especially if the experiment was carried out in gram negative and gram positive bacteria (an Enterococcus strain and an E. coli), the results would suggest that such transfers may not take place in the natural setting to the extent that they would raise health or safety concerns.

Other experts stated that an in vitro experiment does not give them much confidence because it does not reflect the complex ecological system that exists in nature. In addition, a monoculture of E. coli, is an artificial system that would not be a strong basis on which to assess risk.

Conclusions [211]

German executiv agencys contact for genetic engineering

The Bavarian Environmental Agency (Bayerisches Amt für Umweltschutz) http://www.bayern.de/lfu/gentec/index.html provides useful service for genetic engineering projects, the registration, the release of genetic modified organism, to place a GMO on the market, the official supervision of fields, texts of relevant regulations, a link to the Federal Agency for Consumer Protection and Food Safety (Bundesamt für Verbraucherschutz und Lebensmittelsicherheit (BVL) ) is provided. http://www.bvl.bund.de/cln_027/nn_495478/DE/06__Gentechnik/gentechnik__node.html__nnn=true The Federal Agency for Consumer Protection and Food Safety is responsible for the entire genetic legislation enforcement since 2004.

Tetracycline residues in milk [258]

Tetracycline

is a broad-spectrum polyketide antibiotic produced by the Streptomyces bacterium, used to treat bacterial infections, including pneumonia and other respiratory tract infections; acne; infections of skin, genital and urinary systems; and the infection that causes stomach ulcers (Helicobacter pylori). It also may be used as an alternative to other medications for the treatment of Lyme disease and for the treatment and prevention of anthrax (after inhalational exposure). Tetracycline is in a class of medications called tetracycline antibiotics.

Tetracyclines

are a group of broad-spectrum antibiotics whose general usefulness has been reduced with the onset of bacterial resistance. Despite this, they remain the treatment of choice for some specific indications. They are so named for their four ("tetra-") hydrocarbon rings. More specifically, they are defined as a subclass of polyketides having an octahydrotetracene-2-carboxamide skeleton. They are collectively known as derivatives of polycyclic naphthacene carboxamide.
Image Tetracycline Source:Wikipedia.

Mecanism and resistance [259]

Tetracycline inhibits cell growth by inhibiting translation. It binds to the 16S part of the 30S ribosomal subunit and prevents the amino-acyl tRNA from binding to the A site of the ribosome. The binding is reversible in nature.

Cells become resistant to tetracycline by at least three mechanisms: enzymatic inactivation of tetracycline, efflux, and ribosomal protection. Inactivation is the rarest type of resistance, where an acetyl group is added to the molecule, causing inactivation of the drug. In efflux, a resistance gene encodes a membrane protein that actively pumps tetracycline out of the cell. This is the mechanism of action of the tetracycline resistance gene on the artificial plasmid pBR322. In ribosomal protection a resistance gene encodes a protein which can have several effects depending on what gene is transferred. Six classes of ribosomal protection genes/proteins have been found, all with high sequence homology suggesting a common evolutionary ancestor.

Possible mechanisms of action of these protective proteins include: 1.blocking tetracyclines from binding to the ribosome, 2.binding to the ribosome and distorting the structure to still allow t-RNA binding while tetracycline is bound, and 3.binding to the ribosome and dislodging tetracycline.

Veterinary use of tetracyclines [260]

The tetracyclines group of dairy antibiotics is used to treat bovine mastitis in dairy herds, and includes tetracycline, chlortetracycline, and oxytetracycline. Quarantine of treated animals up to 4 days is necessary to avoid tetracycline residues higher than 100 parts per billion in milk and their products, established by the European Union. Tetracyclines are widely used in Spain and other European countries.

Dip-stick test for tetracyclines in milk

A new dip-stick test from Neogen's TetraStar(R) monitoring tetracyclines in milk charges, gives results in six minutes. Another test is necessary to control the beta-lactam group of dairy antibiotics, which includes amoxicillin, ampicillin, cephapirin, cloxacillin, and penicillin. This group is widely used throughout the world in dairy veterinary medication.


Gram-negative Enterobacteriaceae with resistance to carbapenem in multidrug-resistant bacteria [261]

The Lancet warn of a pandemic of antibiotic-resistant strains can easily be spread by international travel or during gatherings such as Hajj, turning containment of microbial resistance imperious. The warning is based on the study of Kumarasamy et al.

The epidemiological study of Kumarasamy et al. 2010 identified 180 isolates, collected in India, Pakistan and UK. the presence of the carbapenem resistance gene blaNDM-1 was established NDM-1 was mostly found among Escherichia coli and Klebsiella pneumoniae, which were highly resistant to all antibiotics except to tigecycline and colistin. The authors concluded that the New Delhi metallo-beta-lactamase 1 (NDM-1) strains are a global threat and call for a co-ordinated international surveillance. [262]


Increasing prevalence of NDM-1 ertapenem non-susceptible isolates in India [263]

Ertapenem is a beta-lactam reserve antibiotic. Ertapenem-non-susceptible isolates of Enterobacteriaceae from the worldwide Study for Monitoring Antimicrobial Resistance Trends (SMART) 2009 programme were screened by Lascols et al 2011.

The authors found strains containing the carbapenemase genes bla(NDM-1), both bla(NDM-1) and bla(OXA-48); other carbapenemase genes included bla(KPC), bla(VIM) and bla(OXA-48). All bla(NDM-1)-carrying isolates were from patients in India

The authors concluded that carbapenemase genes among Enterobacteriaceae are associated with diverse ESBLs and/or AmpC backgrounds, and carried various combinations of beta-lactamases.

Other multidrug-resistant strains [264]

Community-acquired methicillin-resistant Staphylococcus aureus, or ca-MRSA are resistant to almost all common antibiotics. The drug-resistant hospital-acquired MRSA (ha-MRSA) strains affect elderly and people in hospitals and nursing homes, ca-MRSA strain affects healthy young people.

In October 2010 Friends of the earth stressed that antibiotics, hormones and other drugs are used to turn factory farming profitable. The organisation suggests alternatives. Factory farming is a method of farming where livestock is raised in confinement in large numbers (and at high density) in a factory-like environment. For the system to work, it needs high volumes of cheap animal feed as well as antibiotics and pesticides to mitigate the spread of disease exacerbated by the crowded living conditions. [265]


ESBL bacteria in Germany [266]

The German Friends of the Earth (BUND) found multidrug-resistant bacteria on more than half of the chicken parts purchased in supermarkets. BUND also found MRSA bacteria on one of Germany's high-speed ICE trains, German retirement home, on beef, pork and vegetables. Alarming finding are symptom-free carries of ESBL-forming bacteria in the intestine. Such bacteria killed neonatal at a hospital in the northern German city of Bremen was infested with an ESBL-forming bacterium last fall.

Some pathogens already developed resistance to drugs of last resort. In India, where poor hygiene and the availability of over-the-counter antibiotics encourage the development of resistance. Infected with these types of pathogens die of urinary tract infections, wound infections or pneumonia. The killer bugs contaminated travellers and infection cases were reported in England, Germany and Israel.

Some 900 metric tons of antibiotics are administered to livestock each year in Germany alone. Likewise, some 300 metric tons of antibiotics are used to treat humans each year, far too often for those merely suffering from a common cold. Only four pharmaceutical companies worldwide are still working on developing new agents because of the high costs of testing a drug in human subjects.


Antibiotic Action, a global alliance for antibiotic drug discovery [267]

At the World Health Day, 2011, antimicrobial resistance and the demise of antibacterial drug discovery was discussed focusing on Antibiotic Action, a global alliance for antibiotic drug discovery and development. Antibiotic Action is based on academic-industrial partnerships, licensing and regulation of new antibiotics and economics of antimicrobial drugs. [268]

Genetic risk of type 2 diabetes mellitus

[269] Diabetes is widespread in industrial countries. It develops mainly in advanced age, but children may also have type 2 diabetes mellitus. Overweight, lack of physical exercise and genetic factors may trigger the disease.

It is supposed that the body on behalf of continuously high blood-sugar, produces so much insulin that the cells do not react any more on this hormone and at some moment the produced insuline is not sufficient any more. Type I diabetes is somewhat of an autoimmune reaction working against the production of insulin of the pancreas.

Robert Sladek and colleagues from the McGill University in Montreal identified four loci in the human genome, containing variants that confer type 2 diabetes risk, in addition to confirming the known association with the TCF7L2 gene.

These loci include a non-synonymous polymorphism in the zinc transporter SLC30A8, which is expressed exclusively in insulin-producing beta-cells, and two linkage disequilibrium blocks that contain genes potentially involved in beta-cell development or function (IDE-KIF11-HHEX and EXT2-ALX4). These associations explain a substantial portion of disease risk and constitute proof of principle for the genome-wide approach to the elucidation of complex genetic traits.

The researchers are developing an easy test which can identify the genetic anomaly and direct the therapy toward the modified zinc transporter. The authors look forward to a genetic test of persons with the predisposition to type 2 diabetes. This can motivate these persons to a follow a specific nutrition plan and practice more sports and avoid the development of the disease.

Genetic factors of obesity

Gene regulating obesity, type 2 diabetes and cardiovascular diseases [270]

Researchers identified the MCPIP gene to regulat the fat cell formation and blood vessel formation that feeds the growing fat tissue. Leading author Pappachan Kolattukudy stresses that targeting this gene, a drug may shut down its function, preventing obesity and inflammatory diseases resulting from obesity, including diabetes and cardiovascular diseases.

The peroxisome proliferator-activated receptor gamma (PPAR gamma), had been known to control fat cell formation which could not be induced by other factor without the PPAR gama gene. Kolattukudy and colleagues silenced the PPAR gama gene and introduced the MCPIP gene to living cells from mice. The cells were still capable of fat formation. The increased inflammation of fat cells is seen to reduces their sensitiveness to insulin, increasing the risk of type 2 diabetes. The authors underline that MCPIP can induce adipogenesis without PPARgama.

Genes acting in the central nervous system are linked to predisposition to obesity [271]

The two loci, FTO and MC4R are two loci which are associated with body mass index (BMI) in humans. Hirschhorn and colleagues 2008 identified six additional loci: TMEM18, KCTD15, GNPDA2, SH2B1, MTCH2 and NEGR1. Several of these genes are known to act in the central nervous system (CNS) which is linked to predisposition to obesity, demnstrating the neuronal influence on body weight regulation.

FTO gene linked to a predisposition of persons to become obese [272]

Rüther and colleagues 2008 report that FTO gene is linked to a predisposition of persons to become skinny or obese. The authors stress that leptin, and adiponectin, were less expressed in mice with Fto, compared with mice whose Fto gene were silenced. These two hormones control appetite, energy balance and metabolic processes. Lower levels of these hormones are associated with obesity.

The leanness of Fto-deficient mice was found in this study to result of an increased of energy expenditure and systemic sympathetic activation, despite decreased physical activity and high food intake. The authors suggest that the Fto gene is energy regulation whith increased energy expenditure.

Genome-wide association scans to study complex diseases, such as obesity [273]

Hebebrand, Bammann and Hinney 2010 suggest the introduction of genome-wide association scans as a tool for the study of complex diseases. The receptor variant with the amino acid isoleucin (wildtype: valine) at position 103 of the melanocortin-4 receptor (MC4R) also the polymorphisms located 188 kb at the 3' end of the MC4R are cited by the authors as examples of such polygenic variants with an effect on body weight. A strong polygenic obesity effect was also coupled with variants in the first intron of the "fat mass and obesity associated" gene, also 17 polygenes for body weight regulation were described in December 2009.

The identification of the satiety hormone leptin was the first molecular genetic finding used in obesity treatment. Further studies found that a polygenic variant by itself has a small effect and only the combination of various factors lead to obesity.

Variants in the first intron of the "fat mass and obesity associated" gene result in an elevated body mass index (BMI) were detected in a genome wide association study pertaining to type 2 diabetes mellitus [274].

Polygenic obesity [275]

According to Hinney and Hebebrand 2008 only few genes are coupled with obesity. Mutations are infrequent and are therefore of minor importance. Genetic predisposition to obesity is not linked to a single gene, but is polygenic. Such genes are also found in normal weight persons, a polygene must therefore be determined by statistical analyses, such as found of Val103 allele which is more frequent in obese individuals. The INSIG2 polymorphism in body weight regulation, SNP alleles in intron 1 of the fat mass and obesity associated gene (FTO) are also being suggested to exert polygenic effect on obesity and variation in FTO strongly contributes to early onset obesity.

Clement, Boutin and Froguel 2002 stress that obesity is a multifactorial disease with interaction of environmental and genetic factors. Only rare cases of severe obesity determined by a single gene are known. Exceptional mutations of the leptin gene and mutations in the melanocortin receptor 4 were described. Obesity genes encode proteins connected with the regulation of food intake. The most common forms of obesity are polygenic. Genome-wide scans identify chromosomal regions showing linkage with obesity [276].

Findings of the Metabolism Initiative and Program in Medical and Population Genetics [277]

Speliotes and colleagues 2010 confirmed 14 known obesity susceptibility loci and identified 18 new loci associated with obesity which are located near key hypothalamic regulators of energy balance, and near GIPR, an incretin receptor.

Two new obesity loci TNKS and MSRA [278]

Scherag at al 2010 analysing two Genome-wide associations (GWAS) of extremely obese children and adolescents found two new obesity loci which join the previously identified FTO, MC4R, and TMEM18 loci. One was SDCCAG8 (serologically defined colon cancer antigen 8 gene); and one between TNKS (tankyrase, TRF1-interacting ankyrin-related ADP-ribose polymerase gene) and the second new loci was MSRA (methionine sulfoxide reductase A gene) were found by the authors. All these loci are associated with early-onset obesity.

Viral etiology of obesity [279]

Dhurandha and colleagues 2009 report that the human adenovirus Ad36, may promotes obesity. The study used rodent preadipocyte cell line. A nonadipogenic human adenovirus Ad2 was used as a negative control for viral infection. Ad36, but not Ad2, was found to accumulate lipid in cells.

Ad36 downregulated preadipocyte marker gene Wnt10b, and upregulated expression of early (C/EBPDelta and C/EBPbeta), intermediate (PPARgamma2), and late genes (aP2 and G3PDH) of adipogenic cascade. The authors concluded that Ad36 is an exogenous regulator of the adipogenic process.

The researchers van Ginneken, Sitnyakowsky and Jeffery 2009, following the results of the studies of Dhurandha and colleagues, postulate that infection with Ad-36 may be another factor of the global rising obesity. According to the authors, viral infections may add another cause of obesity to the long recognized factors such as genetic inheritance, diet, exercise, cultural practices and stress. Four viruses have been reported to induce obesity in animal, however these viruses were unable to infect humans. The human adenovirus Ad36 is now recognized to affects human primary preadipocytes, and antibodies to Ad-36 were more prevalent in obese subjects (30%) than in non-obese subjects (11%). [280]

Enzyme controlling obesity [281]

Sul and colleagues 2009 report that an enzyme, the adipocyte phospholipase A(2), AdPLA (encoded by Pla2g16, also called HREV107) regulates lipolysis and adiposity. The authors found that mice without AdPLA have a markedly higher rate of lipolysis have markedly reduced adipose tissue mass and triglyceride content, and higher energy expenditure with increased fatty acid oxidation. They eat more while remaining lean.

The researchers concluded that AdPLA enzyme is a major regulator of fat physiology and is an important factor of obesity.

Antibiotic resistance marker genes nptII in GM plants poses no risk to human, animal or environment

[282] The nptII gene (neomycin phosphototransferase II) provides resistance to certain antibiotics, such as the aminoglycosides kanamycin, neomycin and geneticin, in plants. In in 2004 the EFSA found no scientific evidence that a transfer of this gene to bacteria, animals or humans could occur.

The EC asked EFSA to consider an information of EMEA and any potential consequences for the safety of the nptII gene as a marker gene in genetically modified plants and for previous opinions on specific GM plants (and derived food and feed) comprising the nptII gene.

According to the European Medicines Agency EMEA information, aminoglycosides comprise a class of antibiotics that has become increasingly important in the prevention and treatment of serious invasive bacterial infections in humans. This is because bacteria, such as tuberculosis bacteria, are becoming resistant to other classes of antibiotics. It also said that, although kanamycin and neomycin are used relatively infrequently, the development of similar antibiotics should be taken into account and that aminoglycosides as a group are a class of antibiotics critically important for veterinary medicine.

The GMO Panel responded agreeing with the EMEA that the preservation of the therapeutic potential of the aminoglycoside group of antibiotics is important. The Panel is also of the opinion that the therapeutic effect of these antibiotics will not be compromised by the presence of the nptII gene in GM plants, given the extremely low probability of gene transfer from plants to bacteria.

The GMO Panel reconfirms its earlier conclusions in its opinion on antibiotic resistant marker genes that the use of the nptII gene as selectable marker in GM plants (and derived food or feed) does not pose a risk to human or animal health or to the environment.

The FSA arguements

Likelihood of transfer of the nptII gene from the genome of GM plants to bacteria [283]

(i) DNA is released from plant material by normal digestion processes that take place in the gastrointestinal tract, or by activities of nucleases present in various organisms in the environment.

(ii)The probability that bacteria will be exposed to DNA stretches long enough to contain the intact nptII gene is very low because of the above mentioned digestion and degradation processes.

(iii) The nptII gene from plant material can only be taken up by competent bacteria via natural transformation, a process that occurs infrequently in many bacteria and in most environmental conditions.

(iv) If the intact nptII gene enters the bacteria, it will be rapidly degraded by restriction endonucleases inm many bacterial cells which possess DNA restriction systems in order to destroy foreign DNA.

(v) If the intact nptII gene does indeed survive, the probability of its incorporation into the bacterial genome is very low unless there are homologous regions already present in the bacterial genome. Gene transfer from plants to bacteria has only been demonstrated under laboratory conditions when regions of homology were already present in the recipient bacterium.

(vi) Expression of the incorporated nptII gene is unlikely considering that in GM plant material the nptII gene is under the control of a promoter with preferential expression in plants, which does not support its efficient expression in bacteria.

(vii) Stable integration and inheritance of the nptII gene in the host bacterium is not likely in the absence of selective pressure from a relevant antibiotic.

When all of the above mentioned aspects are taken into account, the probability of functional gene transfer from plants into microorganisms is extremely low. It is not surprising that transfer of an antibiotic resistance marker from GM plants to bacteria has not been observed under natural conditions.

The EMEA has indicated that under laboratory conditions gene transfer from plants to bacteria has been demonstrated. According to EFSA, however, gene transfer from plants to bacteria has only been demonstrated in a few highly transformable bacterial species (e.g., Acinetobacter sp. BD413 or Pseudomonas stutzeri) under artificial and forced laboratory conditions when regions of homology were already present in the recipient bacterium. In the absence of this optimisation of the process and selection pressure, resistance gene transfer from GM plants to bacteria, even in the laboratory, could not be demonstrated.

Prevalence of the nptII gene in soil, humans and animals [283]

Antibiotic resistance is a common feature in natural microbial communities in soils, aquatic systems, and habitats associated with animals and humans. There is already a widespread presence of nptII in the soil environment.
Studies indicate that, as expected of a gene located on a transposable genetic element, nptII is located on a wide range of replicons in bacterial clinical isolates from humans The nptII gene was present in 2.5% of bacterial clinical isolates resistant to kanamycin and neomycin collected between 1987 and 1991 in several European and Central and South American countries (Shaw et al., 1993). Studies on the prevalence of the nptII gene in animal-associated bacterial populations have not been found in the scientific literature.

Contribution of the nptII gene to the prevalence of resistance to kanamycin [283]

Kanamycin-resistant bacteria are ubiquitous in nature. Only a fraction of kanamycin-resistant bacteria contain the nptII (aph(3')-IIa) gene, the other resistant bacteria having different genes and/or other mechanisms conferring kanamycin resistance.

The nptII gene has been reported to occur naturally only in eubacteria. In one survey, 3 out of 184 kanamycin resistant bacterial isolates from three stream sites in the USA (Leff et al., 1993) and 44 out of 355 from different habitats in the Netherlands (Smalla et al., 1993) contained nptII sequences.

Potential mutations of the nptII gene resulting in resistance to other antibiotics [283]

Resistance towards amikacin, an important reserve antibiotic could be obtained under laboratory conditions and was the result of a mutated nptII gene and a diminished rate of amikacin uptake into the bacterial cell. However, to date no clinical amikacin resistant strains with a mutated nptII gene have been identified.

Rice genetically modified with high flavonoid content [284]

Researchers from the from Hamburg University and the University of Hyderabad (India) found in 2007 GM rice with high antiooxidant activity. The rice contains the anthocyanidin synthase (ANS) enzyme. ANS enzyme is involved in the biosynthesis of flavonoids, transforming leucoanthocyanidins into coloured anthiocyanidins. The transgenic rice 10TC is a mutant strain of the rice called Nootripathh.

The authors compared the favonoid content of the transgenic Rice with that of normal non GM rice:

Flavonoids Produced in 10TC Rice Normal nonGM Rice
  micrograms per milligram micrograms per milligram
     
Anthocyanins 2.52 0.12
     
     
Quercetin 1.37 0.55
     
     
Proanthocyanidins 0.09 0.40
     



The content of flavonoids had 22% higher antioxidant activity than untransformed rice. The authors claim that their method can be used to enhance the nutritional value and resistance against biotic and abiotic stresses of different food crops. This would strongly support genetic modification of staple foods.

Defra final decision on BASF GM potato trial postponed [285] [286]

The UK's department for the environment, food and rural affairs (Defra), which originally gave approval in December for BASF to undertake research trials of a GM potato at two sites in England, one in Cambridgeshire and the other in Derbyshire, said that it would now consider BASF's proposal as a new application in accordance with the Genetically Modified Organisms (Deliberate Release) Regulations 2002. Similar trials are already underway in in Sweden, Germany and the Netherlands. But before reaching a definitive decision, Defra said that it would consider any representations that people may wish to make about the risk of environmental damage posed by the GM trial. The deadline for representations is 20 April 2007.

Defra announced in its News Release from 01.12.06 the approval of an application of BASF to undertake trials of a late potato blight disease GM potato on two sites in England, starting in 2007.Evaluation of the application by the Advisory Committee of Releases to the Environment (ACRE ). found that the trials will not result in any adverse effect on human health or the environment. [287]

Clare Oxborrow Friends of the Earth says that the trials pose a significant contamination threat to future potato crops, claiming that there is no need for GM potatoes and no consumer demand for them. This was backed by Lord Peter Melchett from the Soil Association warning that other crops risk contamination by GM. [288]

Professor Philip Dale of the plant-breeding John Innes Centre argued that the Soil Association is opposing this project because the Association see these kinds of advances in general agriculture to be a threat to the profitability of organic farming.

GM rice with human genes

In 2004 the company, Ventria Bioscience, Californis, started to cultivate rice engineered to produce lactiva and lysomin. These proteins are found in breast milk and should improve recovery from diarrhoea. [289] The company has now received approval from the US Department of Agriculture to cultivate this rice in the state of Kansas. According to Robert Wittler, from Kansas University School of Medicine at Wichita the outcomes of a clinical trial using the new rice suggests that children suffering from diarrhoea may recover sooner compared with traditional traditional medication. [290] GeneWatch UK and Friends of the Earth, point out the danger of a broad contamination of staple foods with pharmaceutical producing genes. The genes, cultivated and copied in a laboratory to produce a synthetic version, are carried into embryonic rice plants inside bacteria. [291]

Pharmaceutical rice

[292] [293] [294] The California Rice Commission was created by the government of California to serve the interests of the California rice industry by expanding and maintaining the industry's markets. California ranks second in the United States behind Arkansas in rice production, accounting for 20% of the U.S. total production.

In 2000, the California legislature, in response to a proposal advanced by the CRC, enacted the Rice Certification Act of 2000279 with the broad intent of enhancing and protecting the reputation of California's rice industry throughout the nation and around the world.

Ventria [295]

Lonnerdal in a review states that expression of recombinant human milk proteins in rice is realistic and a possibility for the addition of bioactive factors to infant formula and baby foods. The protein composition of infant formula is still quite different than that of human milk, as there are proteins in cow's milk which are absent in human milk, and many proteins in human milk are absent in cow's milk such as lactoferrin. Addition of human milk proteins to infant formula may be necessary to obtain some of the nutritional and health benefits that breast-fed infants enjoy.

Recombinant human milk proteins can now be expressed in a variety of systems.

Recombinant human lactoferrin and lysozyme added to a rice-based oral rehydration solution were found to increase recovery of children with acute diarrhea. [296]

Ventria rice expresses human lactoferrin or lysozyme protecting the intestinal tract similarly to subtherapeutic antibiotics. The GM rice has potentials to to be used as a substitute for antibiotics in broiler diets. [297]

Saccharomyces is used for expression of human lactoferrin but has low expression levels. Aspergillus is being used in the production of recombinant human lactoferrin but the cost is too high for use as a food additive. [295]

Ventria and its GM rice [292]

In the fall of 2002, Ventria began formal discussions with the CRC on its intent to commercially plant its pharma rice during the 2004 planting season, and, in December 2003, Ventria submitted an application to APHIS to renew its California field trial permits.

In order for Ventria's pharma rice product to be commercialized in California, APHIS would have to authorize the necessary planting through the issuance of an appropriate permit. Of the 84 permits for the field testing of pharma crops that APHIS has issued nationwide, nine have been issued for trials in California, including for pharma rice, but APHIS has not to date authorized commercial production for any pharma crop.

Ventria stipulated that its pharma rice had characteristics of commercial impact. The advisory board of CRC worked with Ventria to develop proposed conditions for the production and handling of the company's pharma rice. These included growing the pharma rice in Southern California, which is outside the state's rice belt; not seeding the rice from the air, ensuring a buffer zone of 100 feet between biotech rice and other crops; ensuring seed containers are sealed and numbered and silos are labeled and locked in order to keep pharma rice separate from other rice; and testing for the presence of the biotech pharma trait.

On March 29, 2004, by a vote of six to five, the advisory board recommended to the secretary of agriculture conditions and protocols under which Ventria's pharma rice could be planted commercially with adequate identity preservation and containment.

California Department of Food and Agriculture CDFA Secretary Kawamura denied the recommendation of the commission's advisory board for an emergency exemption to plant its pharma rice crop during the spring 2004.

Ventria said the company has plans to reapply in California and is also considering other options, such as planting in Hawaii and states in the South.

Japans concern about the US pharma rice

Japan imports 40% of the California's production of rice. The Japanese Rice Retailers Association has been concerned about biotech products in general and the commercialization of pharma rice specifically which will evoke a distrust of U.S. rice as a whole among Japanese consumers,

The argument supporting pharmaceutical rice

The Biotechnology Industry Organization (BIO) argues that the health benefits of pharma crops outweigh the risks and that the risks are adequately regulated by the federal government.

The industry also points out that food crops are good platforms for pharma crop development and production.

Acording to Ventia the health benefits of the technology outweigh the risks, claiming that producing these proteins through crops is the most cost-effective and efficient means of reaching the most people. Planting 65 acres of pharma rice, they say, could generate 1,400 pounds of lactoferrin, which would be enough to treat 650,000 children with dehydration, a condition that kills 3 million infants each year worldwide, mostly in developing countries.

However, a report by the National Research Council on the biological containment of genetically modified organisms concluded that crops used to produce common food products would be a "poor choice" for use to produce pharma and industrial crops unless they can be grown under "stringent conditions of confinement."

The USDA's Animal and Plant Health Inspection Service (APHIS) had discovered GM in a variety of long grain rice seed from BASF, all of them outside of the California state. Following this announcement, the California Rice Commission (CRC) called for a suspension of field testing of all genetically modified rice cultivars in the State of California for the 2007 crop, and for future crops, until safety is guaranteed.

These arguments made the California Rice Commission, to call for a moratorium on GM field testing in the state, allowing for an opportunity to evaluate federal regulations that safeguard the rice industry.

Reduction of Pressure on Palmoil production [298]

John Shanklin looking for better understanding of the enzymes and metabolic pathways that produce oils in oil plants found how to manipulate the accumulation of fats using genetic techniques. They found that reducing the levels of the gene Arabidopsis KASII convert temperate oilseed composition of temperate oil plants , such as canola, soybean, and sunflower to that of a palm-like tropical oil.

Temperate oil add two carbons with a desaturating activity of the expression of the gene KASII. Reducing the expression of this gene the double bounded elongation of the fatty acid chain from C16 to C18 is reduced. A higher melting point of the fat is attained which resembles that of palm oil.

This may reduce the pressure on the forests in Asian countries saving thus precious wildlife.

New oil palm hybrids to reduce pressure on Indonesian forests [299]

Environmental concerns about the destruction of Indonesian forests and peatlands, increasing greenhouse gas emissions and damaging habitats for animals are driving big corporations to develop new varieties of oil palm plants which present higher yields and are more resistant to diseases. The University of Reading, working with Sumatra Bioscience, BioHybrids International Limited, and the University of Aberystwyth are developing F1 oil palm hybrids which are are more economically viable and environmentally sustainable. The F1 hybrid from Sumatra Bioscience is to be marketed by 2018. The F1 oil palm hybrids are the result of crossing two distinctly different oil palm parental lines. These lines are genetically uniform, resulting in higher yield per unit area and may be developed to present specific oil profiles to meet specific demands . Other companies, also looking for oil palm plants with higher yields are the US-based Synthetic Genomics and the Asiatic Centre for Genome Technology.

Transgenes in Mexican maize at the preliminary stage of climate change

[300] The actual discussion on the introduction of drought resistant transgenes facing the climate change may include the outcomes of the findings of Ortiz-Garcia in 2004 concerning maize transgenes in the Mexican state of Oaxaca which find no danger to maize diversity in that region.

The presence of transgenes in maize landraces in the state of Oaxaca, Mexico had been reported by Quist and Chapela [301] in 2000, but there were doubts the methodology used. However, he Mexican government confirmed the presence of transgenes in Oaxaca in 2000 and 2001 [302] [303]. Ortiz-Garcia et al.in 2003 and 2004 [304],found no transgenes in the area. It is presumed that their frequency had diminished greatly over the course of 2-3 years, and the genes may even have disappeared.

Peter Raven believes that whether or not transgenes are present in landraces in Oaxaca at present, they will inevitably be found in them as time passes, because of the nature of the indigenous agriculture. There they will persist if they confer a selective advantage on the plants in which they occur, or they may disappear if they do not confer such an advantage in the prevailing conditions.

As Ortiz-Garcia et al. [304] have pointed out, it is unlikely that the presence of transgenes could reduce the genetic diversity of the landraces in which they might occur. In general, for the landraces of maize in Mexico or for any other populations, their genetic characteristics should remain essentially unchanged unless there is strong selection for whole constellations of characteristics from radically different strains of maize, conditions that have not been observed in southern Mexico.

Raven concludes that the introduction of the transgenes currently in use for maize poses no danger to maize near its center of origin, to the Mexicans, or generally.

Soy gene GmNAC6 of the endoplasmic reticulum (ER) responding to drought and salinity [305]

Faria et al 2011 describe a coordinated and integrated molecular network for stress signal to adapt to adverse conditions. Responses to a specific stress condition are interconnected with other environmental responses, using pathways among the organelles to respond to environmental signals.

The endoplasmic reticulum (ER) supportes basic cellular processes, such as protein folding and quality control, the maintenance of Ca2+ balance and lipid biosynthesis. Genes in this integrated pathway encode proteins with diverse roles, such as plant-specific development and cell death proteins. The N-rich protein (NRP) genes, NRP-A and NRPB induce a senescence-like response to stressors in soybean cells and tobacco leaves.

The NRP-mediated cell death signaling pathway is a ER-stress response that communicates with other environmental stressors, such as the osmotic stress signals. The authors describe a member of the NAC domain-containing protein family from soybean, GmNAC6 (Glycine max NAC6), that may act downstream of NRP-A or NRP-B in the integration of the ER-stress and osmotic-stress cell death signals. Both expressions of NRP-A and NRP-B are necessary to increase the expression of GmNAC6.

New-Zealand GM cow produces skimmed milk [306]

In an article by Cath O'Driscoll 2007 in Chemistry and Industry genetic modification of cows to produce skim milk is reported. The authors claim that special genes could be used to breed herds of animals producing only skimmed milk.

The authors say that a commercial herd will be available by 2011. The research is being done by the biotech firm ViaLactia at the dairy co-operative Fonterra which already makes milk from one of the GM cows which was named Merge. The new GM cow will fit very well into the good selling market of low fat products.

According to the authors this could reduce the surplus of cream and there would be no need for disposal. However, the cream is used to make butter and other products such as ice cream, confectioneries,and bakeries.

Another exciting development of their cows they are working on is to produce a GM cow which gives butter that is spreadable straight from the fridge. Experiments to soften butter using special feed are not so successful as the change to GM cows.

Detection of L and D amino acids to study modified composition of new GM plants [307]

Miguel Herrero and colleagues 2007 presented a new technique detecting significant differences% D-aa values for the different conventional varieties (Aristis, Tietar, and PR33P66 maize).

The transgenic maize Tietar-Bt with the new inserted Cry1Ab transgene presented % D-aa values which were similar to conventional maize Trietar. According to the authors this indicates that, in these maize samples, no metabolic pathway were modified in Trietar-BT.

The method is based on changes of the "L" or "D" form of amino acid (the so called chiral amino acids). Such changes may affect nutritional quality and digestibility. The method combines micellar electrokinetic chromatography with a chiral selector and laser-induced fluorescence to investigate the prevalence of L- and D-amino acids.

The authors suggest this new method to be used as proof of substantial equivalence of GM maize and conventional plants.

EU rises the threshold of "GM Free" labelling [308]

Natural normal grown food is acute endangered to be contaminated by the genetic engineering escapades of a handful global enterprises. Governments bow their head to their lobby and rise the threshold labelling of contamination instead of combating the underlying cause which would be to phase out GM food.

EU agriculture ministers last week agreed that food accidentally contaminated with up to 0.9% genetically modified content could be labelled "GM free". But the Soil Association and Organic Farmers and Growers, which together certify more than 90% of the UK's organic food, today pledged to keep their own criteria at the current 0.1 per cent.

On 21 June 2007, a delegation representing 70 organic businesses, urged the Secretary of State for the Environment not to make organic consumers pay for GM contamination. The Government currently proposes to allow up to 0.9% GM in organic food without the GM companies being liable for this damage. This is despite the fact that in European law, 0.9% is not a general GM threshold, but rather a threshold below which accidental contamination does not have to be labelled.

The UK ognanic associations confirmed that they would maintain their current non-GM standards for organic food, at the effective minimum detectable level of 0.1% GM, whatever the Government decides.

During their consultation, the Government met with a number of GM companies, including AstraZeneca, BASF Plant Science, Bayer CropScience, Dow AgroSciences, Du Pont (UK) Ltd, Monsanto UK Ltd, and Syngenta Ltd. Not one organic business was consulted directly, and this is the first meeting the Government has had with representatives of organic businesses.

EU Council of Ministers postpones its decision on approval of GM Amflora potato from BASF:

On the 29. July 2007 the Council of Ministers postponed the decision over whether GM potato Amflora may be commercially cultivated in Europe in December.

On 14. July 2007 the trade commissioner Peter Mandelson delivered a strong exhortation to the EU to take a lead in shaping global rules on GM trade warned about the economic consequences. Na statesman can take the risks of acting hastily and place financial and trade interests in front of safety. The trade commissioner Mandelson would be well advised to look after his trade and leave the biotechnology to the board of scientific advisers whose job is to look after food and environmental safety.

Amflora is deeply modified in the starch it produces. Non-GM potatos produce two components starch. 80% are amylopectine with a long molecule with thickening properties. and 20% amylose with short molecule with gelling character. Amflora produces only amilopectin, which is more useful for the industry as amylose.

Amflora, the GM potato for adhesives and paper industries [309]

The first EU authorised harvest of the BASF potato "Amflora" cause a new discussion concerning the need of genetic modification in modern agrarian technology.

BASF's Chairman Dr. Jürgen Hambrecht announced that BASF has also applied for approval of its second starch potato named Amadea. The high-performing starch potato which produces pure amylopectin starch is expected to be launched in 2013/14.

Amflora and Amadea are two genetically modified potato varieties that produce pure amylopectin starch, being superior to conventional potatoes which produce a mixture of amylopectin and amylose, but only amylopectin is needed in the production of adhesives or paper. Separating amylose from the starch of conventional potatoes is an additional costly process.

Amylopectine improves adhesives. As a coating of paper mylopectine improves the paper gloss. Karl-Heinz Niehoff is a German farmer which plants potatoes for the food industry for French-fries, but also starch enriced potatoes for the starch industry. Nienhoff platen 15 hectares of Amflora contracted by BASF Plant Science. One hectare Amflora crop was destroyed at night by environmental activists.

Nienhoff says that the GM potato of BASF might improve its marketability in competition with corn starch. The price of potatoes is guaranteed by the EU until 2012. Alternatives are needed, because without this support the cultivating potatoes will be more difficult.

Safety of GM potato

The GM potato is less prone to migrate between non-GM fields because it reproduces itself only by tubers and not by seeds or by pollen as seen with rape and by corn.

However, a mix-up of Amadea and Amflora occurred at the facilities of BASF and a potato crop of 15 hectares had to be destroyed in Sweden in August 2010, demonstrating that migration of GM plants to nature is unavoidable. [310]

Three years of Amflora trial plantations seem to go to an end. Conventional breeders succeeded, meanwhile, to create an amylopectin-rich potato using conventional crossing methods which will take the place of the GM potato. The farmer Nienhoff says he will not plant the GM potato any more.

German conventional potato receives Geographical Indications protection by the EU [311]

Potato-growing has a long tradition in the Lüneburger Heide. Since the second half of the 19th century the Lüneburger Heide has been one of the best-known and largest potato-growing areas in Germany.

The European Commission has added "Lüneburger Heidekartoffeln" to the Register of Protected Designations of Origin (PDO) and Protected Geographical Indications (PGI):
The EU Commission "Lüneburger Heidekartoffeln" have acquired considerable renown and a good reputation in the region and beyond. The varieties grown in the Lüneburger Heide ((Germany) are known as "mainly firm boiling" and "firm boiling" potatoes, and are healthy, whole, firm and practically clean.

BASF increases GM expansion in Asia [312]

According to BASF the corporation signed cooperation and licensing agreement in biotechnology with Korea and China on GM crops such as corn, soybeans and rice. It will use a family of genes, such as license rights from CropDesign, which increase crop yield and can be transferred into a range of other crops.

Asia Pacific due to its emerging economy avoids everything which curbs quick development regardless to environment, ecology and food safety. One of these rapid development experiments had been the Green Revolution in India which is now driving local peasants to suicide. It is wellknown that food politics in China is very far from excellent and China food is being warned from, so the GM traits of BASF suits well in this market.

GM maize Herculex RW import approval on the EU agenda

Representatives from the 27 EU member states will vote on the 25 of June 2007 to authorise imports GM Maize 59122 known as Herculex RW with the bt protein resisting the maize rootworm. Approval will be for use in food, feed and processing.

Herculex RW has been planted for the first time in US in 2006 and was approved. Australia, New Zealand, Canada, China, Japan, Korea, Mexico, Philippines and Taiwan.EFSA approved it in March 2007. Only Austria and Luxenburg may block the vote. Overrunning Austria and Luxenburg will not be to difficult for Herculex RW which will contaminate the whole European feed (meat, eggs, aquaculture), the glucose-fructose syrup (broadly used in all fruit yoghurt and dairy products, bakery products).

The European Green Party is against the authorisation claiming that there are serious and legitimate health concerns about Herculex RW maize as changes in the blood parameters and in liver weight in rats fed with this GM variety were found in a research.

This is being sustained by the Italian Green Party and the Italian farmers' union which also protested against the plans to test GM versions of GM olives and tomatoes. Organic associations strongly protest against an increasing contamination of their products by GM intrusion.

New germplasm lines for breeding sunflowers [313]

To improve plant resistance of sunflower plants to downy mildew, caused by the fungus Plasmopara halstedii. the USDA's AgriculturalResearch Service (ARS) and North Dakota Agricultural Experimen developed the HA 458, HA 459 and HA 460 lines crossing elite sunflower lines with wild plants collected from Idaho and Texas.

The downy mildew fungus affects seedlings and mature plants. The oleic fatty acid has been increased, resulting an increased frying stability.

In 2003 the Archer Daniel Midland's NuSun, a mid-oleic oil became commercially available. It has less than 10 per cent saturated fat and oleic levels of between 55-75 per cent.

According to ARS, 15 new races of downy mildew developed resistance to the fungicide metalaxyl, the new germplasms will acquire outstanding imortance to reduce crop losses.

In 2004 it released three others, known as RHA 439, RH 440 and HA 441, intended to fight sclerontinia sclerotiorum, a disease that causes stalk and head rot in the plants.

The oleic acid concentration of HA 444, HA 445, and HA 446 was 83.6, 85.9, and 81.7%, respectively, compared with the oleic acid concentration of 86.1% for HA 434. [314]

ARS plant pathologist Tom Gulya is researching the integrated control of Sclerotinia wilt and headrot, and control of metalaxyl-resistant downy mildew through new resistance genes complimented with the new fungicides. Other research topics include finding resistance to a new strain of Verticillium wilt and continued work on Phomopsis stem canker. [315]

Genetic modified carrots could enhanced calcium intake [316]

Jay Morris and colleagues 2008 modified genetically carrots increasing the absorption of calcium by 41 per cent in humans.

According to the authors the new calcium enriched carrot could help to compensate for a worldwide nutritional calcium deficiency.

The increased absorption was attained by altering the sCAXI gene which controls the absorption and transport of calcium in plants.

The authors stress that the technology may be applied to various crops because it involves the over-expression of a gene found in all plants.

GM kiwi fruit will meet flavour, colour and market demands, says New Zwaland research [317]

The New Zealand's Hort Research and Genesis Research and Development Corporation in 2008 published a collection of 130,000 DNA sequences of kiwi fruit, which enables breeders to tailor new fruits to meet consumer tastes and market needs. The expressed sequence tags (EST), from plant genes expressing flavour, colour, shape, and vitamin content, ripening and storage time are now identified.

The authors stress that using Marker Assisted Selection (MAS) breeders may identify the specific genes of interest. The outcomes of this research will be used in the New Zealand kiwi production which amount up to 34.000 tonnes per year.

Research reveal how cells determine the protein production [318]

Silent mutations

Silent mutations are DNA mutations that do not result in a change to the amino acid sequence of a protein. They may occur in a non-coding region (outside of a gene or within an intron), or they may occur within an exon in a manner that does not alter the final amino acid sequence.

Synonymous mutation

The phrase silent mutation is often used interchangeably with the phrase synonymous mutation; however, synonymous mutations are a subcategory of the former, occurring only within exons [319].

Plotkin et al., 2009, found a hidden code that determines the expression level of a gene, providing a way to distinguish efficient genes from inefficient ones. The research revealed how a cell each protein produces the amount of proteins needed to maintain homeostasis. The researchers concluded from their data that synonymous mutations determine mRNA folding and thereby the eventual protein level.

According to the authors synonymous mutations do not alter the encoded protein, but they can influence gene expression. The researchers created 154 of synthetic green-glowing genes. all encoding the same green fluorescent protein (GFP). The mRNA folding was found to determine the amount of proteins produced by individual genes, whereas codon bias influences global translation efficiency and cellular fitness. They explained that the inefficient genes produced tightly folded mRNA molecules that could not be accessed by the protein-making machinery.

Environmental conditions induce changes in pathogenesis of Salmonella [320]

Researchers found that the expression of genes required for invasion is tightly regulated by environmental conditions and a variety of regulatory genes in Salmonella enterica serovar Typhimurium. The hilA regulator activates the expression of invasion genes in response to both environmental and genetic regulatory factors. The regulation of hilA expression is a key point for controlling expression of the invasive phenotype.

According to Jones 2005 positive regulators of hilA expression were found to be csrAB, sirA/barA, pstS, hilC/sirC/sprA, fis, and hilD and HilD and negative regulators are hilE, hha, pag, and lon.

The author says that the HilE gene Salmonella-specific regulator inactivates the hilA expression. The work of the author focuses on how the environmental signals that affect hilA expression of invasiveness may act through the hilE pathway.

Spaceshuttle missions improve unterstanding of virulence of Salmonella [321]

The spaceflight environment induces novel changes in microbial pathogenesis not observed using conventional methods. Nickerson et al. report that spaceflight-induced changes increase the virulence of Salmonella regulated by media ion composition, and that phosphate ion is sufficient to alter related pathogenesis responses in spaceflight conditions.

The authors identified evolutionarily conserved molecular pathways in Salmonella that respond to spaceflight. These findings open new ways to control microbial responses during the infection and improve vaccines.

Short DNA segments called pyknons are found to be highly important [322]

Cooper and colleagues 2008 found that intergenic DNA playing a physical role in protecting and linking chromosomes are accompanied by short segments of DNA which seem to be coding DNA. These non-random sequence patterns are called pyknons. They have the same sequence and size as small segments of RNA that regulate gene expression through gene silencing.

Pyknons had been discovered only in human genome. Analysing the genome of the plant Arabidopsis thaliana the author found pyknons which are similar to the human pyknons. The authors stress that Arabidopsis thaliana has approximately the same number of genes as the human genome but a higher number of their genes are silenced. The authors suggest that pyknons are involved. A better understanding of the effects of pyknons may be applied in the improvement of transgenic plants by regulating gene silencing.

Pyknons play a role in the evolution of human genome [323]

Non-coding DNA outside of genes had been considered material without biological function, such as tandem repeats, short stretches of DNA and are now called pyknons.

The repeats determine how tightly the local DNA is wrapped around specific proteins called 'nucleosomes', and this packaging structure dictates to what extent genes can be activated. These tandem repeats are very unstable and the changes affect the local DNA packaging, which in turn alters gene activity. This allows fast shifts in gene activity to match changing environments boosting the evolution.

Experimenting with yeasts cells of Saccharomyces cerevisiae, the authors found that when a repeat is present near a gene, it is possible to select yeast mutants that show vastly increased activity of this gene. However, when the repeat region was removed, this fast evolution was impossible. only cells with the repeats would be able to swiftly adapt to changes, thereby beating their repeat-less counterparts in the game of evolution. As many as 25% of all gene promoters contain tandem repeat sequences.

Sequencing Method confirms importance of small DNA structures unknown before [324]

Kim, Waterman and Li 2007 infered a complete sequence of the chromosomes of Ciona intestinalis, a marine invertebrate, from existing sequencing data. The applied method focused on the high rate of genetic mutations in this organism. Other organisms with high genetic variability, such as certain fish, also may be suitable. The method is not suitable to be applied to the human genome because of the low human mutation rate.

However, sequencing parts of the human genome that display high variability may be possible wqith this methode. The authors stress that small DNA structures, called pyknons, may express proteins which may regulate gene function, and were highly conserved during evolution.

Tiny RNAs Controlling Genes [325]

In 2006 Yin and Lin discovered tiny piRNAs structures in mammalian reproductive cells which may control gene functions. In a study published in 2007 both scientists found more than 13,000 Piwi-associated piRNAs in fruit flies. The most prominent piRNA was found to form a complex with the Piwi protein which binds to chromatin which controls the activity of the gene.

The authors explain that the role of chromatin is to package DNA so that it will fit into the cell, to strengthen the DNA to allow cell division, and to serve as a mechanism to control gene expression. The authors call for more researches on the silenced part which comprises 99% of the genome. These studies may be important to understand essentials for germline stem-cell maintenance.

Bone-cell control of energy generation identified [326]

Osteoblasts (bone cells) help controlling energy metabolism while expressing cell-specific regulatory genes called activating transcription factor 4 (ATF4). This gene regulates all functions linked to the maintenance of bone mass.

Karsenty and colleagues 2009 report that ATF4 inhibits insulin secretion and decreases insulin sensitivity in liver, fat, and muscle. ATF4 Atf4 increases expression of the Esp gene, which makes a protein that decreases the activity of osteocalcin. Osteocalcin is a molecule which increases the activity of insuline.

The authors concluded that endocrine functions of osteoblasts play a role in energy generation through the protein ATF4 via osteocalcin.


The Gene Ontology [327]

The Gene Ontology (GO) project is a major bioinformatics initiative with the aim of standardizing the representation of gene and gene product attributes across species and databases. The project provides a controlled vocabulary of terms for describing gene product characteristics and gene product annotation data from GO Consortium members, as well as tools to access and process this data.


Gene Ontology goals [328]

The Gene Ontology project has three major goals: 1-Develop a set of controlled, structured vocabularies-known as ontologies-to describe key domains of molecular biology, including gene product attributes and biological sequences. 2-Apply GO terms in the annotation of sequences, genes or gene products in biological databases. 3-Provide a centralized public resource allowing universal access to the ontologies, annotation data sets and software tools developed for use with GO data. http://www.geneontology.org/

Ontologies

An ontology is a description of the concepts and relationships that can exist for an agent or a community of agents. The Gene Ontology project provides ontologies to describe attributes of gene products in three non-overlapping domains of molecular biology.
Molecular Function (MF) describes activities, such as catalytic or binding activities, at the molecular level. GO molecular function terms represent activities rather than the entities (molecules or complexes) that perform the actions, and do not specify where, when or in what context the action takes place.
Biological Process (BP) describes biological goals accomplished by one or more ordered assemblies of molecular functions. High-level processes such as "cell death" can have both subtypes, such as "apoptosis", and subprocesses, such as "apoptotic chromosome condensation".
Cellular Component (CC) describes locations, at the levels of subcellular structures and macromolecular complexes. Examples of cellular components include "nuclear inner membrane", with the synonym "inner envelope", and the "ubiquitin ligase complex", with several subtypes of these complexes represented.

For example, the gene product cytochrome c can be described by the molecular function term oxidoreductase activity, the biological process terms oxidative phosphorylation and induction of cell death, and the cellular component terms mitochondrial matrix and mitochondrial inner membrane.

Several new relationship types have been introduced and used, along with existing relationships, to create links between and within the GO domains. [329]


Sequence Ontology [330]

Sequence Ontology (SO) permits the classification and standard representation of sequence features. Sequence ontology is being maintained separately from the GO ontologies. http://www.sequenceontology.org/index.html

Patents on breath Cancer genes are unconstitutional and invalid, say two civil organisations [331]

The American Civil Liberties Union (ACLU) and the Public Patent Foundation (PUBPAT) filed a lawsuit in 2009 charging that patents on two human genes associated with breast and ovarian cancer are unconstitutional and invalid.

The lawsuit was filed against the U.S. Patent and Trademark Office, as well as Myriad Genetics and the University of Utah Research Foundation, which hold the patents on the genes, BRCA1 and BRCA2. The lawsuit charges that patents on human genes violate the First Amendment and patent law because genes are "products of nature" and therefore can't be patented.

According to ACLU about 20 percent of our genes are patented. A gene patent holder has the right to prevent anyone from studying, testing or even looking at a gene. As a result, scientific research and genetic testing has been delayed, limited or even shut down due to concerns about gene patents.

The ACLU stresses that mutations along the BRCA genes are responsible for most cases of hereditary breast and ovarian cancers. The patents granted to Myriad give the company the exclusive right to perform diagnostic tests on the BRCA1 and BRCA2 genes and to prevent any researcher from even looking at the genes without first getting permission from Myriad.

Research on multi-protein mTOR opens new therapy of cancer [332]

Ruggero and colleagues 2010 studied the function of a multi-protein unit known as mammalian target of rapamycin (mTOR) which is implicated in the formation of lymphoma and other types of cancer. The mTOR protein controls the pathway of several important cell processes, such as cell survival and proliferation.

The researchers are looking at a new substance, PP242, which inhibits mTORC1-dependent 4EBP-eIF4E hyperactivation which influences the control of Mcl-1 expression, a key antiapoptotic protein. The authors are hopeful that PP242 may lead to a new therapy of cancer.

Synthetic chromosome takes over the control of bacteria [333]

Venter and colleagues report in 2010 that they exchanged the chromosome of a bacterial cell with a chromosome synthesized in their laboratory. Only available chemicals were used for the synthesis of the artificial chromosome. The bacteria with the new chromosome could replicate and produce a new set of proteins. In this experiment Mycoplasma genitalium was used. This bacterium is very small. It lives in cattle and goats. The researchers sequenced the code of its DNA and succeeded in their synthesis.

The new bacteria was called "Synthia" and will be used to create carbon-capturing algae and other useful bacteria.

Brown algae genetic code may explain origin of higher living beings

[334] Unicellular organisms originated animals, plants, fungi, red algae and brown algae. Researchers decoded the genetic code of the brown alga Ectocarpus siliculosus. This alga performs photosynthesis and is extreme versatile which are central requirements for higher life on earth.

Klaus Valentin, biologist of the Alfred-Wegener-Institute explains that scientist will choose one species of each of these five groups, and decode their genome to search for comparable genetic informations. Ectocarpus siliculosus was chosen as target in the brown algae group. The finding demonstrate that brown algae resulted from the fusion of a green alga with a red alga.

The scientists will perform further studies to see how the alga reacts to drought, UV light and rising temperatures.

The Ectocarpus genome and the evolution of brown algae [335]

Brown algae are photosynthetic organisms differ from plants in their evolution. Their genome contains 214 Millionen DNA base pairs (In molecular biology, two nucleotides on opposite complementary DNA or RNA strands that are connected via hydrogen bonds are called a base pair). Further 16.000 genes were found. Light-harvesting and pigment biosynthesis genes and new metabolic processes such as halide metabolism shows how the alga cope with the variable tidal environment. The evolution of multicellularity in this lineage is correlated with the presence of a rich array of signal transduction genes. A family of receptor kinases was found to be been linked with the emergence of multicellularity in both the animal and green plant lineages.

The Ectocarpus Genome Consortium

The Ectocarpus Genome Project started in June 2004 by an international consortium of 34 laboratories, coordinated in Roscoff, submitted a whole genome sequencing project to the French sequencing centre Genoscope. Automated annotations were obtained using the EuGene system adapted to the specificities of the Ectocarpus genome. [336]

Development and physiology of the brown alga Ectocarpus siliculosus [337]

Charrier and colleagues 2008 stress that brown algae are able to perform photosynthesis and have a cellulosic cell, but are only distantly related to plants considering the evolutionary origin of both groups. Therefore the brown algae present own features and experienced an evolution which was independent from other groups. The authors reviewed the work of the consortium of laboratories, including the Station Biologique in Roscoff and Genoscope, and two centuries of research work on Ectocarpus siliculosus.

A genetic map for the brown alga Ectocarpus siliculosus [338]

Heesch and colleagues 2010 presented A sequence-tagged genetic map for the brown alga Ectocarpus siliculosus which was proposed as a genetic and genomic model for the brown algae a genetic map using microsatellite markers that were designed based on the sequence supercontigs. The genetic map was constructed using 406 markers, resulting in 34 linkage groups. The Ectocarpus genetic map provides a large-scale assembly of the genome sequence and may be useful for further studies of the genetic of Ectocarpus siliculosus.

Phycodnaviruses may alter geochemical cycling and weather patterns [339]

Dunigan and colleagues 2006 point out that the family Phycodnaviridae consists of six genera. Their genomes range from 160 to 560kb. The phycodnaviruses have evolutionary roots that connect with several other families of large DNA viruses, referred to as the nucleocytoplasmic large DNA viruses (NCLDV).

The genome analyses have revealed more than 1000 unique genes, but only 14 homologous genes held in common among the three genera of the phycodnavirses which stands for a high gene diversity.

The authors stress the importance of phycodnaviruses infections of the phytoplankton because their effect may alter geochemical cycling and weather patterns.

Wilson, Van Etten and Allen 2009 reinforce the importance of the phycodnaviruses on infections of phytoplankton and their global global affects, on water biology. [340]

Large dsDNA viruses is integrated in the DNA of Ectocarpus siliculosus [341]

The Ectocarpus siliculosus virus-1 (EsV-1) is a lysogenic dsDNA virus belonging to the super family of nucleocytoplasmic large DNA viruses (NCLDV). It infects the brown alga Ectocarpus siliculosus. Delaroque and Bolan 2008 reports that the viral genome is integrated into the DNA of the alga.
In this study labelled EsV-1 DNA was used to identify the integration sites of the viral genome.

The authors found that some of the gene products are not encoded by EsV-1 but are present in the genome of other members of the NCLDV family, suggesting that the Ectocarpus algal genome contains traces of the integration of a large dsDNA viral genome and may be the ancestor of the extant NCLDV genomes. These viral DNA pieces might have originated EsV-1 genome through a complex integration and recombination system, using an enzyme similar to a new class of tyrosine recombinases. The authors concluded that some dsDNA viruses evolved principally through genome reduction.

Phycodnaviridae are large DNA algal viruses [342]

Van Etten and colleagues 2002 describes the genome of the Ectocarpus siliculosus virus (EsV-1) and the Paramecium bursaria chlorella virus (PBCV-1). Both are members of the family Phycodnaviridae. They infect eukaryotic algae. The 336 kb genome of EsV-1 has approximately 231 protein-encoding genes. The 331 kb genome of PBCV-1 genome has 11 tRNA genes and approximately 375 protein-encoding genes.

The two viruses have only 33 genes in common because of their different habitat. EsV-1 infects a marine brown alga, and PBCV-1 infects freshwater green algae.

Genetics of Alzheimer disease [343]

The genes CLU, PICALM, and CR1 were identified as responsible for late-onset Alzheimer disease by the Genome-wide association studies (GWAS) analysing the genetics of 35,000 persons [344]. Seshadri and colleagues 2010 report recent identification of the two novel loci rs744373 near BIN1 and rs597668 near EXOC3L2/BLOC1S3/MARK4 which, together with loci CLU and PICALM are associated with Alzheimer disease. The authors point out that both new genes are important for future research, but they are not clinically useful, because they do not improve Alzheimer disease risk prediction.

Sleegers and colleagues 2010 comment the findings of CLU, CR1 and PICALM genes which support existing hypotheses about the amyloid, lipid, chaperone and chronic inflammatory pathways in Alzheimer pathogenesis. The authors present suggestions on how these findings may improve patient care and future drug development. [345]

In another study Bettens and colleagues 2010 point out that no single functional risk variant was identified besides the three causal genes-amyloid precursor protein and presenilin 1 and 2 genes-and one risk gene apolipoprotein E (APOE), suggesting a possible involvement of rare alleles and other types of genetic variants involved in the aetiology of Alzheimer disease. [346]

Genome-wide association study [344]

A genome-wide association study is an approach that involves rapidly scanning markers across the complete sets of DNA, or genomes, of many people to find genetic variations associated with a particular disease. such as asthma, cancer, diabetes, heart disease and mental illnesses.

With the completion of the Human Genome Project in 2003 and the International HapMap Project in 2005, researchers now have a set of research tools, include computerized databases. Researchers already have reported considerable success in undestanding diseases such as age-related macular degeneration. type 2 diabetes, Parkinson's disease, heart disorders, obesity, Crohn's disease and prostate cancer, as well as genetic variations that influence response to anti-depressant medications.

Establishing a benchmark for future evolutionary modelling research in chromosome size [347]

Yu et al. 2011 tried to improve the understanding of genome and chromosome evolution which are linked to inheritance and genetic evolution. The authors compared chromosomes in different species of eukaryotes placing full genome sequences onto an X-Y axis known in cubic function calculations. [348]
This allowed them to study the genome of a species as a whole considering the evolution related to the repetition of proportions of genetic codes along chromosomes, the rules governing the similarity of the length of chromosomes in a species, the similarity of the chromosome content and the size throughout different taxonomic groups.

The researchers found that the chromosome sizes within each eukaryotic species are actually similar. Other outcomes of this research were the knowledge that the chromosomes of different organisms share a similar distribution pattern, the chromosome number of a species define the relative sizes of all the chromosomes, and the genome size in the chromosome base pair define the base pair length of each chromosome.

The majority (95 per cent) of the chromosomes in a species are expected to have a base pair length between 0.4035 and 1.8626 times the average chromosome length. Almost every genome form an S-curve of ascending chromosomal lengths at a X-Y axis, strongly suggesting a cubic function: the incremental change in chromosome size larger at both ends of the curve but smaller in the middle. Using cubic function calculation the authors found that the genome from varied species like maize, sorghum, fruit fly, dog, chimpanzee and others present the same pattern. Only genomes of platypus and birds differed from the cubic mathematical function because of their special genomes.

The authors stress that their study include empirical data analyses and computer simulations establishing a benchmark for future evolutionary modelling research in chromosome size.

Ultraviolet damage repair mechanism in plant and animals by photolyase enzyme [349]

Zhong and colleagues 2010 found that ultraviolet radiation damages the DNA by forming a 6-4 photoproduct (6-4PP) bond between two adjacent pyrimidine rings. This lesion interferes with replication and transcription, and may result in mutation and cell death. In many plants and animals a flavoenzyme called photolyase uses blue light energy to repair the 6-4PP by means of a cyclic proton and electron transfer between the enzyme and the damaged DNA. Human UV repair enzymes are less effective than those found in plants, microbes and animals. Chronic sun burns may therefore lead to skin cancer.

The researchers explain that a direct electron transfer takes place from the excited enzyme to the 6-4PP bond and a catalytic proton transfer between a histidine residue in the active site and the 6-4PP bond. After the DNA healing process, the electron and proton are returned to the photolyase enzyme which can start another reverse of the sun damage.

Genetic modified salmon may become the first GM animal at grocery stores [350]

Hearings concerning the approval of the sale of GM salmon are held in Washington, D.C. The genetic code was modified by the U.S. biotechnology company AquaBounty at Prince Edward Island. It grows twice as fast as conventional salmon. AquaBounty modified the Atlantic salmon by adding a growth hormone from a Chinook salmon and added another gene from an eel-like fish called ocean pout this allows the fish to produce their growth hormone all year long. The GM salmon needs only 16 to 18 month, while conventional Atlantic salmon needs 30 month to become full grown. The rapid growth is of high commercial interest.

According to David Senior, head of the FDA committee, there are no safety concerns regarding the GM salmon, however, present studies were performed only with a very limited number of animals.

Environmentalists argue that the GM salmon approval would also open the door for a variety of other genetically modified animals, such as the pig which is being developed in Canada or the mad cow disease resistant cattle.

A cell signaling pathway controls growth and development of certain types of cancers [351]

Some parts of gene play a regulatory role, switching the expression of the gene on or off. The gene's DNA is packaged in a substance called chromatin on which such informations may be written.

Chromatin is the combination of DNA and proteins that makes up chromosomes. The functions of chromatin are to package DNA into a smaller volume to fit in the cell, to strengthen the DNA to allow mitosis and meiosis, and to serve as a mechanism to control expression and DNA replication. Chromatin contains genetic material-instructions to direct cell functions. Changes in chromatin structure are affected by chemical modifications of histone proteins such as methylation (of DNA and proteins) and acetylation (of proteins), and by non-histone, DNA-binding proteins. [352]

The instructions laid down on the chromatin may be "bookmarks," substances placed on the chromatin by so-called epigenetic "writer" proteins. Another group of epigenetic proteins, known as "erasers," are able to remove the bookmarks. Both types of proteins have successfully been disabled by scientists, using molecules made in the lab or taken from nature.
A third variety of epigenetic proteins switch genes on or off by "reading" the bookmarks.

Teitell and colleagues 2010 describe a new signaling pathway which is part of a global DNA damage response controlling 136 genes which cause cancers, such as lymphoma and leukemia.

The 136 genes of the signaling pathway are turned off when less mature cells develop to mature cells. Defects in this pathway, however, don't allow cells to mature the signaling genes are not silenced. The immature cells begin to divide rapidly resulting in lymphoma or leukemia.

B-cells produce antibodies to fighting an infection or react to other disorders. This results in DNA damage of the B-cells activates the gene TCL1 of the signaling pathway and causes cancer, if the pathway is not turned off. Activation of the signaling pathway silences the TCL1 gene and cancerous cell are not formed.

During an immune response, B cells undergo rapid proliferation and remodeling of genes producing immunoglobulin (IG) genes within germinal centers (GCs) to generate memory B and plasma cells. Unfortunately, the genotoxic stress associated with the reaction of the germinal centres may promotes cancerous B cells.

The authors explain that the body naturally turns off the signaling pathway after about two to three weeks after a DNA stress, however, if the pathway gets defect the natural stop signal is interrupted and cancerous cells develop. Targeting this pathway cancer therapies could be enhanced, say the authors.

Optogenetics

Optogenetics is an emerging field combining optical and genetic techniques to study neuronal circuits in live animal at the high speeds (millisecond-timescale)within intact mammals and other animals. Optogenetics uses light-activated channels and enzymes that allow manipulation of neural activity with millisecond precision while maintaining cell-type resolution in the brains of intact animals. This allows to study the causal impact of biochemical signalling in behaving mammals, in a targetable and temporally precise manner. In comparison, traditional genetics targets "on" or "of" function changes in cell proteins, to understand their function.

Genetically targeted photostimulation methods rely on the use of opsins and a variety of technical achievments tried to attain photonic control of neuronal firing, later several microbial opsins were introduced, such as Channelrhodopsin-2, a single-component light-activated cation channel from algae. ChR2, related microbial proteins and customized conditionally-active viruses are used to introduce the light-sensitive particles to neurons in the brain of living animals. These proteins are used to control the activity of neurons avoiding several of the non-specific effects of electrical or pharmacological brain stimulation. [353]

Microbial opsins being used are cation channels such as Channelrhodopsins ChR2 and VChR1 to excite neurons, and chloride pump such as Halorhodopsin (NpHR), Archaerhodopsin (Arch), and fungal opsins such as Leptosphaeria maculans opsin (Mac) to inhibit neurons. [354]

G-protein coupled receptors are linked vertebrate opsins which can target intracellular messengers such as cGMP, cAMP and IP3 [355]. Stierl et al 2011 propose photoactivated adenylate cyclases (PACs) from Euglena and marine bacteria to control cAMP. Expressed in the fly nervous system, [356]

Kokaia and Soerensen 2011 that microbial opsonin genes channelrhodopsin-2 (ChR2) may be used as light- activated ion channels and halorhodopsin (NpHR) as pumps when exposed to light and trigger action in ChR2-expressing neurons, and inhibition in NpHR-expressing neurons. The authors stress the possibility to restore the dopamine-related movement system in parkinsonian animals, amelioration of blindness and recovery of breathing after spinal cord injury are a few examples of such perspectives. [357]

Study on similarities in genomes across species provide better understanding of inheritance and evolution

[358] The project leaded by Jianming Yu, 2011 [1], compared 886 chromosomes in 68 random species of eukaryotes, placing each fully sequenced genome onto an X-Y axis. Trying to understand genetic inheritance and evolution the authors studied the genome of a species as a whole, addressing the evolution and variation of individual chromosomes across species, such as the repetition of proportions of genetic codes along chromosomes, the rules governing the similarity of the length of chromosomes in a species, and similarity of the chromosome content and size throughout different taxonomic groups.

The researchers found that the chromosome sizes within each eukaryotic species are actually similar rather than drastically different as previously believed. They also found that the chromosomes of these different organisms share a similar distribution pattern. The chromosome number of a species define the relative sizes of all the chromosomes, and the genome size in the chromosome base pair define the base pair length of each chromosome. The majority of the chromosomes in a species are expected to have a base pair length between 0.4035 and 1.8626 times the average chromosome length. Almost every genome form an S-curve of ascending chromosomal lengths at a X-Y axis. The genome from varied species like maize, sorghum, fruit fly, dog, chimpanzee and others present the same pattern. Only genomes of platypus and birds deviated from forming an S-curve because of the particularities of these genomes. The distribution of the genomes are dictated by the laws of mitosis, meiosis and cell division, according to findings of Guihua Bai co-author of the study.

Salt stress and tetrapyrrole metabolism [359]

The translocator protein 18 kDa (TSPO) is important for many cellular functions in mammals and bacteria, such as steroid biosynthesis, cellular respiration, cell proliferation, apoptosis, immunomodulation, transport of porphyrins and anions. TSPO primarily functions to transport heme, porphyrins, steroids and anions.

TSPO proteins are also important for cellular respiration, cell proliferation and apoptosis.Arabidopsis thaliana contains a single TSPO/PBR-related gene with a 40 amino acid N-terminal extension it might be chloroplast or mitochondrial localized.

Balsemão-Pires et al.2011 report that AtTSPO is involved in the response of Arabidopsis to high salt stress. Salt stress leads to re-localization of the AtTSPO from the endoplasmic reticulum to chloroplasts through its N-terminal extension. The authors propose that AtTSPO may play a role in transporting tetrapyrrole intermediates during salt stress and other conditions in which tetrapyrrole metabolism is compromised.


Tetrapyroles

Higher plants synthesize four major tetrapyrroles (chlorophyll, haem, sirohaem and phytochromobilin). In metazoans, heme and siroheme are synthesized in mitochondria, but in plants tetrapyrrole biosynthesis is plastid-localized, suggesting that tetrapyrroles are transported from the chloroplast to the mitochondria.

Accordimg to Balsemão-Pires and colleagues, the expression of PpTSPO-1 is induced by salt stress in Physcomitrella patens, and also in Arabidopsis, Plant TSPO has been proposed to participate in the interaction between plastid and mitochondrial tetrapyrrole biosynthetic pathways. In higher plants, tetrapyrroles are synthesized almost exclusively in plastids, with the exception of the two last steps of heme synthesis that may occur in both chloroplasts and mitochondria.

Vitamin E and tannic acid reduces DNA damage induced by alcohol [360]

Jordão et al 2011 assessed the DNA protection role of vitamin E and tannic acid during oxidative stress induced by alcohol in rats for four weeks of ethanol containing diet. Chronic ethanol consumption causes lipid peroxidation by increasing free radicals or reducing antioxidants. This may damage hepatic DNA. The authors measured plasma vitamin E and liver glutathione, thiobarbituric acid resistant substances, and a-tocopherol, together with the hepatocytes comet test.

DNA damage caused by ethanol during a period of 24 hours, was absent in rats receiving tannic acid or vitamin E. Tannic acid was most effective in reducing steatosis, a fatty degeneration of cells. The authors call for a reevaluation of the antioxidant role of vitamin E and of tannic acid, with special focus on the protective role of tannic acid against hepatic steatosis.


The hepatocytes comet test

According to Ashby et al 1995, the main liver cells, called hepatocytes, present a comet tail at a gel electrophoresis assay, when pretreated with DNA damaging substances. The authors describe the methodology and the measurement of the comet tail enabling to assess ongoing cell damages. [361]

The comet assay measures DNA strand breaks with higher sensitivity as similar tests, such as sister chromatid exchange, alkali elution and micronucleus assay. Lee and Steinert write that the comet assay is often used to assess the activity of genotoxicants on aquatic animals. The authors suggest to use both the intensity of the tail and the length of the tail. This means to use the DNA tail moment, percentage of DNA in the tail. The evaluation of the comet tail should be standardised to give comparable results. [362]

CRISPR, a RNA-based immune systems of bacteria

Clustered regularly interspaced short palindromic repeat (CRISPR) are nucleic-acid-based adaptive immune systems in bacteria and archaea. These small RNAs (crRNAs) in bacteria find and silence invading foreign nucleic acids, such as viruses and plasmids. Wiedenheft et al. 2012 point to the similarity to the RNA interference (RNAi) pathways in eukaryotes but differences in the silencing must be further studied. [363]

Short sequence tags from invading genetic elements are actively incorporated into the CRISPR locus of the attacked bacteria. The invading DNA is then transcribed and processed into a set of small RNAs leading to the destruction of the foreign genetic material. [364]
CRISPR sequences provide an adaptive, heritable record of past infections and express CRISPR RNAs - small RNAs that target invasive nucleic acids. [365]

Jinek et al 2012 describes the adaptive immunity against viruses and plasmids of bacteria based on CRISPR/Cas systems which uses crRNAs to silencing invading nucleic acids. In this system a CRISPR-associated protein Cas9 is brought to introduce double-stranded (ds) breaks in invading DNA. At sites complementary to the crRNA-guide sequence, the Cas9 HNH nuclease domain cleaves the complementary strand while the Cas9 RuvC-like domain cleaves the noncomplementary strand. The study sheds light into a family of endonucleases that use dual RNAs for site-specific DNA cleavage. [366]

Almost all archaea and approximately half of bacteria hold CRISPR-Cas systems writes Bhaya et al.2011. The CRISPR-Cas immune systems capture short invader sequences within the CRISPR loci in their genomes, and small RNAs produced from the CRISPR loci (CRISPR (cr)RNAs) guide Cas proteins to recognize and degrade or silence the invading nucleic acids. According to Bhaya, the CRISPR-Cas pathway functions in three phases adaptation of CRISPRs to invaders, crRNA biogenesis, and invader silencing. [367]

The CRISPR RNAs (crRNAs) contain sequence elements acquired from invaders that guide CRISPR-associated (Cas) proteins back to the complementary invading DNA or RNA. The crRNAs are associated with the Cas RAMP module (Cmr) effector complex, which cleaves targeted invasive RNAs. Hale et al. 2012 found that the Cmr crRNAs contain an 8 nucleotide 5' sequence tag that is critical for crRNA function. It can be used to engineer crRNAs that direct cleavage of novel targets. The authors suggest that the CRISPR RNA-Cmr protein pathway may be used to cleave RNAs of interest. [368]

The CRISPR/Cas system builds inheritable DNA-encoded immunity following loger exposure to invasive DNAs, however, viruses create mutational escape strategies that allow them to evade the CRISPR/Cas system. [369] Safer Escherichia coli using transfer of CRISPR/Cas adaptive immune system [370] Sapranauskas et al 2011 found that the CRISPR/Cas adaptive immune system of Streptococcus thermophilus can be transferred across distant genera, llike Escherichia coli. Strains of Escherichia coli may be turned more resistant to phage attack, and less prone to uptake and disseminate plasmid-encoded undesirable genetic elements.

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