Healthy soil and sustainable growing

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The Best Kept Secret of GM Crops
Witness Statement to ACRE
http://www.i-sis.org.uk/secretGMcrops.php
For ACRE open hearing on the criticisms of T25 GM maize risk assessment

The hearing will take place from 10.00am to 2.00pm on Wednesday, 20
February, in Room 7A, B and C, Ashdown House, Department for Environment, 
Food and Rural Affairs, 123 Victoria Street, London SW1E 6DE.

Dr. Mae-Wan Ho

Institute of Science in Society, PO Box 32097, London NW1 0XR, UK

I am speaking against the market approval of T25 because there is no evidence 
that it is a genetically stable, uniform line, the single most important 
criterion for approval. For unless it is genetically stable, you might as 
well forget about environmental or health risk assessment. And genetic 
instability is also a serious safety issue. The public hearing on T25 was 
suspended over a year ago when it was found not to have passed the required 
EC test for Distinctiveness, Uniformity and Stability (the DUS test), as I 
pointed out when giving evidence to the hearing [1].

The new EC Directive on deliberate release requires strict molecular evidence 
of genetic stability, which is also necessary for establishing the identity 
of the transgenic line and to ensure traceability. The best-kept secret of GM 
crops is that they are not stable.

There is a large literature on gene silencing, in which the transgenes remain 
in the genome, but are not expressed. More serious, from the safety point of 
view, is structural instability, the tendency for the transgenic DNA to come 
loose, to rearrange or become lost in part or in whole in successive 
generations [2,3]. This could change the transgenic line in unpredictable 
ways in terms of health and environmental risks. And it will increase the 
chance of transgenic DNA being taken up by unrelated species to make new 
combinations with their genetic material. That’s referred to as horizontal 
gene transfer and recombination. Transgenic DNA can spread to every species 
that interact with the transgenic plant, in the soil, in the air, in the 
mouth and gut and the respiratory tracts of animals including human beings.

New viruses and bacteria that cause diseases could be generated, and 
antibiotic resistance marker genes could spread to the pathogens. Transgenic 
DNA may also get into human cells and insert into the human genome; and a 
large body of evidence from so-called gene therapy experiments have amply 
demonstrated this does occur [4]. The constructs used in gene therapy are 
very similar to those used in transgenic plants, and one main side-effect of 
transgenic DNA inserting into human genome during gene therapy is cancer.

Despite that, our regulators have not required biotech companies to provide 
molecular evidence of stability. ACRE’s latest guidelines for industry put 
out for public consultation asks industry to provide molecular evidence of 
genetic stability over one generation only [5], which is derisory. We need 
data for at least five successive generations [6]. No such data have come 
forward from the companies. On the contrary, companies have been allowed to 
hide under ‘commercial confidentiality’.

I am putting to you twelve reasons why trangenic DNA is different from 
natural DNA, and is more likely to spread by horizontal gene transfer and 
recombination, both by design and otherwise. I hope you will refute these 
point by point.

(The details are in two ISIS reprint collections on transgenic instability 
and horizontal gene transfer that I am presenting to ACRE, for free.)

    * All artificial constructs tend to be unstable, so much so that this is 
a topic in a standard text-book on genetic engineering [7]. Transgenic DNA is 
more likely to break and join up again, ie, to recombine.
    * Transgenic DNA typically contains DNA from widely different sources, 
mainly bacteria and viruses and other genetic parasites that cause diseases 
and spread antibiotic resistance, and hence, has the potential to recombine 
homologously with all those agents, ie, due to similarities in DNA 
base-sequence. Homology enhances horizontal gene transfer 10 million to 100 
million-fold [8].
    * Transgenic DNA is designed to cross species barriers and to invade 
genomes. They are flanked by recombination sequences, such as the left and 
right borders of T-DNA or the terminal repeats of viral vectors, which enable 
them to jump into genomes. By the same token, they could jump out again. 
Enzymes catalysing jumping in also catalyse jumping out.
    * Certain ‘receptive hotspots’ have now been identified in both the plant 
[9] and the human genome [10]. These may also be ‘recombination hotspots’, 
prone to breaking and rejoining. That would mean inserted transgenes are more 
likely to be lost, to recombine, or to invade other genomes.
    * There are mechanisms in the cell that actively seek out, inactivate or 
eliminate foreign DNA from the genome [11].
    * Cell and embryo culture methods are well-known to induce unpredictable, 
uncontrollable (somaclonal) variations that persist in the plants generated. 
There is now evidence that the transformation process for making transgenic 
plants induces further genetic instability [12-14] leading to chromosomal 
rearrangements, genome scrambling, in other words.

    * Monsanto’s Roundup Ready soya, commercially grown for years, was 
finally analysed by molecular methods. Not only is the gene order of the 
insert found to be scrambled, the plant genome at the site of insertion is 
also scrambled, and there is a 534 bp fragment of unknown origin in there as 
well [15]. All very different from the original data provided by Monsanto.

    * Recombination hotspots within the transgenic DNA, such as that 
associated with the ubiquitous cauliflower mosaic virus (CaMV) 35S promoter, 
could enhance horizontal gene transfer and recombination. We highlighted that 
in 1999 [16-18], and demanded that all transgenic crops with the promoter 
should be immediately withdrawn for safety reasons. Two years later, the 
researchers who discovered the promoter’s recombination hotspot also 
recommended that it should no longer be used [19], not because of safety, but 
because its instability compromises agronomic performance.

    * Recently, landraces of corn growing in remote regions of Mexico were 
found contaminated with transgenic corn DNA by probing with the CaMV 35S 
promoter [20]. Molecular analysis showed that the sequences next to the 
promoter are very diverse, as consistent with horizontal gene transfer and 
recombination [21].
    * CaMV 35S promoter is active in species across the entire living world, 
including frog eggs and human cells [18], as we uncovered in the literature 
more than ten years old that had apparently escaped the notice of plant 
geneticists who attacked us. CaMV 35S promoter, if transferred to human or 
animal cells, could activate cancer-associated genes as well as dormant 
viruses that are in all genomes. Another side effect of gene-therapy is the 
generation of active viruses in cell lines used to package the gene-therapy 
vectors [4]. Our critics are still dismissing the risks of CaMV 35S promoter, 
but are avoiding doing any experiments. It is a case of don’t look, don’t see 
[5].
    * Transgenic DNA from GM plants was found to transfer to soil bacteria. 
The possibility of transfer to bacteria in the mouth and gut of animals was 
suggested in laboratory investigations funded by the UK government. There is 
also evidence suggesting that transgenic DNA from crop plants has transferred 
to soil bacteria in the field [22]. But ACRE has ignored that by a selective 
interpretation of the scientific evidence that seems to me contrary to both 
the precautionary principle and good science [23].

In summary, there is no reason to believe T25 is stable. Furthermore, it has 
especially hazardous sequences, including the CaMV 35S promoter and an 
ampicillin resistance gene that, though inactive, can easily be transferred 
into integrons that will provide it with a promoter to make it functional 
[1]. T25 has uncharacterised sequences that might be involved in causing 
diseases. Finally, it has an origin of replication, which enables the 
transgenic DNA to be replicated as a plasmid if transferred into bacteria, 
thereby greatly increasing horizontal gene transfer on to other species. The 
origin of replication is also a recombination hotspot, and there have been 
strong recommendations from a recent joint FAO/WHO Expert Consultation on 
Foods Derived from Biotechnology that transgenic lines containing this 
sequence should not be approved on safety grounds [24].

   1. Ho MW. Chardon LL Public Hearing Ocober 26 2000 on behalf of Burnham 
Group, also in transcript.
   2. See Ho MW. Genetic Engineering Dream or Nightmare? Gateway, Gill & 
Macmillan, Bath and Dublin, 1998, 1999, Chapter on Perils amid Promises of 
Genetically Engineered Foods.
   3. ISIS Reprints on Transgenic Instability, 1999-2001, ISIS Publications, 
London.
   4. Ho MW, Ryan A, Cummins J and Traavik T. Slipping Through the Regulatory 
Net: ‘Naked’ and ‘Free’ Nucleic Acids, Third World Network Biotechnology 
Series, Third World Network, Penang 2001.
   5. See Watering down EC Directive on Deliberate Release ISIS Report, 
February 2002.
   6. Ho MW and Steinbrecher RA. Fatal flaws in food safety assessment: 
critique of the joint FAO/WHO Biotechnology and Food Safety Report. 
Environmental & Nutritional Interactions 1998, 2, 51-84.
   7. ISIS Reprints on Horizontal Gene Transfer, 1999-2001, ISIS 
Publications, London.
   8. Principles of gene manipulation, by Old and Primrose, Blackwell 
Science, 5th ed, 1994.
   9. DeVries J, Meier P and Wackernagel W. The natural transformation of the 
soil bacteria Pseudomonas stutzeri and Acinetobacter sp. by transgenic plant 
DNA strictly depends on homologous sequences in the recipient cells. FEMS 
Microbiology Letters 2001, 195, 211-5.
  10. Kumar S and Fladung M. 2000. Transgene repeats in aspen: molecular 
characterisation suggests simultaneous integration of independent T-DNAs into 
receptive hotspots in the host genome. Mol Gen. Gent 2000, 264, 20-8.
  11. Miller DG, Rutledge EA and Russell DW. Chromosomal effects of 
adeno-associated virus vector integration. Nature genetics 2002, 30, 147-8.
  12. Kumpatla, S.P., Chandrasekharan, M.B., Iyer, L.M., Li, G. and Hall, 
T.C. (1998). Genome intruder scanning and modulation systems and transgene 
silencing. Trends in Plant Sciences 3, 96-104.
  13. Horvath H, Jensen L,Wong O, Kohl E, Ullrich S, Cochran J, Kannangara C, 
and von Wettstein D. Stability of transgene expression, field performance and 
recombination breeding of transformed barley lines, Theor Appl Genet. 
2001,1-11.
  14. Svitashev S, Ananiev E, Pawlowski WP, and Somers DA. 2000. Association 
of transgene integration sites with chromosome rearrangements in hexaploid 
oat. Theoretical and Applied Genetics 2000, 100,: 872-80.
  15. Tax FE and Vernon DM. T-DNA-associated duplication/transloations in 
Arabidopsis. Implications for mutant nanalysis and functional genomics. Plant 
Physiology 2001, 126, 1527-38.
  16. Windels P, Taverniers I, Depicker A, Van Bockstaele E and De Loose M 
(2001). Characterisation of the Roundup Ready soybean insert. Eur Food Res 
Technol DOI 10.1007/ s002170100336, © Springer-Verlag; see also "Scrambled 
genome of Roundup Ready soya" by Mae-Wan Ho, ISIS Reprints on Transgenic 
Instability, 1999-2001, ISIS Publications, London.
  17. Ho MW, Ryan A and Cummins J. Cauliflower mosaic viral promoter – a 
recipe for Disaster? Microbial Ecology in Health and Disease 1999: 11: 
194-197.
  18. Ho MW, Ryan A and Cummins J. Hazards of transgenic plants with the 
cauliflower mosaic viral promoter. Microbial Ecology in Health and Disease 
2000: 12: 6-11.
  19. Ho MW, Ryan A and Cummins J. CaMV35S promoter fragmentation hotspot 
confirmed and it is active in animals. Microbial Ecology in Health and 
Disease 2000: 12: 189.
  20. Christou P, Kohli A, Stoger E, Twyman RM, Agrawal P, Gu X. Xiong J, 
Wegel E, Keen D, Tuck H, Wright M, Abranches R and Shaw P. Transgenic plants: 
a tool for fundamental genomics research. John Innes Centre & Sainsbury 
Laboratory Annual Report 1999/2000, p. 29. See "Top research centre admits GM 
failure" ISIS Reprints on Transgenic Instability, 1999-2001, ISIS 
Publications, London.
  21. Quist D and Chapela IH. Transgenic DNA introgressed into traditional 
maize landraces in Oaxaca, Mexico. Nature 2001, 414, 541-3, 2001.
  22. "Transgenic pollution by horizontal gene transfer?" by Mae-Wan Ho, in 
ISIS Reprints on Horizontal Gene Transfer, 1999-2001, ISIS Publications, 
London.
  23. "Horizontal gene transfer happens. A practical exercise in applying the 
precautionary principle" by Mae Wan Ho in ISIS Reprints on Horizontal Gene 
Transfer, 1999-2001, ISIS Publications, London.
  24. Joint FAO/WHO Expert Consultation on Foods Derived from Biotechnology, 
WHO Headquarter, Geneva, September 24-28, 2001.