Dr. Mae-Wan Ho Replies to Aventis in Chardon LL Public Hearing
10 June 2002, London
Aventis applied to place herbicide-tolerant transgenic maize Chardon LL on UK National Seed List in 2000. This provoked such widespread objection that a public hearing was held later that year. But the hearing was adjourned when it transpired that Chardon LL did not pass the requisite test for distinctness, uniformity and stability (DUS). The hearing was resumed in May 2002, when Aventis made an extended oral submission. Dr. Mae-Wan Ho replies to claims of Aventis on the genetic stability of Chardon LL, the CaMV 35 S promoter, and horizontal gene transfer. She was speaking on behalf of The Burnhams Group.
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MR AYLESBURY: I think I understood you to say that you are going back there and then Dr Mae-Wan Ho. That is fine, if you want to -- yes, that is the advantage of going there, I quite agree. It is the best place, really. Now, Dr Ho, welcome back to the hearing. I had already been given BG 33, 34 and 35, which are documents relating to what you are going to say. I think 33 was what I understood to be the nearest thing to a text for what you are going to say. I have also just been handed 37 and 38, which were the documents sent out for copying just a moment ago. No doubt you will, as you go through, tell me what they are and what I need to understand about them.
DR. HO: Yes.
MR. AYLESBURY: If you would like to proceed, then.
DR. HO: Okay. Thank you very much for the opportunity, everyone, for coming here and for letting me speak again. I also would like to echo Dr Elliott's comments of thanks and appreciation to yourself and to your staff.
Now, what I would like to do is to comment mainly on points raised in Aventis’s oral submission made on the 29th May and specifically to the paragraphs 10.4 to10.16, which Mr Aylesbury considers to be new evidence countering alleged instability of Chardon LL; and to paragraphs 10.19 to 10.20, which concern evidence on the cauliflower mosaic virus or CaMV 35S promoter that I had presented to this hearing earlier.
I would then like to take this opportunity to update on some of the evidence against the proposed addition of Chardon LL to the UK National List, which has emerged since the hearing was adjourned, especially on horizontal gene transfer.
First of all, the point I would like to make is thatthe stability of Chardon LL is still unproven. Paragraphs 10.4 to 10.16 of Aventis's document purport to describe the process whereby event T25 containing the genetic modification that produces the enzyme phosphinothricin acetyltransferace, or PAT for short, in maize was selected from the primary transformants, resulting in an inbred T25 line, which was then repeatedly back crossed to the non-GM variety for a number of generations. Now, exactly how the crosses were made was not stated. This is very important, if you really, really want to know whether it ended up inbred or not.
There are numerous assertions throughout the paragraphs that the line or lines are stable, for which no evidence whatsoever was presented. There are no data to support the stability of the original T25 so called inbred line. It is not even stated how many generations that line has been perpetrated. As long as this inbred line and the subsequent repeatedly back crossed line referred to as converted inbred require selection with the herbicide glufosinate ammonium, or GA for short, then the lines are clearly either not uniform or not stable, or both: otherwise, practically all the plants would be expected to be GA tolerant.
Paragraph 10.7 states:
"During every generation of the back cross process, GA was sprayed before flowering to identify the segregating plants carrying the GA tolerance, and to ensure that all plants not expressing the trait could not contribute to the progeny."
However, without counting the numbers of the plants that are GA tolerant and GA susceptible and without performing the necessary molecular analysis, it is not possible to distinguish so called segregating plants which are progeny derived from the non-GM plants, that is those that never had the transgene at all, from the GM plants that have simply gone unstable.
Paragraph 10.10 states:
"Since the last back crossing step, the converted inbred had undergone at least five generations of self-crossing, without giving reason to doubt the line’s lack of expression or stability. In all multiplication steps of the T25 line, GA was applied before flowering so any plants not expressing the trait would not contribute to the progeny. The frequency of "off types" (looking at their more following or lack of expression of the GA tolerance gene) was very low, especially as compared to the norm in the seed industry for pre-basic and basic seed."
But if the back cross had really generated a so called converted inbred line, there should be no GA susceptible plants remaining, if the line were uniform and stable, again. And without numerical and molecular data, we cannot know if the line fell within the accepted norm or not.
More to the point, stable expression of the trait is no guarantee of genetic stability. To prove genetic stability requires characterisation by molecular analyses. This is called event specific molecular analysis. It is to demonstrate that both the structure and location of the transgenic insert or inserts remain unchanged in successive generations. No such data have been presented.
Towards the end of last year, the GA tolerant oilseed rape crop, planted as part of the farm scale evaluation in Black Isle near Munlochy, suffered extensive herbicide damage after it was sprayed with glufosinate ammonium. I have a picture here in BG 38.
On page 30, you can all see on one side the yellow stunted crop, which is the GM crop after it was sprayed with the herbicide glufosinate ammonium. This other side is the non-GM crop.
MR. AYLESBURY: Yes, it is not so obvious in the photocopy but yes.
DR. HO: And that was a sign of massive transgenic instability in another crop, GA tolerant crop produced by the same company, Aventis. The company has yet been unable to offer a counter explanation.
The new European directive already referred to for deliberate release, 2001/18/EC, specifically requires molecular data documenting that the GMO is genetically stable.
So in order to prove that the lines are stable as claimed, Aventis must now present all past and present molecular data and provide samples for independent analysis.
The second point I would like to make is that the CaMV 35S promoter is unstable. Paragraphs 10.19 to 10.20 attempt to discredit the evidence on the CaMV 35S promoter I had submitted to the hearing, which consisted at least of three papers I and my colleagues published in a scientific journal.
However, Aventis referred to the first paper only, but not the other two that had answered all scientific criticisms raised, as I pointed out in my previous submissions, and I will not repeat them here.
Instead of dealing with the scientific criticisms, the company has chosen to cite copiously from a so-called "analysis" by a science journalist, not a scientist, and it was published under "Business and regulatory news" in the journal Nature Biotechnology.
That so-called "analysis" was concocted entirely of hearsay and opinions, and contained such defamatory, and libellous statements, that the journal had to give us a right to reply when challenged. Our reply was published, along with the journalist's so-called "apology" that he had failed to cite our rebuttal, but he attacked us yet again. This time, however, Nature Biotechnology refused to print our reply, which appeared elsewhere subsequently; and I brought this to the last hearing, this is BG35, but at that time it was brought in the proof, galley proof form. This is now in the published form.
MR. AYLESBURY: Yes, thank you.
DR. HO: Now, our claims that, (quotes from Aventis’ Submission), "CaMV promoter may be structurally unstable and prone to horizontal gene transfer and recombination" are based on extensive reviews of existing scientific findings, some of which emerged long after the promoter had been widely used. A key evidence that provoked us to write the review was the discovery of a "re-combination hotspot" in the promoter by at least two groups of researchers. In the course of our debate, we also discovered that the promoter is active in species across the entire living world, including human cells, and this was what the subject of BG35 was about.
Now, our critics appeared not to know that. Aventis too was ignorant, as were two experts invited to the recent public hearing conducted by ACRE. This report has been circulated by the institute that I direct, which is Institute of Science and Society via the E-Mail, it is available on our website, www.i-sis.org.uk, and it will also be appearing in the upcoming issue of Science in Society, the official publication of our institute.
Now, two years later, and this is very significant, one of the research groups that had discovered the CaMV 35S promoter recombination hotspot called for this promoter to be phased out, and this appeared in the internal report of the John Innes Centre, and the reason given was because of the instability it causes. But structural instability would indeed make this CaMV promoter prone to horizontal gene transfer and re-combination as we have pointed out.
Last November, again this is new evidence, a paper published in Nature reported that Mexican maize landraces growing in remote regions have been contaminated by transgenic maize. The paper is now at the centre of a storm whipped up by scientists supporting biotech, who are criticising it for so-called "poor methodology". This led to a retraction by Nature, which is unprecedented for a paper that has not been proved wrong or fraudulent. The critics are not contesting the fact that the transgenic contamination has occurred and that the CaMV 35S promoter has been found in the landraces. In fact, the Berkeley researchers who wrote the original report were able to present new data confirming, firming up their conclusion that transgenic contamination in the form of a CaMV 35S promoter had occurred. What their critics are contesting is the form in which the promoter has entered the landraces’ genomes. Again, you can read the details in reports we have already circulated and again in the upcoming issue of Science in Society. This is BG37.
The researchers claimed to have found that the promoter in the landraces’ genomes is linked, not to the original transgenes, but to a variety of other DNA sequences. That is as though the promoter has broken off and joined up at random, or as stated by one of the critiques published in Nature, "fragmenting and promiscuously scattering through the genomes". This, the critics are strenuously denying.
But that is far from unexpected, in view of the "recombination hotspot" which is after all a fragmentation hotspot associated with the35S promoter, CaMV 35S promoter, which was discovered years after the transgenic crops containing the promoter have been widely released. Crucial molecular data that could settle this dispute either do not exist, or are hidden under so-called "commercial confidentiality". Three companies, Monsanto, Novartis and Aventis have all created transgenic maize containing the CaMV 35S promoter. But without the "event-specific" molecular data required it is difficult to find out which of the transgenic maize has contaminated the landraces.
I repeat, the companies must now provide both past and present molecular data on their transgenic lines and submit samples for independent analysis. It is of interest that when Monsanto's Roundup Ready was subjected to such analysis, the transgenic insert was found to be scrambled up, as was the host genome at the site of insertion. And a DNA fragment of more than 500 basepairs of unknown origin is also present. If you do not know where it came from, you do not know whether it is safe or not. None of that was reported in the company's original applications submitted for commercial approval.
One further piece of new evidence that I would like to refer to is horizontal gene transfer. I have circulated, and I have brought a report that was recently circulated on our email list and is also up on our website. It is called "Denial Continues Over Horizontal Gene Transfer". This is I believe BG34.
MR. AYLESBURY: BG34, yes.
DR. HO: That is right. I will not go through it in detail.
MR. AYLESBURY: Right.
DR. HO: But just to point out that there has been a lot of selective citing of negative evidence, and also explaining away of positive incriminating evidence of horizontal gene transfer. It is even more ridiculous than that. Positive evidence is dismissed by an oxymoronic so-called cautious interpretation that eschews any and all positive results. So on our website we have plenty of evidence gathered together on horizontal gene transfer and in many cases you will find that unless you read the paper carefully, if you just read the title or the extract, you will think that it is negative result. But that is not the case when you actually read the report in detail as I have had to for punishment I think. It is really very boring to read these reports.
So there is definitely evidence suggesting that in the only field study carried out so far that transgenic DNA not only persists in the soil for two years after the GM crop has been harvested. Parts of the transgenic DNA were found to have transferred to soil bacteria in batch cultures, there were positive results identified, even though the actual strain of bacteria cannot be isolated and that is not surprising because less than 1 per cent of the bacteria in the environment can be cultured.
Then the question arose as to whether transgenic DNA, or DNA can get into mammalian cells. In my previous submission I have already mentioned that from the many, many experiments done on so-called "gene therapy", so-called because nobody has been cured yet, there is no doubt that DNA, transgenic DNA can get into mammalian cells. However, there is a research team in Cologne, in Germany, that began the first investigations as to whether transgenic DNA and viral DNA can get into cells. I already summarised their findings when I was here last time. Basically when they fed mice with bacterial virus M13 DNA, or the cloned gene for the green fluorescent protein inserted into a plasmid, they found that large fragments of the DNA can be traced from the intestinal contents, via the gut wall, the Peyer’s patches, those are in the gut wall, and white blood cells to the spleen and the liver. When fed to pregnant mice the DNA passed via the placenta to foetuses and newborn animals.
Now, in a recent experiment the same team fed ordinary non-transgenic soya leaves to mice in order to find if the natural gene of the soya could be transferred to the mice, to the cells of the mice, at the same time they also looked at whether transgenic DNA containing the green fluorescent protein linked to some viral promoter, several viral promoters, could also be transferred.
Now, they found that indeed, in confirming their early results, the transgenic DNA was very readily transferred to a large number of cells just 3 to 8 hours after the feeding. Whereas in the soya DNA there were very few positives, relatively few positives. However, as they point out, the conditions are not equivalent because the amount of DNA fed as transgenic DNA was much larger than the soya DNA. So you know one would ask the obvious question: why did they not do the obvious experiment, which is to feed transgenic plant material to the mice and probe for both a plant gene and the transgene? And that would have settled the dispute, because the main question is: are there any reasons to suspect that transgenic DNA is more frequently, more readily transferred, than non-transgenic? I have given reasons in my earlier submission why I think there are very good reasons to believe that is the case.
Unfortunately this obvious experiment has never been done perhaps because biotech companies will not provide transgenic material for such experiments. In fact this obvious experiment appears to have been avoided altogether, so the denial can continue because after all absence of evidence has been copiously taken to be evidence of absence.
So now to summarise. The new evidence that has emerged has only strengthened my objections. I remain strongly opposed to placing Chardon LL on the National Seed List. Thank you very much.
MR. AYLESBURY: Thank you very much indeed, Dr Mae-Wan Ho. Thank you for that. Good. I presume, Mr Smith, you are not at this late stage in the proceedings going to be taking up cross-examination opportunities or anything like that. No, good.
The Written version with references is now available on the ISIS members website. Membership details are available here.
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