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HomeMy WebLinkAboutCOM 0271.047 2012-2014 • PALMS PACIFIC AGRICULTURE LAND MANAGEMENT SERVICES COUNTY CLERK May 14,2013 COUNTY OF HAWAI'I CEIVED To: Hawaii County Council Time(IV q_, By Date b-/0-3 Re: Bill 97 Greetings to the Council and thank you for hearing my views.I have come before the Council previously on matters similar to this as a Geneticist(Ph.D.1974,UHM)and as Dean of the College of Agriculture,Forestry and Natural Resource Management at UHH before my retirement and now as a private business owner dealing with agricultural issues. In previous statements I brought before the Council scientific facts in attempts to avoid the emotional aspects that so often go along with testimony regarding the subject of allowing genetically modified organisms(GMOs)into the community.All over the country,these issues exist in serious discussion of what to do,and those who parley GMO seeds and plants for profit are the most outspoken supporters for them.Too often they point to the"good things"these will do for humanity in terms of food security and growth and job creation but NOT to their own tests of these organisms in their laboratories.Strangely this information is not being released to the public on grounds it is the basis of their corporate patents and could be stolen. The view(from the public's end)is not transparent which should concern representatives such as yourselves. However,even though attempts have been made to suppress them,individual scientists have stepped up to conduct investigations.As I pointed out some years ago before this body,these independent investigations would someday reveal the real problems that GMOs can bring to our communities.This prediction is bearing true. More and more studies are bringing to the open scientific literature the threats posed by GMOS.The most startling of these is our worst fear;that the genetically modified gene sequences might be able to escape the genomes they are inserted into and spread to other organisms. I am attaching to this document some of these recent studies.Please pay special attention to Dr. Nina Federov's Presidential Address to the Association for Advancement of American Science(AAAS),one of the leading scientific bodies in the USA. Her speech on transposons highlights the fact that these genetic"jumping genes"can move from one part of the DNA to another and even between organisms.This is important because jumping genes have come to be used as a mode of transferring modified gene segments into and between organisms in industrial laboratories.Now,they are seen as being involved in such phenomena as increasing antibiotic resistance in bacteria and fungi(see papers by Hanage et al 2009 and Graham 2009)and spread of introduced gene sequences from fungi to aphids(Moran and Jarvik 2010)and from plants to coffee pests(Ramanujan 2012). Recognizing the potential impacts,now China a leading GMO developer and user,has banned use of genetic engineering for their main food staple, rice. Most recently,a report this year from the European Food Safety Authority has discovered a viral sequence contained within commercial GMOs that carries a virus(see report by Podevin and du Jardin 2012). They found that 54 of 86 insertions of foreign DNA for cauliflower mosaic virus.This means that we are spreading a virus previously undetected by the corporations who use these sequences in the GMOs currently in use. This information is available freely on the web.As time goes by, more and more such information will become available. It should be considered seriously in your own debates concerning use of GMOs in our small island community.Thank you for your time and patience. ••■, ,4 ..►v. Comm. No. �J I' /it ---- Ref. To: 'JLil�'7 a1�L Dr.William .M.Steiner,CEO,PALMS Ref. Date ' '• A DIVISION OF STEINER-PACIFIC ENTERPRISES—SINCE 1969 Post Office Box 4565,Hilo,HI 96720 tel.808-294-0750 Science 9 November 2012: Vol. 338 no. 6108 pp. 758-767 DOI: 10.1126/science.338.6108.758 • ASSOCIATION AFFAIRS PRESIDENTIAL ADDRESS Transposable Elements, Epigenetics,and Genome Evolution . Nina V. Fedoroff Author Affiliations . Nina V. Fedoroff is Distinguished Professor of Biosciences at the King Abdullah University of Science and Technology, Saudi Arabia, and Evan Pugh Professor in the Huck Institutes of the Life Sciences, Pennsylvania State University. She served as the Science and Technology Adviser to the Secretary of State and to the Administrator of the U.S. Agency for International Development (USAID)from 2007 to 2010. She was president of AAAS from February 2011 to February 2012. This article is based on the Presidential Address she delivered at the AAAS annual meeting in Vancouver, BC,on 16 February 2012. . E-mail: nvfl @psu.edu View larger version: • In this page In a new window • Download PowerPoint Slide for Teaching Fig. 1. The C-value paradox. The range of haploid genome sizes is shown in kilobases for the groups of organisms listed on the left. [Adapted from an image by Steven M. Carr, Memorial University of Newfoundland] Transposable genetic elements(TEs)comprise a vast array of DNA sequences,all having the ability to move to new sites in genomes either directly by a cut-and-paste mechanism (transposons)or indirectly through an RNA intermediate(retrotransposons). First discovered in maize plants by the brilliant geneticist Barbara McClintock in the mid-I 940s,they were initially considered something of a genetic oddity(1, 2). Several decades later, TEs acquired the anthropomorphic labels of"selfish" and "parasitic" because of their replicative autonomy and potential for genetic disruption(3,4). However, TEs generally exist in eukaryotic genomes in a reversibly inactive, genetically undetectable form we now call "epigenetically silenced,"whose discovery can also be traced to McClintock's elegant genetic studies(5, 6). As the underlying biochemical mechanisms emerged from obscurity and epigenetics became popular toward the end of the 20th century, it was proposed that epigenetic silencing evolved to control the proliferation of TEs and their perceived destructive potential (5,6). Today,we know that TEs constitute more than half of the DNA in many higher eukaryotes. We know, too,that the fingerprints of TEs and transposition are everywhere in their genomes,from the coarsest features of genomic landscapes and how they change through real and evolutionary time to the finest details of gene structure and regulation. My purpose here is to challenge the current, somewhat pejorative, view of TEs as genomic parasites with the mounting evidence that TEs and transposition play a profoundly generative role in genome evolution. I contend that it is precisely the elaboration of epigenetic mechanisms from their prokaryotic origins as suppressors of genetic exchanges that underlies both the genome expansion and the proliferation of TEs characteristic of higher eukaryotes. This is the inverse of the prevailing view that epigenetic mechanisms evolved to control the disruptive potential of TEs. The evidence that TEs shape eukaryotic genomes is by now incontrovertible. My thesis,then, is that TEs and the transposases they encode underlie the evolvability of higher eukaryotes' massive, messy genomes. Although my examples in this essay are largely from plants,I believe that the inferences drawn apply to higher eukaryotes in general, among which plants tend toward exaggeration in genome size, TE abundance, and epigenetic complexity. Perhaps because they have no recourse to behavioral responses in coping with stressful environments,plants appear to have honed genetic and epigenetic strategies for adaptation to a much greater extent than animals. HOW TRANSPOSONS CAME TO BE CALLED "SELFISH" DNA View larger version: • In this page In a new window • Download PowerPoint Slide for Teaching Fig.2. Generation and elimination of duplications by unequal crossing over. Broken lines trace the recombination event. The invention of DNA sequencing techniques in the late 1970s and their subsequent mechanization led to an explosion of knowledge about the structure, gene content,and organization of genomes.The 1960s had seen the development of nucleic acid reassociation techniques whose application revealed the presence of much repetitive DNA in eukaryotic genomes(7, 8). As DNA sequencing became a reality, a good deal of discussion arose over the value of sequencing entire genomes, particularly that of humans(9-12), in view of the calculation that only a tiny fraction of the genome consisted of genes in the then-conventional sense of protein- and structural RNA–coding sequences and their associated regulatory sequences (13). A pair of papers published in Nature in 1980 solidified the idea that much of eukaryotic DNA, including transposons,was "junk"—a designation conferred a decade earlier by Ohno, who argued that our genomes were replete with nonfunctional DNA(3, 4, 14). The objective of the Nature papers was to get beyond the then still-prevalent view that every bit of an organism's DNA has a specific function crafted by selection. Thus,both papers promoted Dawkins' concept of"selfish DNA"—the notion that DNA capable of proliferating within a genome, as TEs do, may need no other explanation for its survival (15). Orgel and Crick asserted, "The spread of selfish DNA sequences within the genome can be compared to the spread of a not-too-harmful parasite within its host"(3). The selfish DNA concept was initially offered to explain the long-standing C-value paradox that organisms of similar evolutionary complexity differ vastly in their DNA content(16), and this it did. The C value, which is the DNA content per haploid genome,varies widely among closely related organisms of apparently comparable complexity (Fig. 1);this has for some time been attributed to the repetitive portion of the genome(17). Such variation is especially striking in angiosperms, whose highest and lowest C values differ by a factor of 2000(18, 19). The explanation of the C-value paradox does indeed reside largely in the profound differences among genomes in the abundance of TEs,primarily retrotransposons, even as gene numbers remain relatively constant. The Arabidopsis genome, for example,contains about 27,000 genes and 20 to 25 Mb of retrotransposons, whereas the maize genome contains about 40,000 genes and more than 1800 Mb of retrotransposon sequences(20-22). What the selfish DNA hypothesis does not attempt to explain,however, is how genomes can accumulate such vast amounts of repetitive sequences, given the ease of eliminating them by homologous recombination. THE SELFISH DNA LABEL STUCK View larger version: • In this page In a new window • Download PowerPoint Slide for Teaching y_ This Week in SCIENCE, Volume 324, Issue 5933 dated June 12 2009, is now available at: Recombining Resistance Homologous recombination is frequent in many bacteria, but few studies have addressed whether subpopulations within a species are more or less likely to undergo this process and whether it has consequences for their evolution. Taking a large data set from the pathogen Streptococcus pneumoniae, Hanage et al (p. 1454) discovered a group of strains characterized by an anomalous sequence of housekeeping genes. This sequence appeared to have been horizontally acquired from other pneumococci and related species and was associated with resistance to all classes of antibiotics for which data are available. Thus, hyper-recombination (in contrast to hypermutation) is important in the evolution and spread of antibiotic resistance and may play a role in determining the emergence of species clusters and the phenotypes associated with them. Science 12 June 2009:VoI. 324. no. 5933, pp. 1454 - Prey Tat 1457D01: 10.1126/science.1171908 REPORTS Hyper-Recombination, Diversity, and Antibiotic Resistance in Pneumococcus William Paul Hanage,'"" Christophe Fraser,' Jing Tang,2 Thomas Richard Connor,' Jukka Corander2 Streptococcus pneumoniae is a pathogen of global importance that frequently transfers genetic material between strains and on occasion across species boundaries. In an analysis of 1930 pneumococcal genotypes from six housekeeping genes and 94 genotypes from related species, we identified mosaic genotypes M representing admixture between populations and found that these were significantly associated with resistance to several classes of antibiotics.We hypothesize that these observations result from a history of hyper-recombination, which means that these strains are more likely to acquire both divergent genetic material and resistance determinants. This could have consequences for the reemergence of drug resistance after pneumococcal vaccination and also for our understanding of diversification and speciation in recombinogenic bacteria. 1 Department of Infectious Disease Epidemiology, Imperial College London, Norfolk Place, London W2 1PG, UK. 2 Department of Mathematics, Abo Akademi, FI-20500, Turku, Finland. To whom correspondence should be addressed. E-mail: w.hanage(@imperial.ac.uk Date: Sunday, January 3, 2010, 7:23 AM > from Science Daily <http://www.sciencedaily.com/releases/2009/12/091223125137.htm> > "Soil Studies Reveal Rise in Antibiotic Resistance" > ScienceDaily (Dec. 24, 2009) — Antibiotic resistance in > the natural environment is rising despite tighter controls > over our use of antibiotics in medicine and agriculture, > Newcastle University scientists have found. > Bacterial DNA extracted from soil samples collected between > 1940 and 2008 has revealed a rise in background levels of > antibiotic resistant genes. > Newcastle University's Professor David Graham, who led the > research, said the findings suggest an emerging threat to > public and environmental health in the future. > "Over the last few decades there has been growing concern > about increasing antibiotic resistance and the threat it > poses to our health, which is best evidenced by MRSA," > explained Professor Graham, who is based in the School of > Civil Engineering and Geosciences at Newcastle University. > "Despite increasingly stringent controls on our use of > antibiotics, the background level of antibiotic resistant > genes, which are markers for potential resistance, continues > to rise in soils." > "This increases the chances of a resistant gene in a > harmless bacteria being passed onto a disease-causing > pathogen, such as a MRSA, with obvious consequences." > Published online this week in the academic journal > Environmental Science and Technology, the report uses data > taken from five sites in the Netherlands. > The team found that 78 per cent of genes from four classes > of antibiotics showed increasing levels since 1940 — > despite continued efforts to reduce environmental levels. > Professor Graham said the next step would be to analyse > soil samples from other parts of the world, although he > expects to see similar results. > He adds: "The big question is that with more stringent > European regulations and greater emphasis on conservative > antibiotic use in agriculture and medicine, why are > antibiotic resistant gene levels still rising?" > "Whatever the cause, this rise suggests an ever increasing > risk of resistant genes being passed from environmental > organisms to organisms of greater health concern." > Professor Graham contends that more complementary studies > are desperately needed between environmental and public > health researchers to determine whether this increasing > 'pool' of resistance is actually contributing to harmful > bacteria, such as MRSA. > Story Source: > Adapted from materials provided by Newcastle University. > Journal Reference: Knapp et al. Evidence of Increasing > Antibiotic Resistance Gene Abundances in Archived Soils > since 1940. Environmental Science & Technology, 2009; > 091221172123091 DOI: 10.1021/es901221 x Science 30 April 2010:Vol. 328. no. 5978, pp. 624 - 627 Pre% science.1187113 Lateral Transfer of Genes from Fungi Underlies Carotenoid Production in Aphids Nancy A. Moroni,* and Tyler Jarvik2 Carotenoids are colored compounds produced by plants, fungi, and microorganisms and are required in the diet of most animals for oxidation control or light detection. Pea aphids display a red-green color polymorphism, which influences their susceptibility to natural enemies, and the carotenoid torulene occurs only in red individuals. Unexpectedly, we found that the aphid genome itself encodes multiple enzymes for carotenoid biosynthesis. Phylogenetic analyses show that these aphid genes are derived from fungal genes, which have been integrated into the genome and duplicated. Red individuals have a 30-kilobase region, encoding a single carotenoid desaturase that is absent from green individuals.A mutation causing an amino acid replacement in this desaturase results in loss of torulene and of red body color. Thus, aphids are animals that make their own carotenoids. 1 Department of Ecology and Evolutionary Biology, 1041 East Lowell Street, University of Arizona, Tucson, AZ 85721, USA. 2 Department of Chemistry and Biochemistry, University of Arizona, Tucson, AZ 85721, USA. To whom correspondence should be addressed. E-mail: nancy.moran@yale.edu Read the Full Text CORNELL CHRONICLE Feb.29,2012 Rare bacteria-to-animal gene transfer offers evolutionary advantage to coffee pest By Krishna Ramanujan A bacterial gene from a coffee pest's gut has evidently become a permanent part of the insect's genome,researchers have recently discovered. The finding points to a rare case of bacteria-to-animal horizontal gene transfer(HGT)— the nonsexual transmission of genetic material across species boundaries,reports a paper published Feb.27 in the Proceedings of the National Academy of Sciences.The transferred gene appears to give the coffee berry borer beetle(Hypothenemus hampei)an evolutionary advantage in digesting the carbohydrates found in coffee beans. The study may lead to ways of blocking the production of the enzyme in the coffee borer,which could potentially reduce the annual$500 million in losses the pest causes;the insect's damages also affect more than 20 million rural farming families.Native to Africa,the coffee borer has spread throughout the world,most recently to Hawaii. During the coffee borer's life cycle,female insects drill into coffee beans,where they lay eggs and eat the beans' storage carbohydrates,called galactomannans.The gene recently discovered in the coffee borer,called HhMAN1,encodes an enzyme(mannanase)that allows the insect to digest the galactomannan.This class of enzyme has never previously been reported in an insect but is commonly found in gut bacteria. Although there are many known cases of HGT between single-celled organisms(prokaryotes),such genetic transfer in animals is rare,and even rarer when there is an obvious evolutionary advantage. "The combination of intensive agricultural practices and a specific ecological niche may create selective pressures for horizontal gene transfer,"said Jocelyn Rose,a Cornell associate professor of plant biology and a senior author of the paper.Lead authors include Ricardo Acuna and Beatriz Padilla,both researchers at Cenicafe, Colombia's national coffee institute. No one knows precisely how such HGT occurs,but the researchers found that the HhMAN I gene sequence was flanked by transposons,or jumping genes,a category of genes known for their ability to move from place to place in DNA.The finding could represent a potential mechanism of gene transfer from gut bacteria to the insect,according to the researchers."At this point it's still more of a smoldering gun than a smoking gun"said Rose.The findings could lead the way,he said,to discovering other examples of HGT in agricultural settings. When sequencing genomes,computational biologists often put filters in place that remove bacterial sequences,which may mean scientists could be underestimating how frequently HGT occurs,Rose said. The work was funded by the Ministry of Agriculture and Rural Development of Colombia,National Federation of Coffee Growers of Colombia,Cornell's Atkinson Center for a Sustainable Future and the National Science Foundation. Regulators Discover a Hidden Viral Gene in Commercial GMO Crops January 21 , 2013 Biotechnology, Commentaries 15 Comments by Jonathan Latham and Allison Wilson Independent Science News How should a regulatory agency announce they have discovered something potentially very important about the safety of products they have been approving for over twenty years? In the course of analysis to identify potential allergens in GMO crops, the European Food Safety Authority (EFSA) has belatedly discovered that the most common genetic regulatory sequence in commercial GMOs also encodes a significant fragment of a viral gene (Podevin and du Jardin 2012). This finding has serious ramifications for crop biotechnology and its regulation, but possibly even greater ones for consumers and farmers. This is because there are clear indications that this viral gene (called Gene VI) might not be safe for human consumption. It also may disturb the normal functioning of crops, including their natural pest resistance. CAULIFLOWER MOSAIC VIRUS What Podevin and du Jardin discovered is that of the 86 different transgenic events (unique insertions of foreign DNA) commercialized to-date in the United States 54 contain portions of Gene VI within them. They include any with a widely used gene regulatory sequence called the CaMV 35S promoter (from the cauliflower mosaic virus; CaMV). Among the affected transgenic events are some of the most widely grown GMOs, including Roundup Ready soybeans (40-3-2) and MON810 maize. They include the controversial NK603 maize recently reported as causing tumors in rats (Seralini et al. 2012). The researchers themselves concluded that the presence of segments of Gene VI "might result in unintended phenotypic changes". They reached this conclusion because similar fragments of Gene VI have already been shown to be active on their own (e.g. De Tapia et al. 1993). In other words, the EFSA researchers were unable to rule out a hazard to public health or the environment. In general, viral genes expressed in plants raise both agronomic and human health concerns (reviewed in Latham and Wilson 2008). This is because many viral genes function to disable their host in order to facilitate pathogen invasion. Often, this is achieved by incapacitating specific anti-pathogen defenses. Incorporating such genes could clearly lead to undesirable and unexpected outcomes in agriculture. Furthermore, viruses that infect plants are often not that different from viruses that infect humans. For example, sometimes the genes of human and plant viruses are interchangeable, while on other occasions inserting plant viral fragments as transgenes has caused the genetically altered plant to become susceptible to an animal virus (Dasgupta et al. 2001). Thus, in various ways, inserting viral genes accidentally into crop plants and the food supply confers a significant potential for harm. The Choices for RegulatorsThe original discovery by Podevin and du Jardin (at EFSA) of Gene VI in commercial GMO crops must have presented regulators with sharply divergent procedural alternatives. They could 1) recall all CaMV Gene VI-containing crops (in Europe that would mean revoking importation and planting approvals) or, 2) undertake a retrospective risk assessment of the CaMV promoter and its Gene VI sequences and hope to give it a clean bill of health. It is easy to see the attraction for EFSA of option two. Recall would be a massive political and financial decision and would also be a huge embarrassment to the regulators themselves. It would leave very few GMO crops on the market and might even mean the end of crop biotechnology. Regulators, in principle at least, also have a third option to gauge the seriousness of any potential GMO hazard. GMO monitoring, which is required by EU regulations, ought to allow them to find out if deaths, illnesses, or crop failures have been reported by farmers or health officials and can be correlated with the Gene VI sequence. Unfortunately, this particular avenue of enquiry is a scientific dead end. Not one country has carried through on promises to officially and scientifically monitor any hazardous consequences of GMOs (1). Unsurprisingly, EFSA chose option two. However, their investigation resulted only in the vague and unreassuring conclusion that Gene VI "might result in unintended phenotypic changes" (Podevin and du Jardin 2012). This means literally, that changes of an unknown number, nature, or magnitude may (or may not) occur. It falls well short of the solid scientific reassurance of public safety needed to explain why EFSA has not ordered a recall. Can the presence of a fragment of virus DNA really be that significant? Below is an independent analysis of Gene VI and its known properties and their safety implications. This analysis clearly illustrates the regulators' dilemma. The Many Functions of Gene VIGene VI, like most plant viral genes, produces a protein that is multifunctional. It has four (so far) known roles in the viral infection cycle. The first is to participate in the assembly of virus particles. There is no current data to suggest this function has any implications for biosafety. The second known function is to suppress anti-pathogen defenses by inhibiting a general cellular system called RNA silencing (Haas et al. 2008). Thirdly, Gene VI has the highly unusual function of transactivating (described below) the long RNA (the 35S RNA) produced by CaMV (Park et al. 2001). Fourthly, unconnected to these other mechanisms, Gene VI has very recently been shown to make plants highly susceptible to a bacterial pathogen (Love et al. 2012). Gene VI does this by interfering with a common anti-pathogen defense mechanism possessed by plants. These latter three functions of Gene VI (and their risk implications) are explained further below: 1) Gene VI Is an Inhibitor of RNA SilencingRNA silencing is a mechanism for the control of gene expression at the level of RNA abundance (Bartel 2004). It is also an important antiviral defense mechanism in both plants and animals, and therefore most viruses have evolved genes (like Gene VI) that disable it (Dunoyer and Voinnet 2006). GENE VI (UPPER LEFT) PRECEDES THE START OF THE 35S RNA This attribute of Gene VI raises two obvious biosafety concerns: 1) Gene VI will lead to aberrant gene expression in GMO crop plants, with unknown consequences and, 2) Gene VI will interfere with the ability of plants to defend themselves against viral pathogens. There are numerous experiments showing that, in general, viral proteins that disable gene silencing enhance infection by a wide spectrum of viruses (Latham and Wilson 2008). 2) Gene VI Is a Unique Transactivator of Gene ExpressionMulticellular organisms make proteins by a mechanism in which only one protein is produced by each passage of a ribosome along a messenger RNA (mRNA). Once that protein is completed the ribosome dissociates from the mRNA. However, in a CaMV-infected plant cell, or as a transgene, Gene VI intervenes in this process and directs the ribosome to get back on an mRNA (reinitiate) and produce the next protein in line on the mRNA, if there is one. This property of Gene VI enables Cauliflower Mosaic Virus to produce multiple proteins from a single long RNA (the 35S RNA). Importantly, this function of Gene VI (which is called transactivation) is not limited to the 35S RNA. Gene VI seems able to transactivate any cellular mRNA (Futterer and Hohn 1991 ; Ryabova et al. 2002). There are likely to be thousands of mRNA molecules having a short or long protein coding sequence following the primary one. These secondary coding sequences could be expressed in cells where Gene VI is expressed. The result will presumably be production of numerous random proteins within cells. The biosafety implications of this are difficult to assess. These proteins could be allergens, plant or human toxins, or they could be harmless. Moreover, the answer will differ for each commercial crop species into which Gene VI has been inserted. 3) Gene VI Interferes with Host DefensesA very recent finding, not known by Podevin and du Jardin, is that Gene VI has a second mechanism by which it interferes with plant anti-pathogen defenses (Love et al. 2012). It is too early to be sure about the mechanistic details, but the result is to make plants carrying Gene VI more susceptible to certain pathogens, and less susceptible to others. Obviously, this could impact farmers, however the discovery of an entirely new function for gene VI while EFSA's paper was in press, also makes clear that a full appraisal of all the likely effects of Gene VI is not currently achievable. Is There a Direct Human Toxicity Issue?When Gene VI is intentionally expressed in transgenic plants, it causes them to become chiorotic (yellow), to have growth deformities, and to have reduced fertility in a dose-dependent manner (Ziljstra et at 1996). Plants expressing Gene VI also show gene expression abnormalities. These results indicate that, not unexpectedly given its known functions, the protein produced by Gene VI is functioning as a toxin and is harmful to plants (Takahashi et al 1989). Since the known targets of Gene VI activity (ribosomes and gene silencing) are also found in human cells, a reasonable concern is that the protein produced by Gene VI might be a human toxin. This is a question that can only be answered by future experiments. Is Gene VI Protein Produced in GMO Crops?Given that expression of Gene VI is likely to cause harm, a crucial issue is whether the actual inserted transgene sequences found in commercial GMO crops will produce any functional protein from the fragment of Gene VI present within the CaMV sequence. There are two aspects to this question. One is the length of Gene VI accidentally introduced by developers. This appears to vary but most of the 54 approved transgenes contain the same 528 base pairs of the CaMV 35S promoter sequence. This corresponds to approximately the final third of Gene VI. Deleted fragments of Gene VI are active when expressed in plant cells and functions of Gene VI are believed to reside in this final third. Therefore, there is clear potential for unintended effects if this fragment is expressed (e.g. De Tapia et al. 1993; Ryabova et al. 2002; Kobayashi and Hohn 2003). The second aspect of this question is what quantity of Gene VI could be produced in GMO crops? Once again, this can ultimately only be resolved by direct quantitative experiments. Nevertheless, we can theorize that the amount of Gene VI produced will be specific to each independent insertion event. This is because significant Gene VI expression probably would require specific sequences (such as the presence of a gene promoter and an ATG [a protein start codon]) to precede it and so is likely to be heavily dependent on variables such as the details of the inserted transgenic DNA and where in the plant genome the transgene inserted. Commercial transgenic crop varieties can also contain superfluous copies of the transgene, including those that are incomplete or rearranged (Wilson et al 2006). These could be important additional sources of Gene VI protein. The decision of regulators to allow such multiple and complex insertion events was always highly questionable, but the realization that the CaMV 35S promoter contains Gene VI sequences provides yet another reason to believe that complex insertion events increase the likelihood of a biosafety problem. Even direct quantitative measurements of Gene VI protein in individual crop authorizations would not fully resolve the scientific questions, however. No-one knows, for example, what quantity, location or timing of protein production would be of significance for risk assessment, and so answers necessary to perform science-based risk assessment are unlikely to emerge soon. Big Lessons for Biotechnologylt is perhaps the most basic assumption in all of risk assessment that the developer of a new product provides regulators with accurate information about what is being assessed. Perhaps the next most basic assumption is that regulators independently verify this information. We now know, however, that for over twenty years neither of those simple expectations have been met. Major public universities, biotech multinationals, and government regulators everywhere, seemingly did not appreciate the relatively simple possibility that the DNA constructs they were responsible for encoded a viral gene. This lapse occurred despite the fact that Gene VI was not truly hidden; the relevant information on the existence of Gene VI has been freely available in the scientific literature since well before the first biotech approval (Franck et al 1980). We ourselves have offered specific warnings that viral sequences could contain unsuspected genes (Latham and Wilson 2008). The inability of risk assessment processes to incorporate longstanding and repeated scientific findings is every bit as worrysome as the failure to intellectually anticipate the possibility of overlapping genes when manipulating viral sequences. This sense of a generic failure is reinforced by the fact that this is not an isolated event. There exist other examples of commercially approved viral sequences having overlapping genes that were never subjected to risk assessment. These include numerous commercial GMOs containing promoter regions of the closely related virus figwort mosaic virus (FMV) which were not considered by Podevin and du Jardin. Inspection of commercial sequence data shows that the commonly used FMV promoter overlaps its own Gene VI (Richins et al 1987). A third example is the virus-resistant potato NewLeaf Plus (RBMT-22-82). This transgene contains approximately 90% of the PO gene of potato leaf roll virus. The known function of this gene, whose existence was discovered only after US approval, is to inhibit the anti-pathogen defenses of its host (Pfeffer et al 2002). Fortunately, this potato variety was never actively marketed. A further key point relates to the biotech industry and their campaign to secure public approval and a permissive regulatory environment. This has led them to repeatedly claim, firstly, that GMO technology is precise and predictable; and secondly, that their own competence and self-interest would prevent them from ever bringing potentially harmful products to the market; and thirdly, to assert that only well studied and fully understood transgenes are commercialized. It is hard to imagine a finding more damaging to these claims than the revelations surrounding Gene VI. Biotechnology, it is often forgotten, is not just a technology. It is an experiment in the proposition that human institutions can perform adequate risk assessments on novel living organisms. Rather than treat that question as primarily a daunting scientific one, we should for now consider that the primary obstacle will be overcoming the much more mundane trap of human complacency and incompetence. We are not there yet, and therefore this incident will serve to reinforce the demands for GMO labeling in places where it is absent. What Regulators Should Do NowThis summary of the scientific risk issues shows that a segment of a poorly characterized viral gene never subjected to any risk assessment (until now) was allowed onto the market. This gene is currently present in commercial crops and growing on a large scale. It is also widespread in the food supply. Even now that EFSA's own researchers have belatedly considered the risk issues, no one can say whether the public has been harmed, though harm appears a clear scientific possibility. Considered from the perspective of professional and scientific risk assessment, this situation represents a complete and catastrophic system failure. But the saga of Gene VI is not yet over. There is no certainty that further scientific analysis will resolve the remaining uncertainties, or provide reassurance. Future research may in fact increase the level of concern or uncertainty, and this is a possibility that regulators should weigh heavily in their deliberations. To return to the original choices before EFSA, these were either to recall all CaMV 35S promoter-containing GMOs, or to perform a retrospective risk assessment. This retrospective risk assessment has now been carried out and the data clearly indicate a potential for significant harm. The only course of action consistent with protecting the public and respecting the science is for EFSA, and other jurisdictions, to order a total recall. This recall should also include GMOs containing the FMV promoter and its own overlapping Gene VI. Footnotes1) EFSA regulators might now be regretting their failure to implement meaningful GMO monitoring. It would be a good question for European politicians to ask EFSA and for the board of EFSA to ask the GMO panel, whose job it is to implement monitoring. ReferencesBartel P (2004) MicroRNAs: Genomics, Biogenesis, Mechanism, and Function. Cell: 116, 281-297.Dasgupta R , Garcia BH, Goodman RM (2001) Systemic spread of an RNA insect virus in plants expressing plant viral movement protein genes. Proc. Natl. Acad. Sci. USA 98: 4910-4915. De Tapia M, Himmelbach A, and Hohn T (1993) Molecular dissection of the cauliflower mosaic virus translation transactivator. EMBO J 12: 3305-14. Dunoyer P, and 0 Voinnet (2006) The complex interplay between plant viruses and host RNA-silencing pathways. Curr Opinion in Plant Biology 8: 415-423. Franck A, H Guilley, G Jonard, K Richards and L Hirth (1980) Nucleotide sequence of cauliflower mosaic virus DNA. Cell 2: 285-294.Futterer J, and T Hohn (1991) Translation of a polycistronic mRNA in presence of the cauliflower mosaic virus transactivator protein. EMBO J. 10: 3887-3896. Haas G, Azevedo J, Moissiard G, Geldreich A, Himber C, Bureau M, et al. (2008) Nuclear import of CaMV P6 is required for infection and suppression of the RNA silencing factor DRB4. EMBO J 27: 2102-12. Kobayashi K, and T Hohn (2003) Dissection of Cauliflower Mosaic Virus TransactivatorNiroplasmin Reveals Distinct Essential Functions in Basic Virus Replication. J. Virol. 77: 8577-8583. Latham JR, and AK Wilson (2008) Transcomplementation and Synergism in Plants: Implications for Viral Transgenes? Molecular Plant Pathology 9: 85- 103. Park H-S, Himmelbach A, Browning KS, Hohn T, and Ryabova LA (2001). A plant viral "reinitiation" factor interacts with the host translational machinery. Cell 106: 723-733. Pfeffer S, P Dunoyer, F Heim, KE Richards, G Jonard, V Ziegler-Graff (2002) PO of Beet Western Yellows Virus Is a Suppressor of Posttranscriptional Gene Silencing. J. Virol. 76: 6815-6824. Podevin N and du Jardin P (2012) Possible consequences of the overlap between the CaMV 35S promoter regions in plant transformation vectors used and the viral gene VI in transgenic plants. GM Crops and Food 3: 1-5. Love AJ , C Geri, J Laird, C Carr, BW Yun, GJ Loake et al (2012) Cauliflower mosaic virus Protein P6 Inhibits Signaling Responses to Salicylic Acid and Regulates Innate Immunity. PLoS One. 7(10): e47535. Richins R, H Scholthof, RJ Shepherd (1987) Sequence of figwort mosaic virus DNA (caulimovirus group). NAR 15: 8451-8466. Ryabova LA , Pooggin, MH and Hohn, T (2002) Viral strategies of translation initiation: Ribosomal shunt and reinitiation. Progress in Nucleic Acid Research and Molecular Biology 72: 1-39. Seralini, G-E., E. Clair, R. Mesnage, S. Gress, N. Defarge, M. Malatesta, D. Hennequin, J. Spiroux de VendOmois. 2012. Long term toxicity of a Roundup herbicide and a Roundup-tolerant genetically modified maize. Food Chem. Toxicol. Takahashi H, K Shimamoto, Y Ehara (1989) Cauliflower mosaic virus gene VI causes growth suppression, development of necrotic spots and expression of defence-related genes in transgenic tobacco plants. Molecular and General Genetics 216:188-194. Wilson AK, JR Latham and RA Steinbrecher (2006) Transformation-induced mutations in transgenic plants: Analysis and biosafety implications. Biotechnology and Genetic Engineering Reviews 23: 209-234. Zijlstra C, Scharer-Hernandez N, Gal S, Hohn T. Arabidopsis thaliana expressing the cauliflower mosaic virus ORF VI transgene has a late flowering phenotype. Virus Genes 1996; 13:5-17. New study finds antibiotic resistance from GMOs in microbes in rivers Monday, 07 January 2013 19:11 COMMENT by UC Berkeley microbiologist Dr Ignacio Chapela: Paper demonstrating the escape and establishment of transgenic DNA from GMOs in all rivers tested in China. Salient points: 1.A novel method was used to select and isolate those bacteria from the river water which contained DNA suspected to derive from GMOs. These were then carefully studied to demonstrate that the DNA could not have come from other sources (e.g. mutation/natural selection). 2. Although the numbers varied, and absolute amounts of the transformed bacteria could not be measured, every river sampled was shown to contain these organisms. 3. The exact origin of the transgenic DNA incorporated into native bacterial populations was not determined. It could come from intentional releases (such as agricultural fields) or from unintentional escapes from contained situations (labs, industrial facilities). 4. These bacteria all have in common the fact that they acquired antibiotic resistance markers from transgenic origin (the markers were used to select the bacteria in the first place). Antibiotic resistance in free- living bacteria is not a good thing for those who may one day want to use antibiotics to fend-off infections (e.g. patients in hospitals). 5. Nevertheless, the antibiotic resistance is not at all the most important point of this paper (even when the authors themselves seem to think it is). Looking for antibiotic resistance was the easiest feasible way to do this work and it also has the obvious medical implications, but this is only a fraction of the many other sequences of transgenic DNA which must be expected out there in the environment, from all kinds of origins, with all kinds of possible functions. This paper is the equivalent of the proverbial sighting of the iceberg's tip. A polaroid photo of a small part of what must be a very large and relevant phenomenon. 6. The question which beggars belief is: why is it that nobody in the "West" has been able to follow up on such leads, or even suggest doing this kind of work? One thinks of the work begun at NYU on soils, which attracted so much negative campaigning, and nary a recognition. ------ Environ Sci Technol. 2012 Dec 18;46(24):13448-54. doi: 10.1021/es302760s. Epub 2012 Dec 6. http://www.ncbi.nlm.nih.gov/pubmed/23215020 A Survey of Drug Resistance bla Genes Originating from Synthetic Plasmid Vectors in Six Chinese Rivers. Chen J, Jin M, Qiu ZG, Guo C, Chen ZL, Shen ZQ, Wang XW, Li JW. College of Life Sciences, Sichuan University , Chengdu, Sichuan Province 610064, People's Republic of China. Abstract Antibiotic resistance poses a significant challenge to human health and its rate continues to rise globally. While antibiotic-selectable synthetic plasmid vectors have proved invaluable tools of genetic engineering, this class of artificial recombinant DNA M sequences with high expression of antibiotic resistance genes presents an unknown risk beyond the laboratory setting. Contamination of environmental microbes with synthetic plasmid vector-sourced antibiotic resistance genes may represent a yet unrecognized source of antibiotic resistance. In this study, PCR and real-time quantitative PCR were used to investigate the synthetic plasmid vector-originated ampicillin resistance gene, 13-lactam antibiotic (bla), in microbes from six Chinese rivers with significant human interactions. Various levels of bla were detected in all six rivers, with the highest levels in the Pearl and Haihe rivers. To validate the bla pollution, environmental plasmids in the river samples were captured by the E. coli transformants from the community plasmid metagenome. The resultant plasmid library of 205 ampicillin-resistant E. coli (transformants) showed a bla-positive rate of 27.3% by PCR. Sequencing results confirmed the synthetic plasmid vector sources. In addition, results of the Kirby-Bauer disc-diffusion test reinforced the ampicillin- resistant functions of the environmental plasmids. The resistance spectrum of transformants from the Pearl and Haihe rivers, in particular, had expanded to the third- and fourth-generation of cephalosporin drugs, while that of other transformants mainly involved first- and second-generation cephalosporins. This study not only reveals environmental contamination of synthetic plasmid vector-sourced bla drug resistance genes in Chinese rivers, but also suggests that synthetic plasm id vectors may represent a source of antibiotic resistance in humans. PMID:23215020 [PubMed - in process] 4 /.. .. SCENTER F O R FOOD SAFETY Contacts: Kevin Golden, 415-826-2770 Joseph Mendelson, (202) 547-9359 FEDERAL COURT UPHOLDS BAN ON GENETICALLY-ENGINEERED ALFALFA Appeals Court Rules Planting of Biotech Crop Can Cause Irreversible Harm to Conventional Varieties of Crops, Farmers, and the Environment Washington,DC- September 2, 2008 — In a decision handed down today, the United States Court of Appeals for the Ninth Circuit has upheld a nationwide ban on the planting of genetically-engineered (GE) Roundup Ready alfalfa pending a full Environmental Impact Statement(EIS). The Court determined that the planting of genetically modified alfalfa can result in potentially irreversible harm to organic and conventional varieties of crops,damage to the environment, and economic harm to farmers. Although the suit was brought against United States Department of Agriculture(USDA); Forage Genetics and Monsanto Company entered into the suit as Defendant-Intervenors. In her opinion,Circuit Judge Mary M. Schroeder held that"Monsanto and Forage Genetics contend that the District Court disregarded their financial losses, but the district court considered those economic losses and simply concluded that the harm to growers and consumers who wanted non-genetically engineered alfalfa outweighed the financial hardships to Monsanto and Forage Genetics and their growers." "This ruling affirms a major victory for consumers, ranchers, organic farmers, and most conventional farmers across the country," said Andrew Kimbrell, Executive Director of the Center for Food Safety. "Roundup Ready Alfalfa represents a very real threat to farmers' livelihoods and the environment; the judge rightly dismissed Monsanto's claims that their bottom line should come before the rights of the public and America's farmers. This ruling is a turning point in the regulation of biotech crops in this country." Today's decision upholds District Court Judge Charles Breyer's earlier ruling of May 2007, in which he found that the USDA failed to address concerns that Roundup Ready alfalfa will contaminate conventional and organic alfalfa. Judge Schroeder's decision affirms that USDA violated national environmental laws by approving GE alfalfa without a full Environmental Impact Statement. It also affirms that USDA failed to address the problem of Roundup-resistant "superweeds" that could follow commercial planting of GE alfalfa. The Center for Food Safety represented itself and the following co-plaintiffs in the suit: Western Organization of Resource Councils, National Family Farm Coalition, Sierra Club, Beyond Pesticides, Cornucopia Institute, Dakota Resource Council,Trask Family Seeds, and Geertson Seed Farms. For more information, please visit www.centerforfoodsafety.org • China Rejects Genetically Modified Rice in Opposition to GMO Crops Andre Evans NaturalSociety February 2, 2012 As awareness of genetically modified food and the dangers associated increase, actions are being taken to eliminate the issue entirely. With countries destroying their GMO crops, scientists showing the facts, and individuals voicing their discontent on the matter, there is a large and growing contingent across countries that are taking action to preserve the integrity of our food supply. China is the latest nation to take a stand against GMO crops. China recently suspended the distribution of genetically modified rice within its commercial food supplies. Rice, being perhaps the most common and prominently used food within Chinese diet makes its safety and reliability very significant. Acting as a staple food for over 1.3 billion Chinese people, any compromise to the integrity of their food supply should be accordingly assessed and acted against based upon the interests of the population. This action helps to preserve the sovereignty and safety of their food supply, something that largely must still be addressed in our own country. In the United States GMO foods are not only accepted by loose regulation standards, but are not even labeled appropriately. The dangers of GMO foods are so great that they are still not — and may never be — fully assessable. With a large number of documented negative effects on its consumers, GMO foods are a highly dangerous threat to individual health alone. If this weren't enough, genetically engineered crops actually threaten the integrity of the food supply altogether by devastating the environment with powerful superweeds and mutant bugs. The nature of genetic engineering is so volatile that unless properly handled, the integrity of the entire world food supply could be ruined. As such, any form of action that a government takes to prevent or eliminate GMOs from the equation is significant. Reassessing the nature and scale of the threat in light of new evidence should compel even more action in even more locations around the world. > Subject: Scientists warn EPA on Monsanto corn rootworm > http://www.reuters.com/article/2012/03/09/us-monsanto-corn- idUS BRE82815Z20120309 > Scientists warn EPA on Monsanto corn rootworm > • Analysis: EU farmers lose out as consumers oppose CMOs<http://www.reuters.com/article/2012/02/17/us-qmo-europe- idUSTRE81 G1A720120217> > Fri, Feb 17 2012 > • Monsanto guilty of chemical poisoning in France<http://www.reuters.com/article/2012/02/13/us-france-pesticides- monsanto-idUSTRE81 COVQ20120213> > Mon, Feb 13 2012 > • Origin expects to grow China's first GMO corn in 2013<http://www.reuters.com/article/2012/02/10/u s-china-oriq i n- idUSTRE8190JV20120210> > Fri, Feb 10 2012 > By Carey Gillam<http://blogs.reuters.com/search/journalist.php?edition=us&n=carey.gillam &> > Fri Mar 9, 2012 3:46pm EST > (Reuters) - A group of plant scientists is warning federal regulators that action is needed to mitigate a growing problem with biotech corn that is losing its resistance to plant-damaging pests. > > The stakes are high - corn production is critical for food, animal feed and ethanol production, and farmers have increasingly been relying on corn that has been genetically modified to be toxic to corn rootworm pests. > "This is not something that is a surprise... but it is something that needs to be addressed," said Joseph Spencer, a corn entomologist with the Illinois Natural History Survey, part of the University of Illinois. > Spencer is one of 22 academic corn experts who sent a letter dated March 5 to the Environmental Protection Agency telling regulators they are worried about long-term corn production prospects because of the failure of the genetic modifications in corn aimed at protection from rootworm. > Monsanto introduced its corn rootworm protected products, which contain a protein referred to as "Cry3Bb1," in 2003 and they have proved popular with farmers in key growing areas around the country. Biotech corn sales are a key • GMO feeding Impacts By Dr. Mercola The first-ever lifetime feeding study 1 evaluating the health risks of genetically engineered foods was published online on September 19, and the results are troubling,to say the least. This new study joins a list of over 30 other animal studies showing toxic or allergenic problems with genetically engineered foods. The study,published in the peer-reviewed journal Food and Chemical Toxicology,found that rats fed a type of genetically engineered corn that is prevalent in the US food supply for two years developed massive mammary tumors,kidney and liver damage, and other serious health problems. The research was considered so "hot"that the work was done under strict secrecy. According to a French article in Le Nouvel Observateur,2 the researchers used encrypted emails,phone conversations were banned, and they even launched a decoy study to prevent sabotage! According to the authors: "The health effects of a Roundup-tolerant genetically modified maize (from 11% in the diet), cultivated with or without Roundup, and Roundup alone (from 0.1ppb in water), were studied 2 years in rats. [Editors note:this level of Roundup is permitted in drinking water and GE crops in the US] In females, all treated groups died 2-3 times more than controls, and more rapidly. This difference was visible in 3 male groups fed GMOs. All results were hormone and sex dependent, and the pathological profiles were comparable. Females developed large mammary tumors almost always more often than and before controls, the pituitary was the second most disabled organ; the sex hormonal balance was modified by GMO and Roundup treatments. In treated males, liver congestions and necrosis were 2.5-5.5 times higher... Marked and severe kidney nephropathies were also generally 1.3-2.3 greater. Males presented 4 times more large palpable tumors than controls, which occurred up to 600 days earlier. Biochemistry data confirmed very significant kidney chronic deficiencies;for all treatments and both sexes, 76%of the altered parameters were kidney related. These results can be explained by the non linear endocrine-disrupting effects of Roundup, but also by the overexpression of the transgene in the GMO and its metabolic consequences." Folks, if this doesn't get your attention,nothing will. Does 10 percent or more of your diet consist of genetically engineered(GE)ingredients?At present,you can't know for sure,since GE foods are not labeled in the US. But chances are, if you eat processed foods,your diet is chock full of genetically engineered ingredients you didn't even know about. The study in question includes photos and graphs. I highly recommend taking the time to actually read through this remarkable study,3 and look at the documented evidence. They really are not exaggerating when they say it caused massive tumors... They are huge! Some of the tumors weighed in at 25 percent of the rat's total body weight. This is the most current and best evidence to date of the toxic effects of GE foods. Why Aren't Americans Dropping Like Flies? Rats only live a few years. Humans live around 80 years,so we will notice these effects in animals long before we see them in humans. The gigantic human lab experiment is only about 10 years old, so we are likely decades away from tabulating the human casualties. This is some of the strongest evidence to date that we need to exercise the precautionary principle ASAP and avoid these foods. Naturally,the study is already under heavy fire. According to Monsanto spokesman Thomas Helscher:4 "Numerous peer-reviewed scientific studies performed on biotech crops to date, including more than a hundred feeding studies, have continuously confirmed their safety, as reflected in the respective safety assessments by regulatory authorities around the world." However, it's critical to understand that the longest feeding study was a mere 90 days long—a far cry from two years! In the featured study,the true onslaught of diseases really set in during the 13th month of the experiment, although tumors and severe liver and kidney damage did emerge as early as four months in males, and seven months for females. Is it any wonder then that feeding studies lasting just a few weeks or even three months have failed to corroborate these horrific findings?Reuters quotes Mark Tester,a research professor at the Australian Centre for Plant Functional Genomics at the University of Adelaide as saying:5 "If the effects are as big as purported, and if the work really is relevant to humans, why aren't the North Americans dropping like flies? GM has been in the food chain for over a decade over there —and longevity continues to increase inexorably." Although there are clearly many variables that contribute to cancer, GE foods are a new candidate as they have been in our food supply for over a decade. Interestingly,cancer was just declared as having overtaken heart disease as the number one killer among American Hispanics,6 and according to 2009 CDC statistics it's now also the leading killer in 18 states. I believe it is crucial that we implement the precautionary principle as rapidly as possible as this study confirms it is difficult to predict precisely what GE foods might do to the youths of today, as many are eating a fair amount of GE ingredients practically from day one. (Yes,some infant formulas actually contain GE ingredients!) What will their health be 10 or 20 years from now? Most adults simply haven't been eating GE foods long enough to tell what the real ramifications B (.t1 any 04 American Journal of Botany 99(4):700-707.2012. v EVIDENCE OF REDUCED ARBUSCULAR MYCORRHIZAL FUNGAL COLONIZATION IN MULTIPLE LINES OF BT MAIZE1 TANYA E.CHEEKE2,TODD N. ROSENSTIEL,AND MITCHELL B.CRUZAN Portland State University,Department of Biology,P.O.Box 751,Portland,Oregon 97207 USA • Premise of the study:Insect-resistant Bacillus thuringiensis(Bt)maize is widely cultivated,yet few studies have examined the interaction of symbiotic arbuscular mycorrhizal fungi(AMF)with different lines of Bt maize.As obligate symbionts,AMF may be sensitive to genetic changes within a plant host.Previous evaluations of the impact of Bt crops on AMF have been inconsistent,and because most studies were conducted under disparate experimental conditions,the results are difficult to compare. • Methods:We evaluate AMF colonization in nine Bt maize lines,differing in number and type of engineered trait,and five corresponding near-isogenic parental(P)base hybrids in greenhouse microcosms.Plants were grown in 50%local agricultural soil with low levels of fertilization,and AMF colonization was evaluated at 60 and 100 d.Nontarget effects of Bt cultivation on AMF colonization were tested in a subsequently planted crop,Glycine max,which was seeded into soil that had been pre- conditioned for 60 d with Bt or P maize. • Key results:We found that Bt maize had lower levels of AMF colonization in their roots than did the non-Bt parental lines. However,reductions in AMF colonization were not related to the expression of a particular Br protein.There was no difference in AMF colonization in G.max grown in the Br-or P-preconditioned soil. • Conclusions:These findings are the first demonstration of a reduction in AMF colonization in multiple Bt maize lines grown under the same experimental conditions and contribute to the growing body of knowledge examining the unanticipated effects of Bt crop cultivation on nontarget soil organisms. Key words: arbuscular mycorrhizal fungi;Bacillus thuringiensis;Cry lAb;Cry34/35Abl;Cry3Bb1;Cry1F;Glycine max; soybean;transgenic;Zea mays. Genetically modified(GM)crops,engineered to express her- To date,more than 60 different Bt crystal proteins(called Cry bicide-tolerance, insecticidal properties, or a combination of proteins)that exhibit a high degree of specificity toward certain traits,are the most rapidly adopted agricultural biotechnology insect pests have been identified (reviewed in Schnepf et al., in recent history (James, 2010). Since their commercial intro- 1998;Federici,2002;Stotzky,2002;Lee et al.,2003;Icoz and duction in 1996, the global adoption of GM crop technology Stotzky,2008b;Sanchis,2011).Bt crops that provide resistance has increased ca.87-fold,up from 1.7 million hectares in 1996 to multiple agricultural pests,as well as confer herbicide-tolerance, to 148 million hectares in 2010(James,2010). Insect-resistant have contributed to the popularity of GM crops among farmers maize (Zea mays L.), one of the most widely cultivated GM worldwide(EPA,2011). In 2010, 86% of the maize grown in crops,is engineered to express insecticidal toxins derived from the USA(USDA,2010)and 26%of the global biotech hectarage the spore-forming soil bacterium Bacillus thuringiensis (Bt). was cultivated in maize genetically modified to express one or more engineered traits (James, 2010). This rapid and wide- spread adoption of GM crops has led to a dramatic shift in the 'Manuscript received 2 November 2011;revision accepted 10 February 2011. agricultural landscape over the last 15 years and has raised The authors thank members of the Cruzan laboratory for research questions about the impact of insect-resistant Bt crops on non- assistance and C.A.Miles and B.Wolfley for the field soil used in this target organisms in the soil environment. experiment.Maize seed was provided by Syngenta Seeds,Monsanto Co., Arbuscular mycorrhizal fungi(AMF)are obligate plant sym- and an additional seed industry representative. Glycine max seed was bionts that have been shown to improve plant nutrient acquisi- provided by Territorial Seed Co., Cottage Grove, OR,USA. This work tion,especially in low nutrient soil environments(e.g.,Galvez benefited from valuable feedback provided by members of the Cruzan and 2001;Gosling et al.,2006;Lekberg et al., 2008; Sheng Bever-Schultz laboratory groups and two anonymous reviewers.Feedback on an earlier version of this manuscript was provided by Monsanto Co.and et al., 2008).These symbiotic fungi are ubiquitous in soil and an additional seed company representative.Funding for this research was are found in both natural and agroecosystems(Smith and Read, provided by grants from the Charles A. and Anne Morrow Lindbergh 2008).Because AMF rely on a plant host for nutrition and re- Foundation,the U.S.Environmental Protection Agency(EPA)Science to production,they may be sensitive to changes in the physiology Achieve Results (STAR) Graduate Fellowship Program, Sigma Delta of the host plant,to biochemical changes associated with the Bt Epsilon Graduate Women in Science, the National Science Foundation modification,or to alterations in root exudates released into the (DEB-1011525), Sigma Xi, Botanical Society of America, and a PSU rhizosphere.Although Bt proteins are expressed in the roots of Miller Grant for Sustainability.The funding agencies have not officially most Bt maize lines(Saxena and Stotzky,2000;Saxena et al., endorsed this publication,and the views expressed herein may not reflect 2002;reviewed by Icoz and Stotzky,2008a,b;EPA,2011),the the views of the funding agencies. 2Author for correspondence(e-mail: cheeket @pdx.edu),phone: (503) evidence that Cry proteins have a direct effect on AMF is 725-3801,fax:(503)725-3888 equivocal. For example, lower AMF colonization levels have been reported in Bt maize lines Bt 11 (Castaldini et al., 2005; doi:10.3732/ajb.1100529 Cheeke et al.,2011)and Bt 176(Turrini et al.,2004;Castaldini American Journal of Botany 99(4):700-707,2012;http://www.amjbotorg,/0 2012 Botanical Society of America 700 'Escaped' Genetically Engineered Canola Growing Outside of Established Cultivation Regions Across North Dakota Oct. 6, 2011 — Large, persistent populations of genetically engineered canola 1 have been found outside of cultivation in North Dakota. As genetically engineered crops become increasingly prevalent in the United States, concerns remain about potential ecological side effects. A study published by the online journal PLoS ONE reports that genetically engineered canola endowed with herbicide resistance have been found growing outside of established cultivation regions along roadsides across North Dakota. These "escaped" plants were found state-wide and accounted for 45% of the total roadside plants sampled. Furthermore, populations were found to persist from year to year and reach thousands of individuals. The authors also found that the escaped plants could hybridize with each other to create novel combinations of transgenic traits. The authors argue that their result, more than 10 years after the initial release of genetically engineered canola, "raises questions of whether adequate oversight and monitoring protocols are in place in the U.S. to track the environmental impact of biotech products." However, they also note that biotechnology can provide important tools to feed the rapidly growing population. "We must safely engage all tools available to us to advance food, fuel and fiber alternatives as modern agriculture rises to the challenges of the next decade," they conclude. "More than half of the Earth's terrestrial landscape is managed in cultivated crops or forage species," says lead researcher Cynthia Sagers, "yet we have little understanding of how domesticated plants influence their wild relatives. This study is a first step in addressing these questions by documenting that domesticated species have a life outside of cultivated fields." GM soy linked to health damage in pigs - a Danish Dossier Friday, 27 April 2012 21:10 Introduction A Danish farming newspaper has caused quite a stir by devoting a sizeable part of its 13 April edition to the discoveries by pig farmer lb Borup Pedersen that GM soy has a damaging effect both on his animals and on his farming profitability. On the front page of the paper there was a lead story under the headline "Pig farmer reaps gains from GMO-free soy". On a sidebar the paper referred to Mr Pedersen's contention that DDT and Thalidomide were minor problems when set alongside GMOs and Glyphosate. In an Editorial Comment on page 2, the paper argued that it would be grossly irresponsible for the authorities to ignore or ridicule the discoveries made by the farmer in his pig farming operations, and it congratulated the authorities for commissioning a new study designed to determine whether stomach lesions and other effects might be associated with GM soy; in the study 100 animals will be fed with non-GM soy and 100 with GM soy in their diets. On pages 6 and 7 of the paper there was a big article written by Anne Wolfenberg, who is a very experienced journalist who knows the Danish pig farming industry well. This article was leaked in draft form, translated into English and widely circulated, appearing on various web sites. GM-Free Cymru helped with that translation, in the belief that this was the final published version and that the farmer, the writer and the newspaper would be happy to see it circulated to an English-speaking readership. Full acknowledgement and citation were made. However, we did not realise that there were one or two small errors in the draft which were corrected in the final printed version; and partly on that basis we received a complaint from the author. We apologised for the misunderstanding, and the article was immediately removed from the GM Watch web site. We also asked an American web site which had used the article to take it down, in line with the journalist's wishes. This was also done. In deference to the concerns of Anne Wolfenberg, we are not including any translation of her article here. Instead, we have spoken to Mr Pedersen, and he has kindly given us permission to use content from his Powerpoint presentation, to use direct statements made by him, and to use his photographs. On that basis we have assembled the first part of this dossier. In the second part of the dossier, we examine the key points arising from the coverage of this issue in the Danish farming newspaper Effektivt Landbrug. We congratulate the editor and the journalists involved on a very effective and well-researched piece of investigative journalism, and for having the courage to publish it in spite of the anger it was bound to provoke! We also congratulate Mr lb Pedersen for his very careful record keeping and for making the decision to place it in the public domain, in the public interest. In the third part of this dossier we have translated a press release relating to the new Danish research project which will examine the effects of GM soy on pigs during the period of weight gain from 30 kg to slaughter at c 110 kg. While we applaud the fact that this research will be conducted, we are concerned that the feeding of the weaned animals from 7 kg (28 days) up to the 30 kg weight will potentially mask GM effects and compromise the results. As the newspaper suggests, it will be in nobody's best interests if these trials are mistrusted or later found to be fraudulent. Dr Brian John from GM-Free Cymru agrees, and says: "We have been involved in GM issues for more than a decade, and we know the score. We can take it as read that there are large sections of the GM industry, and maybe large parts of the farming community, especially in the United States, who will move heaven and earth to prevent anything damaging to the GM cause from seeing the light of day. We suspect that huge pressure has already been put on the Danish journalist and her newspaper by certain interested parties, including farming unions, agrichemical companies and so forth. That is plain stupid of them; their interests are served least of all if real animal welfare and food safety issues are brushed under the carpet." In the fourth part of the dossier we report on an interview with another Danish farmer whose experiences relating to a shift from GM soy animal feed to non-GM soy feed appear to match very closely the experiences of Mr Pedersen. Part 1. The Pilegaarden Findings Animal Health and Welfare When Danish pig farmer lb Borup Pedersen replaced GM soy with non-GM soy in the feeding schedules on his farm, he immediately observed positive changes in the health of the sow herd. He has spoken to a mainstream Danish farming newspaper (Effektivt Landbrug) about this, and the editor has devoted much space to the issue - presumably on the basis that Mr Pedersen is not an organic farmer, but a regular farmer using the same intensive techniques to raise pigs as many other farmers across the country. He farms at Pilegaarden, Hvidsten, near Randers on Jylland. Danish pig production is recognised worldwide for having a the highest level of productivity, with an average of almost 30 weaned pigs per sow per year. Combined with the an antibiotic use of less than 50 mg per kg of produced pork (most other countries except the other Scandinavian states use 2-4 times that amount), that makes them the unofficial "world champions" of swine production. The fact that all use of antibiotics is recorded and strictly controlled by veterinary officers makes statements from the Danish farmer Molecular Ecology(2009)18,569571 NEWS AND VIEWS PERSPECTIVE Unwanted Transgenes Re-Discovered in genetic erosion has been found to date.A similar process could Oaxacan Maize be taking place in Mexico,depending on the dynamics of gene flow via pollen and seeds,yet a more sensitive topic is the ALLISON SNOW unintended spread of transgenes that are banned for cultivation. Department of Evolution,Ecology,and Organismal Biology,Ohio Several types of GM maize are grown in other countries,but State University,Columbus,OH 43210 the key role of landraces in Mexico,which is the centre of origin for this staple crop,has prompted caution for allowing Received 7 November 2008;revision received GM varieties to be cultivated.Some environmental groups,in 26 November 2008;accepted 27 November 2008 particular,view the spread of transgenes as genetic contami- nation,and many people are dismayed by the apparent futility Transgenic [GM (genetically modified)] maize is shipped of efforts to keep transgenes out of landrace gene pools. all over the world, and its ability to germinate, grow, and Monitoring for the presence of transgenes in the Mexican hybridize with local landraces of the crop has generated countryside is a daunting task that requires teams of experts tremendous scientific, social,and political controversy. The from several disciplines(e.g.Serratos-Hernandez et al.2007; great genetic diversity of farmer-produced landraces represents Mercer & Wainwright 2008). Furthermore, the politically a vital resource for subsistence farmers,future crop breeding, sensitive nature of this information has made it difficult for and cultural heritage preservation. Although the Mexican researchers to publish their findings. Here, Pineyro-Nelson government banned GM maize cultivation in 1998, vast et al.(2008)have carried out a masterful job of assembling a quantities of living GM grain are imported from the USA and large data set that will help advance the scientific quality of seeds can easily enter the country by other routes.In 2000, future efforts to monitor the dispersal of GMOs in the environ- Quist&Chapela(2001)discovered transgenes in four ears of ment.Nearly all GM maize varieties have the 35S promoter landrace maize and in seeds from a government-sponsored from the cauliflower mosaic virus,and most have a nopaline Distribution Conasupo Sociedad Anonima (DICONSA) grain synthase (NOS) terminator from Agrobacterium tumefaciens. distribution centre in Oaxaca. Their controversial paper in Thus,these elements are useful markers for other transgenes, Nature set off an explosion of publicity and speculation about and polymerase chain reaction(PCR)-based methods can be used how widely these novel genetic elements had proliferated, to detect a single transgenic seed in a homogenized sample of and what the consequences of ubiquitous gene flow might be. several thousand seeds.However,the exquisite sensitivity of However, a more extensive survey of this region failed to PCR amplification is both a benefit and a liability, as trace detect transgenes in 2003 and 2004(Ortiz-Garcia et a1.2005a), amounts of contamination can yield false-positives.Pineyro- suggesting that transgenic plants were rare or absent in the Nelson et al.used duplicate or triplicate,blind-coded samples sampled fields. Here,Pineyro-Nelson et al. (2008) provide a to address this problem,and positive PCR results from their valuable counterpoint to that survey,resolving apparent con- 2001 samples were confirmed with Southern analyses and tradictions in the literature and raising the bar for subsequent sequencing. studies of immigrating transgenes.They show that transgenes Pineyro-Nelson et al.(2008)found evidence for transgenes were present in Oaxaca in both 2001 and 2004.Their paper in three out of 20 different communities that were sampled in explains how sampling methods, statistical analyses, and 2001 and for 2002(Table 1).When two of the three communities problems with analytical techniques can lead to inconsistent with positive results were sampled more intensively in 2004, estimates of transgene frequencies in maize populations. transgenic plants were detected in 3 of 30 fields in Santiago This is a must-read paper for those who follow genetically Xiacui,and 8 of 30 fields in Santa Maria Jaltianguis.Based on modified organisms(GMO)biosafety research. interviews with farmers about seed exchange,the authors con- Even with non-GM crops,researchers have worried about eluded that transgenes probably persisted in these communities whether the diverse and locally adapted gene pools of landraces after 2001,rather than having been re-introduced.They also could be overwhelmed by abundant gene flow from modern present a simulation model showing that transgenes are likely cultivars.Indeed,another study in this issue of Molecular Ecol- to be highly aggregated geographically when pollen and seed ogy reports a significant amount of introgression from modern mixing are limited,as appears to be the case in these communities. hybrid maize into 'flint' landraces that are preferred for This finding challenges statistical methods for calculating the making polenta in Italy(Bitocchi et aI.2008).Despite several probability of detecting rare events based on assumptions of decades of gene flow into Italian landraces,no evidence for random or uniform geographical distributions(see Cleveland et al.2005;Ortiz-Garcia et al.2005b).Indeed,the patchiness of Correspondence: Allison Snow, Fax: 614-292-2070; E-mail: transgenic plants could explain why none were detected by snow.1 @osu.edu Ortiz-Garcia et al.(2005a),even though they sampled several 0 2009 Blackwell Publishing Ltd Environmental Group Says GM Crops Fail to Tackle Climate Change Date Posted: March 2, 2010 GM Crops Failing to Tackle Climate Change (24 February 2010) -- On the day of the release of annual industry-sponsored figures, a new report from Friends of the Earth International reveals that claims made by the biotech industry that genetically modified (GM) crops can combat climate change are both exaggerated and premature. The report, 'Who Benefits from GM Crops?", examines the evidence for these claims, and exposes that GM crops could actually increase carbon emissions while failing to feed the world. This is because GM crops are responsible for huge increases in the use of pesticides in the US and South America, intensifying fossil fuel use. The cultivation of GM soy to feed factory farmed animals is also contributing to widespread deforestation in South America. The report also exposes that globally GM crops remain confined to less than 3% of agricultural land and more than 99% are grown for animal feed and agrofuels, rather than food. Ongoing concerns about the negative impacts of GM crops means many Governments are still cautious about adopting them. India has placed a moratorium on the planting of its first GM food crop due to widespread concerns on its health, environmental and socio-economic impact. In Europe, the area planted with GM crops has decreased for the fifth year in a row - a reduction of more than 10% since 2008. This reflects continuing public and political concerns on the negative impacts of GM crops. In 2009, the EU's largest member, Germany, became the sixth EU country to ban the planting of GM maize, making the area planted in the EU with GM crops less than 0.05%of total agricultural land. Friends of the Earth Europe GM spokesperson Kirtana Chandrasekaran said, "The number of fields growing GM crops in Europe continues to dwindle while at the same time more and more Europeans are demanding farming that benefits both people and the planet. European Governments would be well advised to steer clear of GM crops in tackling climate change and put their energies into boosting planet and people friendly farming instead." Despite many decades of research there is still not a single commercial GM crop with increased yield, drought-tolerance, salt-tolerance, enhanced nutrition or rt r other beneficial traits long promised by biotech companies. GM crops also hinder the development of real solutions to hunger and climate change by starving them of funding and restricting the access of farmers to seeds and knowledge. Ecological farming and traditional knowledge have been identified as the key to facing future challenges. Friends of the Earth International food coordinator Martin Drago said, "GM crops are being promoted as a solution to climate change, when in reality they are wiping out forests, damaging farmers' livelihoods and increasing harmful emissions. The reality is that GM farming is not a success story. Small farmers across the world are already using planet-friendly methods to feed themselves and cool the planet. These methods must be supported rather than environmentally and socially destructive GM farming." Friends of the Earth Europe campaigns for sustainable and just societies and for the protection of the environment, unites more than 30 national organisations with thousands of local groups and is part of the world's largest grassroots environmental network, Friends of the Earth International. Herbicides found in Human Urine Ithaka Journal 1/2012: 270-272 (2012) www.ithaka-journal.net Editor: Delinat-Institute for Ecology and Climate-farming, CH-1974 Arbaz www.delinat-institut.orq German original here: http://www.ithaka-journal.net/herbizide-im-urin Glyphosate is the main active substance used in most commercial herbicides. It poisons not only plants, but also animals and humans. When testing for glyphosate contamination in an urban population, a German university found significant contamination in all urine samples with levels 5 to 20 times above the legal limit for drinking water. Most herbicides used in commercial agriculture and small gardens as well as for de-weeding railway lines, urban pavements and roadsides contain the active substance glyphosate. The most widely used glyphosate containing herbicides goes under the name "Roundup" by Monsanto. Since the patents on glyphosate have expired, several other agrochemical companies such as Syngenta, Bayer, Nufarm and Dow AgroScience have been producing and selling herbicides containing glyphosate. To date approximately half of the 800,000 tons of glyphosate produced annually worldwide are produced in China. Glyphosate was invented in Switzerland in 1950 and first synthesized by Monsanto in 1970. The compound was found to radically affect the metabolism of plants by preventing them from forming essential amino acids. Glyphosate is a systemic-acting broad[-spectrum] herbicide that kills almost all green plants. Depending on the rate of metabolism, the affected plants die off completely within a few days. Monsanto, Bayer and other companies genetically engineer crops that are resistant to glyphosate. Thus, plantations with genetically modified corn, soy or canola can be sprayed with glyphosate to exterminate weeds between the crop plants. This method, however, results in a glyphosate residue on the crop, which then enters the food chain of animals and humans. , Glyphosate Tolerant Crops Bring Diseases and Death New research reveals disastrous ecological impacts of the world's top herbicide and GM crops made tolerant to it Dr. Mae-Wan Ho and Brett Cherry Dec, 2010 Glyphosate tolerant (GT) crops and glyphosate herbicide (commercial formulation, Roundup) poison nitrogen fixing and other beneficial soil bacteria, increase fungal pathogens, undermine plant immunity to diseases, decrease plant micronutrients available in the soil, and more. Research findings over the past decades paint a damning picture of the cropping system that has taken over 85 percent of the 134 million hectares of global agricultural land now growing genetically modified (GM) crops (see [1] Scientists Reveal Glyphosate Poisons Crops and Soil, SiS 47). The unprecedented rise in GT crops has been accompanied by a sharp increase in the use of the glyphosate herbicides worldwide, especially in the US [2] GM Crops Increase Herbicide Use in the United States, SiS 45). The ecological disaster has been unfolding amid mounting evidence of the herbicide's adverse impacts on human and animal health [3, 4] (Glyphosate Herbicide Could Cause Birth Defects, Ban Glvphosate Herbicides Now, SiS 43), and the breakdown of the Roundup Ready (RR) cropping system as weeds and superweeds become resistant to the herbicide [5, 6] (GM Crops Facing Meltdown in the USA, Glyphosate Resistance in Weeds - The Transgenic Treadmill, SiS 46) [7]. How glyphosate works Glyphosate (N-(phoshonomethyl)glycine) (Figure 1) is a broad-spectrum herbicide initially patented by Monsanto in the 1970s under the trade name Roundup. Figure 1 Glyphosate (N-(phoshonomethyl)glycine) Glyphosate kills plants by binding to and inhibiting the enzyme 5-enolpyruvylshikimate-3- phosphate synthase (EPSPS) of the shikimate pathway for the synthesis of the aromatic amino acids, phenylalanine, tyrosine and tryptophan. These amino acids are essential building blocks for all proteins, and also precursors for growth factors and phytoalexins, compounds involved in the plant's defence against diseases [8, 9]. Animals do not have the shikimate pathway and depend on getting the essential amino acids from their diet. GT plants depend on incorporating an EPSPS from the soil bacterium Agrobacterium tumefaciens (which causes crown gall disease) that is insensitive to glyphosate, and hence not killed by the herbicide. For a long time, glyphosate has been promoted as the safest and most environmentally benign herbicide available. But glyphosate has many other effects that act synergistically on crop health and productivity that extends well beyond the plant into the soil ecosystem and the wider environment. The first hints of these effects came from observations that glyphosate application greatly Sources: Steve Hallett, 765 494-7649, halletts @purdue.edu Bill Johnson, 765-494-4656, wgj @purdue.edu Jessica Schafer, 765-496-6690 , schafer3 @purdue.edu Ag Communications: (765) 494-2722; Keith Robinson, robins89 @purdue.eduAgriculture News Page ABSTRACT Response of Giant Ragweed (Ambrosia trifida), Horseweed (Conyza canadensis), and Common Lambsquarter (Chenopodium album) Biotypes to Glyphosate in the Presence and Absence of Soil Microorganisms Jessica R. Schafer, Steven G. Hallett, and William G. Johnson In previous research conducted on non-weed species, the efficacy of glyphosate was shown to be greater in unsterile soils compared to sterile soils, and that soil microorganisms played an important role in glyphosate efficacy. Conducting greenhouse studies in microbe-free soil may, therefore, produce unreliable data, leading to erroneous conclusions. The objective of this study was to determine the effect of soil microorganisms on the response of glyphosate-resistant and - susceptible biotypes of three problematic weeds of the Midwestern United States: giant ragweed, horseweed and common lambsquarters. A greenhouse dose-response study was conducted on each of the three weed species grown in sterile and unsterile field soil, and the dry weight response of roots and shoots was measured. The three weed species responded differently to glyphosate when grown in the sterile and unsterile soil, that is, in the presence and absence of soil microbes. Soil microbes influenced the response of the susceptible and resistant giant ragweed biotypes and the susceptible common lambsquarters, but not on the tolerant common lambsquarters or either horseweed biotype. The different response of the three species to glyphosate in the presence and absence of soil microbes demonstrates that rhizosphere interactions are fundamental to the mode of action of glyphosate. These findings suggest that the range of tolerance to glyphosate observed in weeds and the evolution of resistance in weed biotypes may also be influenced by rhizosphere interactions. The soil media used in dose-response screenings to identify susceptible and resistant weed biotypes is very important. Unsterile field soil should be incorporated into growth media when conducting dose-response screenings to avoid false positive results. In addition, researchers performing glyphosate dose-response assays should be aware of these findings Impact of Genetically Engineered Crops on Farm Sustainability in the United States THE IMPACT OF GENETICALLY ENGINEERED CROPS ON FARM SUSTAINABILITY IN THE UNITED STATES Committee on the Impact of Biotechnology on Farm-Level Economics and Sustainability Board on Agriculture and Natural Resources Division on Earth and Life Studies NATIONAL RESEARCH COUNCIL OF THE NATIONAL ACADEMIES THE NATIONAL ACADEMIES PRESS Washington, D.C. www.nap.edu Copyright©National Academy of Sciences.All rights reserved. Impact of Genetically Engineered Crops on Farm Sustainability in the United States Contents ABBREVIATIONS AND ACRONYMS xix SUMMARY 1 1 INTRODUCTION 19 Committee Charge and Approach,21 Study Framework,24 Genetically Engineered Traits in Crops,28 Adoption and Distribution of Genetically Engineered Crops,30 Deterrents to Genetically Engineered Trait Development in Other Crops,47 From Adoption to Impact,50 Conclusion,52 References,52 2 ENVIRONMENTAL IMPACTS OF GENETICALLY ENGINEERED CROPS AT THE FARM LEVEL 59 Environmental Impacts of Herbicide-Resistant Crops, 60 Environmental Impacts of Insect-Resistant Crops,83 Gene Flow and Genetically Engineered Crops, 104 Conclusions, 111 References, 112 xiii Copyright©National Academy of Sciences.All rights reserved. Impact of Genetically Engineered Crops on Farm Sustainability in the United States xiv CONTENTS 3 FARM-LEVEL ECONOMIC IMPACTS 135 Economic Impacts on Adopters of Genetically Engineered Crops, 135 Economic Impacts on Other Producers, 164 Socioeconomic Impacts of Gene Flow, 169 Conclusions, 174 References, 175 4 FARM-SYSTEM DYNAMICS AND SOCIAL IMPACTS OF GENETIC ENGINEERING 187 Social Impacts of On-Farm Technology Adoption, 188 Social Networks and Adoption Decisions, 191 Interaction of the Structure of the Seed Industry and Farmer Decisions, 192 Social and Information Networks Between Farmers and Industry, 199 Interaction of Legal and Social Issues Surrounding Genetic Engineering,203 Conclusions,206 References,207 5 KEY FINDINGS,REMAINING CHALLENGES,AND FUTURE OPPORTUNITIES 213 Key Findings,214 Remaining Challenges Facing Genetically Engineered Crops,216 Future Applications of Genetically Engineered Crops,219 Research Priorities Related to Genetically Engineered Crops,227 Advancing Potential Benefits of Genetically Engineered Crops by Strengthening Cooperation Between Public and Private Research and Development,229 References,232 APPENDIXES A Herbicide Selection 237 B Tillage Systems 245 C Biographical Sketches of Committee Members 247 Copyright©National Academy of Sciences.All rights reserved. Illegal gene flow from transgenic creeping bentgrass: the saga continues by Dr Allison Snow Molecular Ecology, Volume 21 , Issue 19, pages 4663-4664, October 2012 DOI: 10.1111/j.1365-294X.2012.05695.x ABSTRACT: Ecologists have paid close attention to environmental effects that fitness-enhancing transgenes might have following crop-to-wild gene flow (e.g. Snow et al. 2003). For some crops, gene flow also can lead to legal problems, especially when government agencies have not approved transgenic events for unrestricted environmental release. Creeping bentgrass (Agrostis stolonifera), a common turfgrass used in golf courses, is the focus of both areas of concern. In 2002, prior to expected deregulation (still pending), The Scotts Company planted creeping bentgrass with transgenic resistance to the herbicide glyphosate, also known as RoundUp®, on 162 ha in a designated control area in central Oregon (Fig. 1). Despite efforts to restrict gene flow, wind-dispersed pollen carried transgenes to florets of local A. stolonifera and A. gigantea as far as 14 km away, and to sentinel plants placed as far as 21 km away (Watrud et al. 2004). Then, in August 2003, a strong wind event moved transgenic seeds from windrows of cut bentgrass into nearby areas. The company's efforts to kill all transgenic survivors in the area failed: feral glyphosate-resistant populations of A. stolonifera were found by Reichman et al. (2006), and 62% of 585 bentgrass plants had the telltale CP4 EPSPS transgene in 2006 (Zapiola et al. 2008; Fig. 2). Now, in this issue, the story gets even more interesting as Zapiola & Mallory-Smith (2012) describe a transgenic, intergeneric hybrid produced on a feral, transgenic creeping bentgrass plant that received pollen from Polypogon monspeliensis (rabbitfoot grass). Their finding raises a host of new questions about the prevalence and fitness of intergeneric hybrids, as well as how to evaluate the full extent of gene flow from transgenic crops. y... • GE Crop Risk Assessment Challenges: An Overview BY DR.CHARLES BENBROOK I MAY 6, 2013 There have been dramatic changes in the transgenic composition of GE corn and soybeans over the last five years, coupled with a substantial increase in reliance on pesticides and Bt toxins. Compared to the first five years of commercial use (1996-2000), today's GE corn and soybeans in the U.S. require: About twice as much herbicide per acre, with glyphosate/Roundup accounting for essentially all the growth; In corn, two to six Bt toxins to deal with European corn borer and the corn rootworm complex; Delayed release, systemic seed treatments including at least two insecticides and two fungicides, one of which is a nicotinyl implicated in honey bee Colony Collapse Disorder; A return to corn soil insecticide use as a component of Bt-gene, resistance-management programs (eroding a portion of the reduction in insecticide use brought about by Bt corn); Significant and historically unprecedented increases in fungicide use on corn (ii percent of crop acres were treated in latest USDA pesticide use survey [2010], no more than 1 percent was treated previously); and Approval and commercial planting in the U.S. of the first GE crop that will be consumed in significant quantities by humans in a largely unprocessed form — Bt and RR sweetcorn. The move to stacked varieties expressing multiple traits, coupled with the above changes in the intensity of chemical use required to bring GE crops to harvest, raises new questions about new routes of exposure and about cumulative levels of exposure to GE proteins, potential allergens and pesticides, especially via drinking water, certain foods made from corn or soybeans, and, for infants, breast milk, cow's milk, soymilk, and formula. It also raises new testing challenges arising from the likely presence of multiple transgenes, DNA fragments, promoters, regulatory sequences and chemicals from pesticides (active ingredients, metabolites, surfactants, adjuvants, etc). s These changes pose serious risk assessment challenges that are, for the most part,being ignored by the industry and regulatory authorities. New information is essential to convince regulators that they must invest substantially more public resources in the independent testing of GE crop safety. The two core goals for a new testing initiative should be to (a) resolve lingering uncertainty over the safety of the GE traits currently on the market, and (b) develop advanced testing methods and protocols for application in the testing of future GE food traits. At a minimum, the following steps should be taken to explore key questions about today's GE crops: 1. Quantification of the levels of pesticides and their metabolites/breakdown products associated with GE crops in key foods and human fluids (blood and urine), encompassing initially glyphosate, AMPA, nicotinyl seed treatments and Bt proteins. Top priority foods to test include whole wheat grain and flour, whole wheat bread, and soy-derived ingredients and foods. Essential liquids to test include cow's milk, breast milk, soymilk and infant formula. 2. Development of methods to accurately quantify GE protein exposure levels in tissues and organs of concern in evaluating human health risks. The research would also be designed to track the breakdown products formed as GE proteins move through the digestive system. Methods would be developed to identify the form of novel proteins or other phytochemicals from GE foods, as well as the quantities passing from the GI tract into the bloodstream, from the blood to the kidneys and liver, and in the case of pregnant women, across the blood-brain barrier. 3. Methods must be developed to assess the impacts on fetal development following pre-natal exposure to GE proteins via maternal blood flows. Focus should be on epigenetic patterns of gene expression and the frequency of auto-immune diseases including food allergies, asthma and behavioral problems. 4. In light of novel combinations of exposures, including combinations of glyphosate, AMPA, a nicotinyl insecticide and Bt proteins, short-term cell assays should be used to test for toxic potential of each of these singly and in various combinations. The results can be used to target subsequent, long-term testing. • 5. An appropriate, government agency or international organization should fund long-term toxicology and cancer feeding studies in at least two species of laboratory animals on a cross-section of the major traits now in GE varieties. Trials should cover the most widely planted Bt endotoxins alone, the EPSPS gene conferring resistance to glyphosate alone and these two traits in combination. At the conclusion of a first round of testing, the research team should issue recommendations for the design and conduct of future cancer feeding trials applied to GE food traits and render judgments regarding the need for additional testing. Do escaped transgenes persist in nature? The case of an herbicide resistance transgene in a weedy Brassica rapa population To cite this article: S. I. WARWICK, A. LEGERE, M.-J. SIMARD, T. JAMES Molecular Ecology 2007 Oct 29 2007; (advanced online pre-publication) [authors details at end] http://wwvv.blackwell-synercw.com/doi/abs/10.1111/.1365- 294X.2007.03567.x Keywords: Brassica napus, Brassica rapa, gene flow, genetically modified crops, hybridization, transgene escape Abstract The existence of transgenic hybrids resulting from transgene escape from genetically modified (GM) crops to wild or weedy relatives is well documented but the fate of the transgene over time in recipient wild species populations is still relatively unknown. This is the first report of the persistence and apparent introgression, i.e. stable incorporation of genes from one differentiated gene pool into another, of an herbicide resistance transgene from Brassica napus into the gene pool of its weedy relative, Brassica rapa, monitored under natural commercial field conditions. Hybridization between glyphosate-resistant [herbicide resistance (HR)] B. napus and B. rapa was first observed at two Quebec sites, Ste Agathe and St Henri, in 2001. B. rapa populations at these two locations were monitored in 2002, 2003 and 2005 for the presence of hybrids and transgene persistence. Hybrid numbers decreased over the 3-year period, from 85 out of -200 plants surveyed in 2002 to only five out of 200 plants in 2005 (St Henri site). Most hybrids had the HR trait, reduced male fertility, intermediate genome structure, and presence of both species-specific amplified fragment length polymorphism markers. Both Fl and backcross hybrid generations were detected. One introgressed individual, i.e. with the HR trait and diploid ploidy level of B. rapa, was observed in 2005. The latter had reduced pollen viability but produced -480 seeds. Forty-eight of the 50 progeny grown from this plant were diploid with high pollen viability and 22 had the transgene (1:1 segregation). These observations confirm the persistence of the HR trait over time. Persistence occurred over a 6-year period, in the absence of herbicide selection pressure (with the exception of possible exposure to glyphosate in 2002), and in spite of the fitness cost associated with hybridization. w Agriculture and Agri-Food Canada (AAFC), Eastern Cereal and Oilseeds Research Center, Central Experimental Farm, Ottawa, ON, Canada K1A 006. S. I. WARWICK* *Agriculture and Agri-Food Canada (AAFC), Eastern Cereal and Oilseeds Research Center, Central Experimental Farm, Ottawa, ON, Canada K1A 006, , A. LEGEREt tAAFC-Saskatoon, 107 Science Place, Saskatoon, SK, Canada S7N 0X2, , M.-J. SIMARD$ $AAFC-CRDCGC, 2560 Boul. Hochelaga, Quebec, QC, Canada G 1 V 2J3 and T. JAMES* *Agriculture and Agri-Food Canada (AAFC), Eastern Cereal and Oilseeds Research Center, Central Experimental Farm, Ottawa, ON, Canada K1A 006, *Agriculture and Agri-Food Canada (AAFC), Eastern Cereal and Oilseeds Research Center, Central Experimental Farm, Ottawa, ON, Canada K1A OC6, tAAFC-Saskatoon, 107 Science Place, Saskatoon, SK, Canada S7N 0X2, tAAFC-CRDCGC, 2560 Boul. Hochelaga, Quebec, QC, Canada G1V 2J3 Correspondence: Dr Suzanne Warwick. Fax: (+01) 613 759-1701; E-mail: warwicks(c�agr.gc.ca