Please see the EU relevant links below.

Event specific real-time quantitative PCR based method for genetically modified maize SYN-BTØ11-1xMON-ØØØ21-9. - Validated by the Community reference laboratory established under Regulation (EC) No 1829/2003. Please see the EU relevant links below. -- Reference Material: ERM®-BF412 (for SYN-BTØ11-1) accessible via the Joint Research Centre (JRC) of the European Commission, the Institute of Reference Materials and Measurements (IRMM) at http://www.irmm.jrc.be/html/reference_materials_catalogue/index.htm and AOCS 0407 (for MON-ØØØ21-9 ) accessible via the American Oil Chemists Society at http://www.aocs.org/tech/crm

Please see the EU relevant links below.

Method for detection: Event specific real-time quantitative PCR based methods for SYN-BTØ11-1, SYN-IR162-4, SYN-IR6Ø4-5 and MON-ØØØ21-9 maizes; the detection methods are validated on the single-trait events and verified on genomic DNA extracted from seeds of SYN-BTØ11-1 × SYN-IR162-4 × SYN-IR6Ø4-5 × MON-ØØØ21-9 maize. Reference material: ERM®-BF412 (for SYN-BTØ11-1) and ERM®-BF423 (for SYN-IR6Ø4-5) accessible via the Joint Research Centre (JRC) of the European Commission, and AOCS 1208-A and AOCS 0407-A (for SYN-IR162-4), AOCS 0407-A and AOCS 0407-B (for MON-ØØØ21-9) accessible via the American Oil Chemists Society.

The stacked event Bt11 x GA21 maize confers tolerance to lepidopteran insects, especially to Diatraea saccharalis, and tolerance to herbicides which active principle is glyphosate and glufosinate ammonium.
Both parental events, Bt11 and GA21, were stacked by conventional crossing (sexual). The stacked event has the mepsps gene from the GA21 event and the genes cry1Ab and pat from the Bt11 event. The transgenes are inherited in independent form, since it presents mendelian segregation. Moreover, the applicant proved the gene stability and the effective levels of the expressed proteins.
The protein Cry1Ab confers resistance to certain lepideroptean insects, and proteins PAT and mEPSPS allows tolerate herbicides ammonium glufosinate and glyphosate respectively.
After comparison of 65 analytes measured in grain and forage, in the compositional analysis study, it is concluded that maize Bt11 x GA21 is equivalent to commercial maize and the parental lines.
The allergenicity and toxicity assessment of the new expression proteins were carried out previously in both single events.
Taking into account the assessment of genetic stability, molecular characterization, products and levels of expression, compositional analyses and morphoagronomic studies, no metabolic interaction is expected that might impact on the food safety when single events are stacked in a conventional way.
The Bt11xGA21 event is substantial and nutritionally equivalent to its non transgenic counterpart.

Please see decision document weblinks

commercial release of genetically modified insect resistant and herbicide tolerant corn Bt11 x GA21

Bt11 x GA21 corn was generated by classic genetic development, through sexual crossing between genetically modified lineages containing either event BT11 or GA2, previously approved by the National Biosafety Technical Commission – CTNBio to be commercially released. Bt11 x GA21 corn was already approved in the United States, Canada, Japan, Mexico, Korea and the Philippines for human and animal consumption. In the process of developing Bt11 x GA21 corn there were no genetic modifications besides the introgression of both genes in corn lineages. Compared to conventional corn, Bt11 x GA21 corn has no higher ability to survive as a pest. The presence of genes granting resistance to Lepidoptera insects and tolerance to the glyphosate insecticide give Bt11 x GA21 corn selective advantage when exposed to the herbicide and in the presence of target insects. However, these features are not sufficient to turn the product into a pest in corn cultures. The use of corn containing the stacked event, Bt11 x GA21, represents a future trend meeting corn producers’ demand – by combining two important agronomic features into a same hybrid plant. Event Bt11 contains gene cry1A(b) – coming from Bacillus thuringiensis, granting resistance to certain insects, and gene pat, derived from Streptomyces viridochromogenes, a soil bacterium used as a selection marker during the transformation process. Gene cry1A(b) is responsible for producing protein Btk that is proteolytically cleavated in the alkaline intestine of Lepidoptera insects in an active insecticide form. This insecticide protein, when active, interacts with a receptor molecule that is present solely in the epithelial cells of the middle intestine of susceptible insects, generating pores in cell membranes. When the pores are formed, the cell osmotic balance becomes disrupted, cells swell and undergo lysis. The larvae of susceptible insects, when submitted to the Cry protein stop feeding and later die. Several bonding sites featuring high affinity for Bt proteins were already identified in the middle intestine of susceptible insects. Studies showed that the insecticide protein codified by gene cry1A(b) is highly specific for Lepidoptera insects. Event GA21 contains the mutant gene mepsps that is responsible for the expression of protein 5 enolpyruvyl-shikimate-3-phosphate-syntase (mEPSPS). The EPSPS protein is a key enzyme in processing the shikimic acid, which is involved in the biosynthesis of aromatic amino acids and is commonly found in plants, fungi and bacteria. The enzyme does not exist in animals. The EPSPS enzyme is highly sensitive to herbicide products containing the glyphosate active principle. Results from hybridation data show that Bt11 x GA21 corn maintains its hybridation standard in the same way as the respective parental events for each corresponding transgene. The set of evidence obtained based on results of comparative molecular analysis, analysis of genetic heritage standards and comparative analysis of level of expression for proteins Cry1A(b) and mEPSPS in Bt11 x GA21 corn suggest that the levels of exposure to non-target organisms and in human and animal feeding are the same as to either Bt11 or GA21 measured individually. The presence of genes pat and mepsps make the Bt11 x GA21 corn resistant to a pair of chemical products with herbicide properties available in the market: glyphosate and gluphosinate ammonium. Should corn have potential to turn into a pest plant, its control where such products were used could increase its invasive ability. However, the high degree of domestication of corn made the species highly dependent on man. This may be ratified by the absence of pest or even feral populations in agricultural and natural environments, even after millennia of cultivation in Brazil. Therefore, the risk that corn changes into a pest plant, if such risk exists, is negligible. According to Annex I to Ruling Resolution nº 5, of March 12, 2009, the applicant shall have a term of thirty (30) days from the publication of this Technical Opinion to adjust its proposal to the post-commercial release monitoring plan. Under Article 14 of Law no. 11,105/2005, CTNBio found that the request complies with the applicable rules and legislation securing the biosafety of environment, agriculture, human and animal health and reached a conclusion that the stacked corn Bt11 x GA21 is substantially equivalent to conventional corn and its consumption is safe for human and animal health. Regarding the environment, CTNBio’s conclusion is that cultivation of Bt11 x GA21 corn is not a potential cause of significant environmental degradation, keeping with the biota a relation identical to that of conventional corn. TECHNICAL OPINION I. GMO Identification GMO name: Bt11 x GA21 corn. Applicant: Syngenta Seeds Ltda. Species: Zea Mays L. Inserted characteristics: Tolerance to glyphosate herbicide and insect resistance Method of insertion: Bt11 x GA21 corn, ranked as Risk Class I, was developed by classical genetic improvement, through sexual crossing between genetically modified lineages containing event Bt11 and event GA21. Prospective use: Free registration, use, essays, tests, sowing, transport, storage, marketing, consumption, import, release and discarding. II. General Information Corn, Zea Mays L. is a species of the family Gramineae, tribe Maydae, sub-family Panicoidae that is separated within the sub-genus Zea and has chromosome number 2n = 20,21,22,24(1). The wild species closest to corn is the teosinte found in Mexico and some regions of Central America, where it is able to cross with cultivated corn in the production fields. Corn has a history of over eight thousand years in the Americas, and is cultivated since the pre-Columbian era. Among higher plants, corn is the best scientifically characterized and is currently the cultivated species that reached the highest degree of domestication and is unable to survive in nature but when cultivated by man(2). There are currently over 300 identified races of corn and, within each such race, thousands of cultivars. One of the most important sources of food in the world, corn is an input in the production of a wide range of foodstuff, rations and industrial products. Brazil is one of the largest producers of corn over the world, and corn is cultivated nearly all over the national territory(3). Occurrence of insects in Earth is larger in the tropics than in temperate regions, where the damages caused by such animals are more noticeable. Among the most damaging corn pests an important place is taken by the fall armyworm, Spodoptera frugiperda. Cruz et al. (4) estimated that the losses in Brazil caused by infestations of S. frugiperda reach 400 million Dollars each year. Other species of the order Lepidoptera are also important pests in the culture of corn, such as the corn earworm (Helicoverpa zea) and stalk borer (Ditraea sacharalis). The main measure to control insects in the corn cultivation has been the use of insecticides. In some areas of the Brazilian Center-West, for instance, tenths of insecticide sprays are needed in a single culture cycle. Another measure to control pests would be the use of resistant cultivars. Bt11 x GA21 corn was developed through classic genetic improvement, by sexual crossing between genetically modified lineages containing separately event Bt11 and event GA21(5,6). Bt11 x GA21 corn was already approved in the United States, Canada, Japan, Korea and the Philippines for animal and human consumption. In the course of developing Bt11 x GA21 corn, there were no other genetic changes in addition to the introgression of both events contained in corn lineages Bt11 and GA21 separately(7). Compared with conventional corn, Bt11 x GA21 corn fails to display greater ability to survive as a pest. The presence of gens granting resistance to Lepidoptera insects and tolerance to glyphosate herbicide give a selective advantage to Bt11 x GA21 corn when it is exposed to the herbicide and submitted to the presence of target-insects. However, such characteristics are not sufficient to make this corn a pest in corn cultivars(8). The use of corn featuring the stacked event, Bt11 x GA21, represents a future trend – that meets the demand of farmers – by combining two agronomically important features in a single hybrid. Corns with events combined by classic genetic improvement are approved in Japan, Korea, Philippines and are under analysis in a number of other countries(7). III. Description of GMO and Proteins Expressed The corn containing event Bt11 was obtained by direct transfer of DNA in protoplasts of corn lineage H8540 by inserting plasmid pZO1502 containing genes cry1Ab and pat. The expression product of gene cry1Ab is protein Cry1Ab that has insecticide activity on target-pests, protecting the plants from damages caused by such pests. Gene cry1Ab was isolated from bacterium B. thuringiensis subspecies kurstaki strain HD-1(3,8,9). Gene pat is derived from Streptomyces viridochromogenes strain Tu494 and is responsible for codifying enzyme phosphinothricin N-acetyltransferase (PAT). This enzyme is able to chemically inactivate herbicides derived from phosphinotricin, such as glyphosinate ammonium, making resistant cells and plants containing the enzyme. The PAT enzyme activity has its activity described and well known(9). Event GA2 contains gene mepsps that expresses enzyme 5-enolpyruvyl-shikimate-3-phosphate-synthase (mEPSPS). Protein mEPSPS differs from feral EPSPS in two amino acids and is a key enzyme in the shikimic acid process, involved in the biosynthesis of aromatic amino acids. mEPSPS is highly sensitive to herbicide products containing glyphosate. Corn plants transformed with mEPSPS gene, such as those derived from event GA2, synthesize protein mEPSPS that grants tolerance to herbicide products containing glyphosate(10,11). Proteins mEPSPS and Cry1Ab mode of action and biologic activities expressed in Bt11 x GA21 corn are different and do not posses known interaction mechanisms that are able to cause adverse effects to human and animal health, and to the environment. mEPSPS and Cry1Ab proteins present in Bt11 x GA21 corn are accumulated in different cell compartments and display distinct and non-interactive metabolic functions. Therefore, protein mEPSPS is directed to the chloroplast while protein Cry1Ab is accumulated in the cytoplasm(5,6). Expression level of proteins mEPSPS and Cry1Ab is low in individual events (GA21 corn and Br11 corn) and therefore the likelihood that such proteins would interact between them is held improbable, a fact that is macroscopically confirmed through the analysis of agronomic and phenotypical characteristics related to efficacy and selectivity of Bt11 x GA21 corn in the fields(5,6). IV. Aspects Related to Human and Animal Health Safety aspects of proteins Cry1Ab and EPSPS were thoroughly assessed by CTNBio and protein Cry1Ab mode of action is well clarified by scientific literature(5,6). In vitro tests were used to assay increased digestibility of foodstuffs containing pre-heated proteins Cry1Ab and mEPSPS. The study showed that pre-heating increases the protein digestibility in simulated gastric and intestine fluids, suggesting that the likelihood of an eventual allergenic potential of protein Cry1Ab is extremely low, for the easy of its digestion(5,6). Further, in vivo and in vitro studies confirmed that proteins Cry1Ab expressed in B. thuringiensis and Bt11 x GA21 corn are highly selective and do not act on mammals(13,14,15,16,17,18,19). Protein mEPSPS is an enzyme that is present in all plants and in a large number of microorganisms(17), while protein Cry1Ab does not display enzymatic activity in plants and therefore fails to affect plant metabolism. The likelihood that biochemical interaction takes place between proteins mEPSPS and Cry1Ab in the complex matrix of a plant is limited, since such proteins accumulate in different places of the cells and in a low level of expression. With this, a potential exposure to such proteins is extremely low in human and animal feeding. Considering that proteins mEPSPS and Cry1Ab fail to produce toxicity in the maximum doses tested, it is highly unlikely that an interaction able to cause additive or synergic effects occurs between such proteins in the normal doses found in foodstuffs. The literature in the area of toxicology of chemical mixtures provides information showing that such interactions are inexistent when the substances are administered in doses substantially below the levels of unobserved adverse effect(20,2,22,23). Due to the rigorous specificity for substrates, enzymes EPSPS link just S3P, PEP and glyphosate. The only known metabolic product is the 5-enolpyruvyl shikimic-3-phosphate, which corresponds to the penultimate product of the shikimic acid pathway. Shikimic acid is a precursor for biosynthesis of amino acids (phenylalanine, tyrosine and triptophane) and a number of secondary metabolits, such as tetrahydrofolate, ubiquinone and K vitamin(24). Though the shikimic acid (or shikimate) pathway and proteins EPSPS do not occur in mammals, fish, birds and insects, they are important to plants. It is reckoned that aromatic molecules, all of them derived from shikimic acid, represent no less than 35% of a plant’s dry weight(25,26). In vitro assays performed with simulated digestive fluids are widely used tools as a model for animal digestion. This simulated system was used to probe into digestibility of plant proteins(27,28), animal proteins(29), and food additives(30), as well as to assay the protein quality(31) and the allergenicity potential through absorption of the proteins by the digestive system(32). The knowledge on the mode of action, specificity and safe use history of protein EPSPS, potential toxic and allergenic effects of such proteins to humans and other mammals were assayed through in vitro digestion tests. The studies used simulated gastric fluids (pH 1.2) and intestinal fluids (pH 7.5). The degradation rate of protein mEPSPS (mature protein with no transit peptide) was assessed through Western blot analyses. The study showed that protein mEPSPS and peptides degraded in less than 15 seconds after exposed to the gastric fluid. In the simulated intestinal fluid, degradation of protein mEPSPS occurred in a period shorter than 10 minutes(33). Finally, enzyme EPSPS expressed in corn containing event GA21 has no typical characteristics of known allergens, since the behavior of allergenic proteins in the digestive tract is well described(34,35,36). There are no homology regions when the introduced sequence is compared with known allergen sequences. Besides, several alimentary allergens are known to be stable under heat. V. Environmental Aspects Corn is a monoic, allogamic and annual plant, pollinated mostly by the wind. Distances that may be covered by the pollen depend on wind patterns, humidity and temperature. Corn pollen is freely dispersed close to the cultivated area, and may reach styli-stigmas of the same or different genotypes and, under adequate conditions, starts its germination, generating the pollinic tube and promoting the ovule fecundation within an average term of 24 hours. Studies on pollen dispersion have been conducted and some of them show that pollen may travel long distances, though the majority is deposited close to the cultivated area with a very low translocation rate. Over 95% of the pollen may reach distances within 60 m of the pollen source(37). Luna et al. (38) investigated pollen isolation distance and control, where it was shown that crossed pollination occurs within 200 m. However, no crossed pollination, under conditions of non-detasseling, was noticed in distances higher than 300 m from the pollen source. The results indicate that pollen viability is maintained for two hours and that crossed pollination was not observed in distances of 300 m from the pollen source. When compared with concentrations at one meter from the source culture under low-to-moderate winds it was estimated that about 2% of the pollen reaches 60 meters, 1.1% reaches 200 meters, and 0.75% to 0.5% reaches a distance of 500 meters. At a distance of ten meters from the field, the number of pollen grains per unit of area is tenfold lower than the number observed at one meter from the border. Therefore, if established separation distances developed for the production of corn seeds are observed, it may be expected that pollen transfer to surrounding varieties is minimized and that the presence of glyphosate-tolerant genetic materials is unlikely. There are no kindred species of corn naturally distributed within Brazil. However, though the gene flow to local varieties of open pollination is possible, it poses the same risk than the one caused by commercial genotypes available in the marketplace. In the specific case of crossing between GA21 corn and creole varieties, selected pressure from the management by small farmers is not expected; the transgene will not be incorporated to the genome of creole varieties because in practice a small farmer does not use herbicides. From the agronomic viewpoint, coexistence between cultivars of conventional corn (improved or creole) and transgenic corn is possible(39,40). Old communities and modern farmers have learned how to live on without problems with different corn cultivars, while keeping their genetic identities along time. The likelihood of a transgenic plant becoming an invading species, as well as the likelihood that the crossing of GA21 corn with other corn plants generating an invading plant is negligible, in view of the biologic characteristics of the species and the fact that corn cannot survive without human intervention, a result of the selection made during the plant evolution. Corn is held as the species reaching the highest degree of domestication among cultivated plants, and has lost its ability to survive in nature, such as elimination of thrashing. Therefore, corn is a plant that is unable to survive under natural conditions without technical assistance. Therefore, one expects GA21 corn to exhibit an environmental behavior similar to ordinary corn, being hence negligible the possibility of changing into an invading or pest plant. Introduction of gene elements did not change the plant reproductive features, and the odds of crossed fecundation between hybrids and conventional corn lineages will persist between event Bt11 x GA21 and other corn plants. Gene flow in corn may take place through transfer of pollen and dispersion of seeds – which is easily controlled – since corn domestication eliminated the ancient mechanisms of seed dispersion and the movement of pollen is the only effective form for genes to escape from corn plants. The likelihood that a gene mepsps of a transgenic plant passes over to other organisms, such as, for instance, soil microorganisms, is practically zero(41,42). Naturally, gene epsps is common in plants, fungi and microorganisms, occurring abundantly in nature and does not results in significant risk to the soil microbiota. Besides, there is no evidence that plant genes have in some way been transferred to bacteria under natural conditions. Agronomic parameters and efficacy of controlling pest Lepidoptera in Bt11 hybrid corn were compared with isogenic lineages in assays conducted in five locations: City of Uberlândia, State of Minas Gerais; City of Ituiutaba, State of Minas Gerais; City of Iraí de Minas, State of Minas Gerais; City of Campo Mourão, State of Paraná; and City of Pinhalzinho, State of Santa Catarina. Plant height, ear insertion height, date of male and female flowering, percentage of erect plants, type and color of seed-corn, humidity content, yield and ear damage were the parameters observed in agronomic assays. In order to assay the efficacy of event Bt11 in controlling pest Lepidoptera, damages of fall armyworm (S. frugiperda); corn borer (D. saccharalis); and corn earworm (H. zea) (5,6) were examined. Hybrids containing event Bt11 were efficient to control the assayed pest Lepidoptera and exceeded the agronomic parameters related to corn-seed and grain damage. According to available information, the favorable performance difference was mainly related to the efficient protection against attacks from pests studied. As for other agronomic parameters assayed, Bt11 hybrids exhibited a performance statistically similar to the respective isogenic non-genetically modified hybrids. The results confirm the equivalence of agronomic performance between Bt11 hybrids and isogenous corn non-genetically modified under Brazilian cultivation conditions(5,6). Frequent use of chemical insecticides contributes towards environment degradation, environmental pollution and disruption of the whole ecosystem in the corn culture and even of other cultures in rotation. With the adoption of insect resistant genetically modified plants, reduction of insecticides was considerable in countries where the technology has been used for over ten years. In 2001 alone, United States farmers, for instance, obtained a reduction of over 8,000 tons of active insecticide ingredient in (43,44,45). In China, the use of insecticides were reduced by 67% on average, and the reduction in terms of active insecticide ingredient reached 80%(46). In South Africa, the reduction was about 66%(42). These examples suggest that the use of Bt technology in Brazil may contribute for a diminished use of insecticides and, therefore, reduction of the impacts resulting from the use of such pesticides to the environment and human and animal health. Besides, the use of Bt technologies may have positive effects in preserving populations of non-target organisms and beneficial insects, making an integrated management of crop pests easier(47,48,49). Besides, the use of technologies that reduce spraying chemicals on crops may bring secondary benefits with the reduced use of raw-material in the production of pesticides, reduction of fuels used to produce, distribute and apply pesticides and elimination of the need to use and discard pesticide packaging(50). Compared with conventional corn, Bt11 x GA21 corn has no higher ability to survive as a pest. The presence of genes for Lepidoptera insect resistance and glyphosate herbicide tolerance imparts a selective advantage to Bt11 x GA21 corn when exposed to the herbicide and in the presence of target insects. However, such features are not enough for Bt11 x GA21 corn change into a pest to corn cultures(8). Comparison of amino acid sequences of the new proteins expressed by Bt11 x GA21 corn against sequences of toxic or allergenic proteins show that there is no homology likely to indicate similarities. Therefore, toxic or allergenic effects are not expected as a result of contact or consumption of foodstuffs containing event Bt11 x GA21(8). VI. Restrictions to the Use of the GMO and its Derivatives As established by Article 11 of Law nº 11,460, of March 21, 2007 “research and cultivation of genetically modified organisms may not be conducted in indigenous lands and areas of conservation units.” The studies submitted by applicant showed that there was no significant difference between corn hybrids derived from unmodified lineages and Bt11 x GA21 corn regarding agronomic characteristics; reproduction and dissemination modes; and survival ability. All evidences submitted with the application documents and bibliographic references confirm the risk level of the transgenic variety as being equivalent to the risk level of non transgenic varieties regarding soil microbiota, other plants and human and animal health. Therefore, cultivation and consumption of Bt11 x GA21 corn are not potential causes of significant degradation of the environment, nor of risks to human and animal health. For these reasons, there are no restrictions to the use of such corn and its derivatives, except in locations mentioned by Law nº 11,460, of March 21, 2007. Vertical gene flow to local varieties (the so-called creole corns) of open pollination is possible and poses the same risk as the one caused by commercial genotypes available in the market (80% of conventional corn planted in Brazil comes from commercial seeds that underwent a genetic improvement process). Coexistence of conventional corn (either improved or creole) cultivars and transgenic corn cultivars is possible from the agronomic viewpoint(39,40) and shall comply with the provisions of CTNBio Ruling Resolution nº 4. After being used for ten years in other countries, no problems were detected to human and animal health and the environment that may be ascribed to transgenic corns. It shall be emphasized that the lack of negative effects resulting from farming transgenic corn plants is far from a guarantee that such problems may not occur in the future. Zero risk and absolute safety do not exist in the biologic world, although there is a significant amount of reliable scientific information and a safe history of ten years underlying the fact that Bt11 x GA21 corn is as safe as the traditional corn versions. Therefore, applicant shall conduct the post-commercial release monitoring according to CTNBio Ruling Resolution nº 3. VII. Consideration on the Particulars of Different Regions of the Country (Information to supervisory agencies) As established by Article 11 of Law nº 11,460, of March 21, 2007 “research and cultivation of genetically modified organisms may not be conducted in indigenous lands and areas of conservation units.” VIII. Conclusion Whereas the corn (Zea mays) variety Bt11 x GA21 belongs to a well characterized species with a solid history of safety for human consumption and that genes cry1A(b), pat, mepsps introduced in this variety codify proteins that are ubiquitous in nature and are present in plants, fungi and microorganisms that are part of human and animal alimentary diet; Whereas insertion of this genotype took place through classic genetic improvement and resulted in insertion of a stable and functional copy of cry1A(b), pat, mepsps genes that granted to the plants tolerance to glyphosate herbicide and resistance to insects; Whereas data on centesimal composition failed to show significant differences between genetically modified and conventional varieties, suggesting a nutritional equivalence between them; Whereas CTNBio conducted a separate assay on the events and issued an opinion favorable to commercial release of the separate events; Whereas: 1. Bt11 x GA21 corn is a genetically modified product, displaying resistance to a number of Lepidoptera pests and tolerance to glyphosate herbicide, developed through classic improvement by sexual crossing between lineages containing event Bt11 and event GA21, previously approved for commercial release; 2. Comparative molecular analysis of Bt11 x GA21 corn evidenced that integrity of inserts was maintained during the classic improvement with the purpose of combining both events; 3. Segregation analysis and genetic heritance standards of Bt11 x GA21 corn showed that genes of events Bt11 and GA21 are independent and segregate on a stable manner along successive generations; 4. Agronomic and efficacy assays of Bt11 x GA21 corn indicate that combination of such events by classic genetic improvement methods (sexual crossings) did not lead to expression of any other characteristics, except those already expected, that is to say, resistance to certain insects and tolerance to glyphosate herbicide; 5. Expressions of proteins Cry1Ab and mEPSPS in Bt11 x GA21 corn are not significantly different from their expression in corns containing the separate events; Therefore, considering internationally accepted criteria in the process of analyzing risks in genetically modified raw-material it is possible to conclude that Bt11 x GA21 corn is as safe as its conventional equivalents. In the context of the competences granted to it under Article 14 of Law nº 11,105/05, CTNBio considered that the request complied with the rules and legislation in effect that intend to guaranty environmental and agricultural biosafety and human and animal health, reaching a conclusion that Bt11 x GA21 corn is substantially equivalent to conventional corn, being its consumption safe for human and animal health. Regarding the environment, CTNBio’s conclusion was that the Bt11 x GA21 corn is not a potential cause of significant degradation to the environment, keeping with the biota a relation identical to that of conventional corn. CTNBio advocates that this activity is not a potential cause of significant degradation to the environment or of harm to human and animal health. Restrictions to the use of the GMO analyzed and its derivatives are conditioned to the provisions of Law nº 11,460, of March 21, 2007, and to CTNBio Ruling Resolution nº 03 and Ruling Resolution nº 04. CTNBio assay took into consideration opinions issued by the Commission members; ad hoc consultants; documents delivered to CTNBio Executive Secretary by applicant; results of planned releases to the environment; and discussions, lectures and papers related to the public hearing held on 03.20.2007. Besides, independent studies and scientific literature of applicant, conducted by third parties were also considered. According to Annex I to Ruling Resolution nº 5, of March 12, 2009, the applicant shall have a term of thirty (30) days from publication of this Technical Opinion to adjust its proposal to the post-commercial release monitoring plan. IX. Bibliography 1. Food and Agriculture Organization of the United Nations / World Health Organization. FAO/WHO – 2000a. Grassland Index. Zea mays L. (Disponível em: http://www.fao.org/WAICENT/faoinfo/agricult/agp/agpc/doc/gbase/data/pf000342.htm). 2. BAHIA FILHO, A. F. C.; GARCIA, J. C. 2000. Análise e avaliação do Mercado brasileiro de sementes de milho. In: UDRY, C. V.; DUARTE, W. F. (Org.) Uma história brasileira do milho: o valor de recursos genéticos. Brasília: Paralelo 15,167-172. 3. Compania Nacional de Abastecimento – CONAB. 2007. Milho total (1ª e 2ª safra) Brasil – Série histórica de área plantada: safra 1976-77 a 2006-07. http:www.conab.gov.br/conabweb/download/safra/Milho TotalSerie Hist.xls. 4. CRUZ, I.; FIGUEIREDO, M. L. C.; OLIVEIRA, A. C.; VASCONCELOS, C. A. 1999. Damage of Spodoptera frugiperda (Smith) in different maize genotypes cultivated in soil under three levels of aluminium saturation. International Journal of Pest Management 45: 293-293. 5. Comissão Técnica Nacional de Biossegurança. CNTBio 2008. Parecer Técnico 1597/2008. Publicado no Diário Oficial da União de 14 /10/2008, Seção 1, pag. 3. 6. Comissão Técnica Nacional de Biossegurança. CNTBio 2008. Parecer Técnico 1255/2008. Publicado no Diário Oficial da União de 16 /01/2008, Seção 1, pag. 2. 7. AGBIOS, 2009. GM DataBase, (http:www.agbios.com/dbase.php?action=Submit&evidcode=NK603+x+MON810), disponível em 07/10/2009. 8. FISCHHOFF, D. A.; BOWDISH, K. S.; PERLK, F. J.; MARRONE, P. G.; MCCORMICK, S. M.; NIEDERMEYER, J. G.; DEAN, D. A.; KUSANO-KRETZMER, K.; MAYER, E. J.; ROCHESTER, D. E.; ROGERS, S. G.; FRALEY, R. T. Insect tolerant transgenic tomato plants. Biotechnology, v. 5, p. 807-813, 1987. 9. FORLANI, G.; OBOJSKA, A. BERLICKI, T.; KAFARSKI, P. 2006. Phosphinothricin analogues as inhibitors of plant glutamine synthetase. J. Agric. Food Chem. 54: 796-802. 10. Haslam, E. 1993. Shikimic acid: metabolism and metabolites. University of Sheffield, UK. 11. Steinrücken, H. C.; Amrhein, N. 1980. The herbicide glyphosate is a potent inhibitor of 5 enilpyruvyul – shikimic acid-3-phosphatase. Biochem Biophys Res Commun 94: 1207-1212. 12. OKUNUKI, H.; TESHIMA, R.; SHIGETA, T.; SAKUSHIMA, J.; AKIYAMA, H.; GODA, Y.; TOYODA, M.; SAWADA, J. Increased digestibility of two products in genetically modified food (CP4-EPSPS and Cry1Ab) after preheating. J. Food Hygienic Soc. Japan. v. 43, p. 68-73, 2002. 13. WOLFERSBERG, M. G. V-ATPASE-ENERGIZED EPITHELIA AND BIOLOGICAL INSECT CONTROL. J. EXP. BIOL. 172, 377-386, 1992. 14. Wieczorek, H.; Brown, D.; Grinstein, S.; Ehrenfeld, J. and Harvey, W. R. (1999). Animal plasma membrane energization by proton-motive V-ATPaes. Bioessays 21, 637-648. 15. Griffitts and Aroian, 2005 J. Griffitts and R. Aroian, Many roads to resistance: how invertebrates adapt to Bt toxins, BioEssays 27 (2005), pp. 614-624. 16. Shimada, N.; Miyamoto, K.; Kanda, K.; Murata, H.; 2006a. Bacillus thuringiensis insecticidal Cry1Ab toxin does not affect the membrane integrity of the mammalian intestinal epithelial cells: an in vitro study. In vitro Cellular and Developmental Biology – Animal, 42: 45-49. 17. Shimada, N.; Miyamoto, K.; Kanda, K.; Murata, H.; 2006a. Binding of Cry1Ab toxin, a Bacillus thuringiensis insecticidal toxin, to proteins of the bovine intestinal epithelial cell: an in vitro study. Applied Entomology and Zoology, 41: 295-301. 18. Stumpff, F.; Bondzio, Einspanier, A.; R. and Martens, H. Effects of the Bacillus thuringiensis toxin Cry1Ab on membrane currents of isolated cells of the ruminal epithelium J, Membr. Biol. 219 (1-3) (2007), pp. 37-47. 19. Bondzio, A,; Stumpff, F.; Scion, J.; Martens, H.; Einspanier, R. 2008. Impact of Bacillus thuringiensis toxin Cry1Ab on rumen epithelial cells (REC) – A new in vitro model for safety assessment of recombinant food compounds. Food and Chemical toxicology, 46: 1976-1984. 20. Groten, J. P.; Schöen,; E. D.; Kuper, C. F.; Van Bladeren, P. J.; Van Zorge, J. A. and Feron, V. J. Subacute of a mixture of nine chemicals in rats: detecting interactive effects with a two level factorial design. Fundamental and Applied Toxicology (1997). 21. Jonker, D.; Woutersen, R. A. and Feron, V. J. Toxicology of mixture of nephrotoxicants with similar or dissimilar mode of action. Food and Chemical Toxicology 31 (1996), pp. 1075-1082. 22. Jonker, D.; Woutersen, R. A.; Van Bladeren, P. J.; Til, H. P. and Feron, V. J. 4-week oral toxicity study of a combination of eight chemicals in rats: comparison with the toxicity of the individual compounds. Food and Chemical Toxicology 28 (1990), pp. 623-631. 23. Jonker, D.; Woutersen, R. A.; Van Bladeren, P. J.; Til, H. P. and Feron, V. J. Subacute (4-wk) oral toxicity of a combination of four nephrotoxins in rats: comparison with the toxicity of the individual compounds. Food and Chemical Toxicology 31 (1993), pp.125-136. 24. TAYLOR, M. L.; HARTNELL, G.; NEMETH, M.; KARUNANANDAA, K.; GEORGE, B. 2005. Comparison of broiler performance when fed diets containing corn grain with insect-protected (corn rootworm and European corn borer) and herbicide-tolerant (glyphosate) traits, control corn, or commercial reference corn-revisited. Poult. 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Expression of the full-length and 3’-spliced Cry1Ab gene in 135-kDa crystal protein minus derivative of Bacillus thuringiensis subsp. Cur. Microbial. 45: 133-138. 30. AQUILA, J. M.; VILLELLA, F. M. F.; FOSTER, J. E. 2002. Resistencia do milho (Zea mays L.) transgenic (Bt) à lagarta-do-cartucho, Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae). Revista Brasileira de Milho e Sorgo 1(3): 1-11. 31. WATSON, S. A.; RAMSTAD, P. E. 1987. Corn: chemistry and technology. St. Paul.: American Association of Cereal Chemist, 605p. 32. CARPENTER, J.; FELSOT, ª; GOODE, T.; HAMMING, M.; ONSTAD, D.; SANKULA, S. 2002. Comparative environmental impacts of biotechnology-derived and traditional soybean, corn, and cotton crops (CAST: 1-189). Ames, IA: Council for Agricultural Science and Technology. 33. WATSON, S. A.; RAMSTAD, P. E. 1987. Corn: chemistry and technology. St. Paul.: American Association of Cereal Chemist, 605p. 34. CANTWELL, G. E.; LEHNERT, T.; FOWLER, J. 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Genetically modified maize: pollen movement and crop co-existence. Dorchester, UK: PG Economics, 20 pp. (www.pgeconomics.co.uk/Maizepollennov2004final.pdf); 40. MESSEGUER, J.; PEÑAS, G,; BALLESTER, J.; BAS, M.; SERRA, J.; SALVIA, J.; PALAUDELMAS, M.; MELÉ, E. 2006. Pollen-mediated gene flow in maize in real situations of coexistence. Plant Biotechnology Journal. 4: 633-645. 41. NIELSEN, K. M.; BONES, A. M.; SMALLA, K.; VAN, ELSAS, J. D. 1998 Horizontal gene transfer from transgenic plants to terrestrial bacteria – a rare event? FEMS Microbiology Reviews 22, 79-103. 42. SIQUEIRA, J. O.; TRANNIN, I. C. B.; RAMALHO, M. A. P.; FONTES, E. M. G. 2004. Interferências nos agrossistemas e riscos ambientais de culturas transgênicas tolerantes a herbicidas e protegidas contra insetos. Cadernos de Ciências e Tecnologia 21: 11-81. 43. CARPENTER, J.; FELSOT, ª; GOODE, T.; HAMMING, M.; ONSTAD, D.; SANKUSA, S. 2002. Comparative environmental impacts of biotechnology-derived and traditional soybean, corn, and cotton crops (CAST: 1-189). Ames, IA: Council for Agricultural Science and Technology. 44. EDGE, J. M.; BENEDICT, J. H.; CARROLL, J. P.; REDING, H. K. 2001. Bollgard cotton: an assessment of global economic, environmental and social benefits. J. Cotton Sci 5: 121-136. 45. GIANESSI, L.; SILVERS, C.; SANKUSA, S.; CARPENTER, J. A. 2002. Plant biotechnology: current and potential impact for improving pest management in U.S. agriculture – an analysis of 40 case studies (executive summary). Washington, DC: National Center for food and Agricutural Policy. http:www.ncfap.org/40CaseStudies/NCFAB%20Exec%20Sum.pdf. 46. HUANG, J.; ROZELLE, S.; PRAY, C.; WANG, Q. 2002. Plant biotechnology in China. Science 295: 674-676. 47. XIA, J. Y.; CUI, J. J.; MA, L. H.; DONG, S. X.; CUI, X. F. 1999. The role of transgenic Bt cotton in integrated insect pest management. Acta Gossypii Sim 11: 57-64. 48. 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Molecular traditional methods

Based on the risk assessment, it can be concluded that the event shows the same risks as its conventional counterpart. Therefore the National Technical Biosafety Committee for GMO use exclusively in Health and human consumption (CTNSalud) recommends its authorization

Indonesian National Agency of Drug and Food certified food safety for GM Maize event BT11 (resistance to Lepidoptera and tolerance to Glufosinate) x GA21 (tolerance glyphosate)

Competent National Authority: Ministry of Agriculture-Jehad, Agricultural Research, Education and Extension Organization (AREEO). Risk Assessment file is uploaded. https://bch.cbd.int/en/database/RA/BCH-RA-IR-114170/2

Please see the link below.

Authorization by COFEPRIS: 51

The genetically modified SYN-BTØ11-1xMON-ØØØ21-9 maize, as described in the application, expresses the Cry1Ab protein which confers protection against certain lepidopteran pests, the mEPSPS protein which confers tolerance to glyphosate herbicides and a PAT protein which confers tolerance to glufosinate-ammonium herbicides.

UI OECD: SYN-BTØ11-1xMON-ØØØ21-9 During the risk assessment of this GMO based on existing knowledge to date, no toxic or allergic effects neither substantial nutritional changes are observed. The event is as safe as its conventional counterpart. For more detail please find attached the risk assessment summary in this page.

Backcrossing was used to move the trait into an inbred background to generate a fixed inbred for each trait . The fixed transgenic inbreds (corn Bt 11 and corn GA21) are then crossed to produce a commercial hybrid containing both events, Bt 11 x GA21.

Syngenta Philippines, Inc have filed an application with attached technical dossiers to the Bureau of Plant Industry (BPI) for a biosafety notification for direct use as food, feed and for processing under Department of Agriculture (DA)- Administrative Order (AO) No. 8 Part 5 for stacked trait product corn: Bt11 x GA21, which was developed by conventionally crossing two genetically modified corn events (Bt11 and GA21) for insect resistance and herbicide tolerance. A safety assessment of combined trait product corn: Bt11 x GA21 was conducted as per Administrative Order No. 8 Series of 2002. The focus of risk assessment is the gene interactions between the two transgenes. Review of results of evaluation by the BPI Biotech Core Team in consultation with DA-Biotechnology Advisory Team (DA-BAT) completed the approval process.

Please see the link below(in Korean).

The GM maize BT11xGA21 has been assessed in terms of the Genetically Modified Organisms Act, 1997 by the Advisory Committee, a scientific panel and the Executive Council an intergovernmental decision making body. The assessment considered amongst others the following: The source of the gene, nature of host organism, protein expression, toxicology and allergenicity issues.

Commodity : Corn / Maize (Zea mays L. )

The stacked event Bt11 x GA21 maize obtained from conventional breeding of genetically modified maize event Bt11 and event GA21 to expresses Bt-toxin (Cry1Ab protein) which provide protection to European corn borer (ECB), enzyme phosphinothricin N-acetyl transferase (PAT) which confers tolerance to glufosinate-ammonium herbicide, and mEPSPS protein which confers tolerance to glyphosate herbicide.
Maize Bt11 was generated by using PAT protein as a selectable marker enabling identification of transformed plant cells as well as a source of resistance to the herbicide known as glufosinate ammonium.

Application for food safety assessment.

The food safety assessment performed by the National Center for Genetic Engineering and Biotechnology (BIOTEC) as advisory and technical arm of Thai FDA. BIOTEC conduct food safety assessment according to codex guideline and based on the safety data and information provided by the applicant (as specified in Annex 2 attached to Notification of the Ministry of Public Health No.431). According to the existing scientific data and information available during the safety assessment, it is concluded that the stacked event Bt11 x GA21 maize is substantially equivalent to its conventional counterpart in terms of morphology, nutrition, toxicity and allergenicity.

Commodity:Corn / Maize (Zea mays L.)

 

The stacked event Bt11 x GA21 maize obtained from conventional breeding of genetically modified maize event Bt11 and event GA21 to expresses Bt-toxin (Cry1Ab protein) which provide protection to European corn borer (ECB), enzyme phosphinothricin N-acetyl transferase (PAT) which confers tolerance to glufosinate-ammonium herbicide, and mEPSPS protein which confers tolerance to glyphosate herbicide.
Maize Bt11 was generated by using PAT protein as a selectable marker enabling identification of transformed plant cells as well as a source of resistance to the herbicide known as glufosinate ammonium.

Application for food safety assessment.

 

The food safety assessment performed by the National Center for Genetic Engineering and Biotechnology (BIOTEC) as advisory and technical arm of Thai FDA. BIOTEC conduct food safety assessment according to codex guideline and based on the safety data and information provided by the applicant (as specified in Annex 2 attached to Notification of the Ministry of Public Health No.431). According to the existing scientific data and information available during the safety assessment, it is concluded that the stacked event Bt11 x GA21 maize is substantially equivalent to its conventional counterpart in terms of morphology, nutrition, toxicity and allergenicity.

Application for direct use as feed

Turkish Biosafety Law, entered in force in 2010, diverges from EU legislations in some points
 such as food and feed use require different separate applications, risk assessments and approvals.
  Addition, our Law forsees prision sentences in some circumtances of Law violation and joint
 reponsibilities for the violation. Therefore, GM product owners avoid to make application for approval
and non product developer have made application till now. Instead, some Turkish assosiations
 such as poultry producers assosiations, animal feed assosiations have applied to get approval
for import of GM products for their members. Thus, name of product applicants are not product
developers for our country.

Turkish Feed Manufacturer's Association
Turkish Poultry Meat Producers and Breeders Association
Turkish Egg Producers Association

After the evaluation of reports released by Scientific Risk Assessment Committee and Socio- economic Assessment Committee and also by considering public opinion, Biosafety Board has approved the use of genetically modified maize Bt11xGA21 and products thereof for animal feed.

Please refer to uploaded document