Food safety and quality
| share
 

OECD Unique Identifier details

MON-ØØ531-6xMON-Ø1445-2
Commodity: Cotton
Traits: Glyphosate tolerance,Kanamycin resistance,Lepidoptera resistance
Brazil
Name of product applicant: Monsanto do Brasil Ltda.
Summary of application:
Commercial Release of Genetically Modified Cotton Styled MON 531 x MON 1445
Upload:
Date of authorization: 15/10/2009
Scope of authorization: Food and feed
Links to the information on the same product in other databases maintained by relevant international organizations, as appropriate. (We recommend providing links to only those databases to which your country has officially contributed.): Center for Environmental Risk Assessment
Summary of the safety assessment:
The insect resistant and glyphosate tolerant MON 531 x MON 1445 (MON 531 x MON 1445 Cotton) was generated by crossing MON 531 cotton with MON 1445 cotton through classic genetic improvement and expresses proteins Cry1Ac (MON 531 cotton) and CP4 EPSPS (MON 1445 cotton). MON 531 cotton results from a genetic modification of a conventional variety of Coker 312 corn through a methodology mediated by Agrobacterium tumefaciens. Genes inserted in MON 531 were cry1Ac (conferring resistance to insects), nptIl (operating as a selection marker of transformed plants) and aad (operating as a selection marker of transformed bacteria in the process of cloning the plasmid of interest. The expression of cry1Ac confers resistance to some species of lepidopteran target-insects (cotton leafworm [Alabama argillacea], apple budworm [Heliothis virescens], corn earworm [Helicoverpa Zea] and pink bollworm [Pectinophora gossypiella]). Gene cry1Ac was isolated from bacterium Bacillus thuringiensis subspecies kurstaki and is commonly found in soils. MON 1445 cotton was generated from a genetic transformation of a commercial variety, Coker 312, through a method mediated by Agrobacterium tumefaciens. The transformation inserted genes cp4 epsps, nptIl, and aad to the genome of cotton variety Coker 312. The lineage resulting from the transformation expresses enzyme CP4 EPSPS coming from Agrobacterium tumefaciens sp. strain CP4, which is naturally tolerant to glyphosate. The lineage expresses also protein NPTII (Neomycin phosphotransferase II) that confers resistance to the antibiotic kanamycin. The third gene introduced, aad, codifies protein AAD, but is not expressed in plant tissues. Protein Cry1ac produced in MON 531 x MON 1445 cotton is a selective toxicity protein for some species of lepidopteran and its action is mediated by specific receptors. The protein bonds to such receptors located in the midgut of susceptible insects. Mammals, including humans, as well as fish, birds and non-target insects are devoid of such receptors. Protein CP4 EPSPS expressed in glyphosate tolerant genetically modified plants is functionally identical to EPSPS that is endogenous to plants, except for the fact that CP4 EPSPS has reduced affinity to glyphosate. In conventional plants, glyphosate bonds to the EPSPS enzyme and blocks the biosynthesis of aromatic amino acids and secondary metabolites. In glyphosate tolerant genetically modified plants, such as MON 531 x MON 1445 cotton, aromatic amino acids and other metabolites needed in the plants’ development keep being produced by the activity of CP4 EPSPS. The mode of action and biologic activity of proteins CP4 EPSPS and Cry1Ac expressed in MON 531 x MON 1445 cotton are not significantly different and do not have known interaction mechanisms that could bring adverse effects to human and animal health and to the environment. The proteins are expressed in low levels and accumulate in different cell compartments of MON 531 x MON 1445 cotton. Both such proteins have a long history of safe use translated into the consumption of cotton carrying isolated events MON 1445 and MON 531 since 1995, as well as through the mix of such events and combination of the individual events by classical genetic improvement of MON 531 x MON 1445 cotton. These proteins showed that they are easily digested in in vitro simulated digestion essays. Besides, acute oral toxicity tests with purified proteins CP4 EPSPS and Cry1Ac and subchronic toxicity tests in which such proteins were administered in magnitudes substantially above the doses found in the normal consumption of cotton showed that these proteins have no adverse effects and, therefore, are not held as a problem of health safety for humans and animals. Considering, additionally, that proteins CP4 EPSPS and Cry1Ac failed to cause toxicity in the maximum doses tested, it is highly unlikely that an interaction shall happen between such proteins in the normal doses found in food that may cause additive or synergistic effects. Studies conducted with MON 531 x MON 1445 cotton in the fields, nursery and laboratory showed that this genetic transformation event is comparable to conventional corn in what regards its reproductive, agronomic, food safety, environmental and nutritional characteristics. Quality and composition analyses of cotton kernel containing the single event MON 531 showed that the properties of genetically modified cotton and its processed fractions are comparable to the properties of conventional corn. No effect in the quality of cotton fibers was noticed as an effect of the gene insertion. Except for the tolerance to target-insects during the harvest, MON 531 x MON 1445 cotton plants were equivalent in all phenotypic and agronomic characteristics when compared to the parental non-transformed lineage and to other varieties used in commercial production. Introgression of a transgene to wild cotton plants could only take place if a strong selective advantage were granted, sufficient to overcome the disadvantages caused by alleles that are genetically linked to the transgene. However, the characteristics of herbicide tolerance is known as unable to grant the receiving genotypes any adaptive advantage outside farming areas, since outside such areas the potential receiving wild genotypes are not exposed to the selective pressure of the herbicide and, therefore, an eventual pollination of such genotypes would not result in gene introgression. Assay of phenotypical and agronomic characteristics of the single event MON 1445 conducted in Brazil had results similar to those obtained in other regions of the world in commercial and experimental farming. Apart from the tolerance to glyphosate, resulting from the expression of gene cp4 epsps, event MON 1445 displays phenotypical and agronomic characteristics equivalent to the pattern of parental lineages and commercial cultivars of conventional cotton. Taking into consideration the mode of action, specificity, history of exposure, absence of similarity with allergenic and toxic proteins, fast digestion in simulated gastric and intestinal fluids, lack of acute oral toxicity in animals and analysis of the independent scientific literature, cotton MON 531 expressing proteins Cry1Ac and CP4 EPSPS demonstrates safety for human and animal consumption comparable to the safety of conventional corn and its conventional isoline. Pursuant to Annex I of Ruling Resolution nº 5, of March 12, 2008, applicant shall have a term of thirty (30) days from publication of this Technical Opinion to make amendments to its proposed post-commercial release monitoring plan. Under Article 14 of Law nº 11,105/05, CTNBio held that the request complies with the applicable rules and legislation aimed at securing the safety of the environment, agriculture, and human and animal health. TECHNICAL OPINION I. Identification of GMO Name of GMO: MON 531 x MON 1445 Cotton Applicant: Monsanto do Brasil Ltda. Species: Gossypium hirsutum L. Inserted Characteristics: Tolerance to herbicide glyphosate and resistance to insects Method of insertion: Co-culture with Agrobacterium tumefaciens Proposed use: Production of fibers for the textile industry and grain for human and animal consumption from the GMO and its derivatives II. General Information Cotton belongs to genus Gossypium, Tribe Gossypiae, Family Malvaceae, order Malvales(1,2). The genus is divided into four sub-genuses (Gossypium, Sturtia, Houzingenia and Karpas) that, in turn, are subdivided into nine sections and several sub-sections(3). Genus Gossipyum currently encompasses fifty well differentiated species, in which the American cottons are allotretrapoid while African, Asian and Australian cottons are diploid(4). Cotton is an important cultivated plant, represented by commercial species, such as Gossypium hirsutum, Gossypium barbadensis, Gossypium arboretum, and Gossypium herbaceum. Gossypium hirsutum is the main species, responsible for about 90% of total cotton fiber produced in the world, the raw material for over 40% of human clothing(5). Cotton is held as one of the most important farm products to Brazil, for its complex processing/industrialization process and the high use of manpower. Two types of cotton plants are prevalently cultivated in Brazil: conventional cotton and caterpillar resistant genetically modified cotton. These are responsible for practically all cotton produced in the country. In addition to them, there are three other cotton plants with special genetic or ecologic characteristics are cultivated: naturally colored fiber cotton, organic cotton and agroecologic cotton. The colored cotton is almost exclusively concentrated in the State of Paraíba, and the area sowed in 2007 was about 3000 hectares. Plots of agroecologic cotton were cultivated by 235 farmers in the semiarid biome of four states in the Northeastern region with an output of 42 tons(6). Chains of special, conventional and transgenic cotton plants have lived together in a satisfactory way, without any known record of coexistence problems. The area occupied by cotton plants in the 2007/2008 crop was about one million and one hundred thousand hectares, of which over 85% in the Cerrado biome, especially in the States of Mato Grosso, Bahia, Goiás and Mato Grosso do Sul. The remaining cotton farms are present in other Brazilian States, particularly in the semi-arid part of the Northeastern region and States of Paraná, Minas Gerais and São Paulo(7). Besides the herbaceous cotton, three other cotton plants grow in Brazil, all of them Allotetraploid and, therefore sexually compatible with other cultivars. None of such plants is held as a pest in agricultural and natural environments. The species G. barbadense has domestication center in the Northern Peru and Southern Ecuador(8). It was introduced by pre-Colombian peoples and its fiber was used to make textile craftsmanship by some indigenous ethnic groups before the Portuguese arrival(9). Its use as a textile plant spread among colonizers, but started its decadence with the dissemination of the two exotic races of G. hirsutum. G. barbadense is not found in natural environments and is maintained basically as a backyard plant. It is widely distributed across most of the country and the in situ conservation is directly linked to the traditional maintenance of use as a medicine plant(10). The only indigenous species in Brazil is G. mustelinium, with natural distribution restricted to the Northeastern semi-arid(4,11). Known populations are restricted to the States of Bahia and Rio Grande do Norte, in municipalities that are not producers of herbaceous cotton. Two problems affect the in situ maintenance of G. mustelinium. The first, and most severe, is the destruction of gallery forest of rivers and intermittent rivulets, the habitat of the species. The second is the extensive cattle rising of the region, especially goats. These animals feed on sprouts, leaves, fruits, seed and stalk bark, affecting the development and, in some cases, killing adult plants. Renewal of populations is also affected, since grazing on young plants causes their partial destruction(10). The distance among known populations and cotton producing regions prevents the cross of G. mustelinum with herbaceous cotton in the tilth. A third type of cotton plant is known as mocó cotton and belongs to a race different from the same species of the herbaceous cotton (G. hirsutum r. marie galante (Watt) Hutch.). Its origin is the Antilles and the history of its introduction to Brazil is uncertain, including hypotheses that it had been brought by Netherlanders or Africans during colonial period(9). Mocó cotton plant was extensively cultivated in the Northeastern semi-arid up to the end of the 1980s, when different problems abruptly interrupted its cultivation(12). The original centers of the species Gossypium hirsutum are in Mexico and Guatemala, while the ones of Gossypium barbadensis are in Peru and Bolivia(13). Allotetraploid species have in their genome a combination of genomes of two different diploid species(13). Insect resistant and glyphosate tolerant cotton MON 531 x MON 1445 (MON 531 x MON 1445 Cotton) was generated through the crossing of MON 531 cotton with MON 1445 cotton by classical genetic improvement and expresses proteins Cry1Ac (MON 531 cotton) and CP4 EPSPS (MON 1445 cotton). MON 531 cotton is the result of a genetic modification of the conventional variety Coker 312 cotton through a methodology mediated by Agrobacterium tumefaciens. MON 1445 cotton was generated by genetic transformation of the commercial variety Coker 312, through a system mediated by Agrobacterium tumefaciens. The transformation inserted genes cp4 epsps, nptIl and aad in the genome of the Coker 312 variety cotton. III. Description of the GMO and Proteins Expressed. MON 531 x MON 1445 cotton was obtained from crossing of genetically modified cotton containing the single event MON 531 with MON 1445 cotton through classical genetic improvement and expresses proteins Cry1Ac (MON 531 cotton) and CP4 EPSPS (MON 1445 cotton). MON 1445 cotton was genetically modified from the transformation of the commercial variety Coker 312 with plasmid PVGHGT07, by a system mediated by Agrobacterium tumefaciens. The transformation inserted genes cp4 epsps, nptII, gox and aad in the genome of this cotton variety(14). The lineage resulting from the transformation expresses enzyme CP4 EPSPS. The lineage also expresses protein NPTII, which grants resistance to aminoglycosylated antibiotics, enabling selection of cells transformed withy gene cp4 epsps in a culture medium containing the antibiotic kanamycin in the in vitro phases of the transformation process. The third gene aad introduced codifies protein AAD. The gene is not expressed in plant tissues for being under control of a prokaryotic promoter. MON 531 cotton results from genetic modification of conventional cotton variety Coker 312 through a methodology mediated by Agrobacterium tumefaciens using vector PV-GHBKW. Genes inserted in MON 531cotton were cry1Ac, granting resistance to insects, nptII and aad. The agronomically relevant gene in MON 531 cotton is cry1Ac that grants the characteristic of resistance to certain target-insects of the Lepidoptera Order (cotton leafworm [Alabama argillacea], tobacco budworm [Heliothis virescens], corn earworm [Helicoverpa Zea] and pink bollworm [Pectinophora gossypiella]). Gene cry1Ac was isolated from Bacillus thuringiensis subspecies kurstaki and its coding region was constructed by combination of the first 1,398 nucleotides of gene cry1Ab(15) with nucleotides numbered 1,399 to 3,534 of gene cry1Ac. Enzymatic hydrolysis from the whole protein to the active protein takes place in the presence of enzymes, both in vivo and in vitro. Protein Cry1Ac, produced in MON 531 x MON 1445 cotton, has an active nucleus with insecticide action that is resistant to trypsin, with the size of about 600 amino acids. Both regions of the modified gene cry1Ac were genetically improved to increase expression in plants. This way, the plants tested showed a significant increase in producing protein Cry1Ac(16,17). Despite modifications, the whole gene cry1Ac introduced in vector PV-GHBK04 during the genetic transformation codifies a protein that is 99.4% identical to the protein Cry1Ac found in nature(18) and features the same insecticide specificity(19). Protein Cry1Ac was characterized by physical and functional analyses including: determination of molecular weight; sequencing of N-terminal amino acids; immunoreactivity; absence of glycosylation; and insecticide activity. In all parameters assayed, protein Cry1Ac purified from MON 531 x MON 1445 cotton was equivalent to the protein produced and purified from Escherichia coli. Protein Cry1Ac produced in MON 531 x MON 1445 cotton is a selectively toxic protein to some lepidopteran insects(20, 21, 22, 23, 24) and its action is mediated by specific receptors. Protein Cry1Ac bonds to these receptors located in the midgut of susceptible insects. Anyway, mammals, including humans, are devoid of such receptors, the same as fish, birds and non-target insects. Understanding the mode of action of Cry proteins is a prerequisite for grasping the specificity of such proteins. They are produced in general as high molecular weight pro-toxins. When ingested by the target pest, proteases similar to trypsin or chymotrypsin that are present in the insect’s midgut break protein Cry, generating an active tryptic nucleus with lower molecular weight(25). The active toxins structure(26, 27) is defined by three domains (I, II and III), and protein Cry1Ac possesses such domains. Domain I is responsible by pore formation and domain II by specificity. Domain III is involved both in bonding to the specific receptor and pore formation. The domains define, therefore, the bonding with the receptor and activity of the ionic channel(28). Pores formed by the ionic channel activity result in an osmotic unbalance followed by cell lysis(29). Specificity of Cry proteins for some species of insects is a consequence of the bonding with the receptor. Studies on the mode of action at molecular level indicate that protein Cry1Ac bonds to receptors made of aminopeptidase N and of a protein of 210 kD, similar to cadherin(30, 31, 32). Domain II of protein Cry1Ac grants a large part of the toxicity to lepidopterans, promoting a correct bonding with receptors(33). Studies showed that the surface of intestinal cells in mammals fail to have receptors to proteins Cry of Bacillus thuringiensis, hence humans are not susceptible to proteins Cry(34, 35, 36). Gene cp4 epsps codifies protein CP4 EPSPS that is responsible by the tolerance displayed by MON 531 x MON 1445 cotton to the glyphosate herbicide. The coding region of gene cp4 epsps derives from soil Agrobacterium tumefaciens sp., strain CP4. The bacterium was identified in a screening of microorganisms resistant to the glyphosate molecule action(37, 38). Agrobacterium tumefaciens sp. strain CP4, as well as other soil bacteria and some soil fungi, are resistant to the action of glyphosate because they possess enzyme EPSPS that is little sensitive to the action of this herbicide(39). Protein CP4 EPSPS expressed in glyphosate tolerant genetically modified plants is an enzyme functionally identical to the endogenous EPSPS of conventional plants, except that protein CP4 EPSPS has reduced affinity for glyphosate(40). In conventional plants, glyphosate bonds to enzyme EPSPS and checks biosynthesis of 5-hydroxyl shikimate-3-phosphate, hindering formation of aromatic amino acids and secondary metabolites(41, 42), while in genetically modified glyphosate tolerant plants, aromatic amino acids and metabolites needed for the development of plants keep being produced by the CP4 EPSPS activity(42, 37). Comparing kinetic parameters of proteins CP4 EPSPS and endogenous EPSPS helped elucidate the mechanism of glyphosate. The mechanism operates by inhibiting the activity of the endogenous EPSPS through the formation of a complex EPSPS shikimate-3-phosphate(S3P)-glyphosate, which takes place after the bonding of EPSPS-S3P. The glyphosate bond failed to show competitiveness with S3P, though it is competitive with phosphoenolpyruvate (PEP). Therefore, protein CP4 EPSPS is highly tolerant to glyphosate and strong bonding with the phosphoenolpyruvate (PEP) substrate. A number of bacterial EPSPS proteins possessing tolerance to glyphosate were described(39). The CP4 EPSPS amino acid sequence was aligned with consensus sequences of other EPSPS previously known. Several identified residues with important functions in EPSPS proteins are preserved in CP4 EPSPS. Homology identification between CP4 EPSPS and other EPSPS proteins in critical catalytic sites show the relation among such enzymes. Three-dimensional crystallography studies revealed that the CP4 EPSPS structure exhibits the same folding model of EPSPS found in Escherichia coli. Thermal stability of EPSPS activity was assayed to determine whether such activity may be inactivated in glyphosate tolerant genetically modified cultures. Incubation of protein CP4 EPSPS at 55ºC for fifteen minutes caused a reduction of 50% in the activity measured at a 25ºC incubation, while enzymatic activity was completely eliminated after fifteen minutes of incubation at 65ºC. Another aspect relates to determination of enzymatic activity in acid environments, such as the human stomach, in case such enzymatic activity were maintained after the processing. Protein CP4 EPSPS pH dependency was measured from pH 4,0 to pH 11.0. The results lead to the conclusion that the maximum enzymatic activity took place between pH 9.0 and pH 9.5. There was no enzymatic activity detected at pH 5.0. The results established that CP4 EPSPS has no enzymatic activity in acid environments of the human stomach and that protein CP4 EPSPS expressed in MON 531 x MON 1445 cotton is the same protein expressed in other glyphosate tolerant cultures, such as soybeans, maize and canola. The mode of action and biologic activities of proteins CP4 EPSPS and Cry1Ac expressed in MON 531 x MON 1445 cotton are significantly different. Both proteins, as well as their action mode, fail to have known interaction mechanisms that could cause adverse effects to human and animal health and to the environment. The proteins accumulate in different cell compartments of MON 531 x MON 1445 cotton. EPSPS proteins are directed to the chloroplasts of plants at the specific site of metabolic action is found. Several plastid proteins are coded by nuclear genes and synthesized as precursor molecules with high molecular weight. The additional weight of such precursor proteins is due to the presence of extension N-terminal, the so-called transit peptide. The transit peptide is necessary and sufficient to carry the proteins through the cytoplasm to the cell chloroplast(44,45,46,47). Protein EPSPS produced in MON 1445 cotton and MON 531 x MON 1445 cotton is directed to the chloroplast, the site of action of all EPSPS proteins(48). Proteins designed to act in the cell cytoplasm do not need any type of N-terminal peptide. Since protein Cry1Ac expressed in MON 531 and MON 531 x MON 1445 cotton causes insect control through an insecticide action in the gastrointestinal tract of target lepidopteran, the use of a transit peptide to the chloroplast was unnecessary in assembling the DNA sequence used in the genetic modification. Action of Cry1Ac protein is only effective in case the target insect feeds on MON 531 of MON 531 x MON 1445 cotton. Summarizing, proteins CP4 EPSPS and Cry1Ac present in MON 531 x MON 1445 cotton are accumulated in different cell compartments and have separate and non-interactive metabolic functions. This way, protein CP4 EPSPS is directed to the chloroplast while protein Cry1Ac is accumulated in the cytoplasm. Proteins CP4 EPSPS and Cry1Ac are expressed at low levels in single cotton events MON 531 and MON 1445. With this expression at low levels, there is limited likelihood of biochemical interaction between proteins CP4 EPSPS and Cry1Ac in the complex matrix of a plant, not to mention that the proteins are accumulated in different plant sites. Therefore, the likelihood that such proteins interact between them is minimal, a fact microscopically ratified through analyzing agronomic and phenotypic characteristics related to efficacy and selectivity of MON 531 x MON 1445 cotton in the fields. The expected level of tolerance to glyphosate in MON 531 x MON 1445 cotton, as a result of the continuous action of CP4 EPSPS protein in the presence of the glyphosate herbicide, is the same as in cotton containing the event isolated from MON 1445 cotton, while the expected level of controlling target insects, due to the efficacy of control promoted by protein Cry1Ac is – the same way – equal to that of cotton containing the single event MON 531. Because proteins CP4 EPSPS and Cry1Ac are expressed in low concentrations in MON 531 x MON 1445 cotton, a potential exposure to these proteins is extremely low in human and animal food. The proteins showed to be rapidly digested in in vitro simulated digestion essays. Besides, acute oral toxicity tests with purified proteins CP4 EPSPS and Cry1Ac, and sub-chronic toxicity tests where such proteins were administered in doses substantially above the ones found in normal consumption of cotton, showed that the proteins failed to have any adverse effects and, therefore, are not a food safety issue for humans and animals. Given that proteins CP4 EPSPS and Cry1Ac have no toxicity at the top doses tested – and taking into consideration that enzyme CP4 EPSPS has high affinity with the substrate, besides failing protein Cry1Ac to display enzymatic activity – it is highly unlikely that an interaction takes place between such proteins in the normal doses found in food in a way to cause additive or synergistic effects. A number of reports in the scientific literature showed that such interactions are inexistent, especially when such substances are administered in doses substantially below the levels of non-observed effects (NOEL)(49, 50, 51, 52). Gene nptII codifies protein Neomycin Phosphotransferase II, granting tolerance to antibiotics neomycin and kanamycin. Gene aad codifies protein AAD (3”(9) O aminoglycoside adeniltransferase – marker of resistance selection to antibiotics). Gene gox codifies enzyme GOX (glyphosate oxyredutase), responsible for metabolizing the glyphosate herbicide. The coding region of gene gox derives from bacterium Ochrobactrum anthropi, strain LBAA, isolated in sludge discarding and treatment reservoirs for effluents of industrial glyphosate production(53, 54). Enzyme GOX has the ability to degrade glyphosate into aminomethylphosphonic acid (AMPA) and glyoxylate. Conversion of glyphosate into aminomethylphosphonic acid is the main degradation way of glyphosate in soil(55, 56, 57). Despite being present in plasmid PV-GHGT07, gene gox was not transferred to cotton and hence protein GOX failed to be detected in MON 531 x MON 1445 cotton(58). IV. Aspects Related to Human and Animal Health Protein CP4 EPSPS is an enzyme present in all plants and in a large number of microorganisms(46), while protein Cry1Ac has no enzymatic activity in plants, therefore fails to affect the plant’s metabolism. Safety of Cry1Ac and EPSPS proteins was duly assayed by CTNBio(59, 60). Due to the rigorous specificity for the substrates, enzymes EPSPS bond only S3P, PEP and glyphosate. The only known resulting metabolic product is 5 enolpyruvylshikimate-3-phosphate acid, the penultimate step of the shikimic acid pathway. Shikimic acid is a precursor for the biosynthesis of aromatic amino acids (phenylalanine, tyrosine and tryptophan) and several secondary metabolites, such as tetrahydrofolate, ubiquinone and K vitamin(61). Though the shikimic acid (or shikimate) pathway and proteins EPSPS do not occur in mammals, fish, birds, reptiles and insects, they are very important to plants. It is reckoned that aromatic molecules, all derived from shikimic acid, represent no less than 30% of the plant’s dry weight(62, 63). Studies were conducted using simulated gastric (pH 1.2) and intestinal (pH 7.5) juices, The degradation rate of protein CP4 EPSPS (mature protein, without the transit peptide) was assayed through Western blot analyses, where it was shown that protein CP4 EPSPS and peptides degrade in less than fifteen seconds after being exposed to gastric juice. In simulated intestinal juice, degradation of protein CP4 EPSPS took place in a period shorter than ten minutes(64). Degradation rate of protein NPTII was assayed through enzymatic activity. The result showed that the protein was destroyed after two minutes of incubation in simulated gastric juice and fifteen minutes of incubation in simulated intestinal juice(65). Based on these results, the forecast is that the new proteins CP4 EPSPS and NPTII expressed in MON 531 x MON 1445 cotton are rapidly digested in the digestive tract of mammals. Exogenous proteins rapidly digested pose minimum toxicity or allergic risk when compared to other safe dietary proteins(66, 67). Effects of acute oral toxicity of proteins CP4 EPSPS, Cry1Ac and NPTII were conducted in mice(68, 65) and the results demonstrated no evidence of toxicity when protein CP4 EPSPS was administered orally by gavage to mice, the same for proteins Cry1Ac and NPTII. Especially, no abnormal clinical signs were observed in any animal during the study, showing that there was no significant statistic difference in corporal weight, cumulative corporal weight or food consumption between treatment and control groups. All animals were sacrificed at the seventh post-dosage day and submitted to necropsy, where no pathological effect was noticed(69). Based on data of CP4 EPSPS protein expressed in the kernel of cotton containing the single event MON 1445, the dose of 572 mg/kg of body weight is equivalent to a consumption from 3.2 to 7.1 kg of kernel for kg of corporal weight. In the study conducted with NPTII protein, doses of 5000, 1000 and 100 mg of NPTII per kg of body weight were administered orally through gavage to mice. It shall be stressed that the dose corresponding to 5000 mg per kg of body weight is equivalent to approximately 700 kg of kernel per kg of body weight. Completed the experiments, the results demonstrated an absence of pathological effects(65). The outcomes of these studies confirmed that proteins CP4 EPSPS and NPTII fail to possess acute oral toxicity for mammals. The results are in line with the absence of any similarity with the amino acid sequence with known toxic proteins and the rapid digestion of such proteins in gastric and intestinal juices, as mentioned in this document. IV. Environmental and Agronomic Aspects Phenotype and agronomic assays of MON 531 x MON 1445 cotton, as well as assays on efficacy and tolerance to glyphosate were conducted in Brazil. The assays were field studies held in typical location for the culture of cotton, simulating the Brazilian commercial production. The studies were conducted during the 1999/2000 crop in three places (Ituverava, State São Paulo; Rondonópolis, State of Mato Grosso; and Capinópolis, State of Minas Gerais. The experiment followed conventional agronomic practices, typically used in assessment of new cultivars and included the following parameters: number of nodes, height, first branch position, bolls per linear meter, susceptibility to diseases (Colletotrichium sp., Romularia sp., Blue Disease, Cotton Anthocyanosis and Alternaria sp.), open bolls per meter, and productivity. No significant differences were reported in both: phenotypic and morphological aspects; and agronomic performance of DP50 (conventional) and DP50R (MON 531 x MON 1445) for the parameters assayed. During the 2002/2003 crop phenotypic, morphological and agronomic parameters were again assayed and expanded. Experiments were conducted in two locations (Santa Cruz das Palmeiras, State of São Paulo, and Santa Helena de Goiás, State of Goiás). The experiments compared the development of Roundup Ready MON 531 x MON 1445 cotton, conventional CP50 cotton and eight commercial cotton cultivars, four in each location. Regarding the remaining cultivars, all plots were equally treated with fertilizers and disease and insect controlling agrochemicals, using the same products and doses and following conventional agronomic practices, typically used in each region. Comparisons between genetically modified cotton and conventional varieties were conducted during the harvest for the following parameters: plant vigor, flowering cycle, height, maturity precociousness, cycle up to harvest, boll retention by the capsule after dehiscence, weight of open boll, average weight of thousand kernels, fiber percentage, susceptibility to diseases and pests (Colletotrichium sp., Romularia sp., Blue Disease, Cotton Anthocyanosis, and Alternaria sp.), productivity, and fiber quality. No significant differences were reported between the genetically modified cultivar, both with and without the use of glyphosate, and its parental conventional regarding assayed phenotypic, morphological and agronomic parameters. Summarizing, phenotypic and agronomic characteristic assays of MON 531 x MON 1445 conducted in Brazil showed results similar to those found in other regions of the world in experimental and commercial cultivars. Except for glyphosate tolerance and insect resistance, resulting from expression of gene cp4 epsps and Cry1Ac, MON 531 x MON 1445 Cotton displays phenotypic and agronomic characteristics that are similar to that of conventional parental lineages and/or cultivars of conventional cotton. VI. Restrictions to the use of GMO and its derivatives Technical reports related to agronomic performance concluded that there is equivalence between transgenic and conventional plants. Thus, the information indicates that transgenic plants are not fundamentally different from the genotypes of non modified cotton, excepted for the tolerance to glyphosate. Additionally, there are no records of adverse reactions to the use of Roundup Ready event 1445 Cotton. Therefore, there are no restrictions to the use of this cotton or its derivatives in human and animal food. According to Article 1 of Law no. 11,460, of March 21, 2007, “research and cultivation of genetically modified organisms are forbidden in indigenous and conservation unit areas”; as well as in Municipalities mentioned in Annex to the Ministry of Agriculture (MAPA) Directive nº 21, of 01.16.2006. VIII. Conclusion Whereas MON 531 x MON 1445 cotton (Gossypium hirsutum) belongs to a well characterized species with a steady history of safety to human consumption; and that genes cry1Ac and cp4 epsps introduced in this variety codify proteins that are ubiquitous in nature, present in plants, fungi and microorganisms included in the alimentary diet of humans and animals; whereas insertion of such genotype took place through classic genetic improvement resulting in insertion of a stable and functional copy of genes cry1Ac and cp4 epsps that granted plants tolerance to glyphosate herbicide and resistance to insects; whereas centesimal composition data fail to exhibit significant differences between genetically modified and conventional varieties, suggesting nutritional equivalence between them; whereas CTNBio assayed the events separately and issued an opinion favorable to commercial release of each event individually; Whereas: 1. Comparative biochemical studies indicate that proteins Cry1Ac and CP4 EPSPS display important difference in their mode of action and, therefore, the likelihood of interaction between these two proteins is low; 2. protein cry1Ac belongs to the Cry family of proteins, derived from Bacillus thuringiensis, an organism that has been used commercially for over forty years in producing insect control microbial formulations; 3. the history of safe use and data of multiple studies support the conclusions on safety of MON 531 x MON 1445 cotton and proteins cry1Ac and cp4 epsps; 4. agronomic and efficacy assessment of MON 531 x MON 1445 cotton indicate that the event fails to lead to any other characteristics except the expected ones; 5. phenotypic, agronomic and ecological interactions assays indicate that MON 531 x MON 1445 cotton is comparable to conventional cotton and fails to pose higher risk to become an invading plant; 6. protein ep4 epsps is rapidly degraded in the gastrointestinal tract of humans and animals; and, in addition, because of its mode of action, specificity, and absence of homology with toxic sequences, the protein does not represent risk to human health nor to the environment; 7. laboratory studies with indicating species showed that protein cry1Ac fails to cause adverse effects to non-target organisms; 8. potential allergenicity assay studies evidenced that proteins CP4 EPSPS and NPTII are not detectable in cotton products used as human and animal food and do not pose significant risk; 9. assays of phenotypic agronomic characteristics of Roundup Ready Cotton event 1445 conducted in Brazil display results that are similar to the ones found in other regions of the world in commercial and experimental crops; 10. field experiments and commercial crops with several generations and derivatives of MON 531 x MON 1445 cotton showed a stable phenotype for tolerance to the glyphosate herbicide and resistance to insects; 11. studies in rats demonstrated that the paste of cotton MON 531 x MON 1445 kernel is as safe and nutritional that the paste of conventional cotton kernel; 12. studies in quails it was demonstrated that the paste of cotton kernel is equivalent to the paste of conventional cotton kernel used as food. For the foregoing and considering internationally accepted criteria used in the process of risk analysis of genetically modified raw materials, it is possible to conclude that MON 531 x MON 1445 cotton is as safe as its conventional equivalent. CTNBio finds that the activity is not a potential cause of significant degradation of the environment or harm to human and animal health. Restrictions to the use of the GMO and its derivatives hinge on the provisions of Law nº 11,460, of March 21, 2997, and CTNBio Ruling Resolution nº 03 and Ruling Resolution nº 04. CTNBio analysis took into consideration opinions by the Commission members; ad hoc consultants; documents delivered by applicant to the CTNBio Executive Secretariat; results from planned releases into the environment; and lectures, texts and discussions of the public hearing held on 08.17.2007. Independent scientific publications of applicant, conducted by third parties, were additionally considered and consulted. Under Annex I of CTNBio Ruling Resolution no. 5, of March 12, 2008, applicant shall have a term of thirty (30) days from the publication of this Technical Opinion to adapt its proposed post-commercial release monitoring plan. VII. Bibliography 1. Fryxell, P. A. The natural history of the cotton Tribe Malvaceae (Tribe Gossypieae). Texas A&M University Press, College Station. 2. Munro, J. M. 1987. Cotton 2nd Ed. John Wiley & Sons, New York, NY. 3. Fryxell, P. A.; Craven, L. A. e Stewart, J. McD. 1992. A revision of Gossypium Sect. Grandicalyx (Malvacease) including the description of six new species. Systematic Botany, v. 17, n. 1, p. 91-114. 4. Freire, E. C. 2000. Distribuição, coleta, uso e preservação das espécies silvestres de algodão no Brasil. Embrapa, Campinas Grande. 22p. 5. Beltrão, N. E. M. 2003. Documento 117: Breve história do algodão no nordeste do Brasil. Campina Grande: Empresa Brasileira de Agropecuária, Centro Nacional de Pesquisa de Algodão, 20p. 6. LIMA, P. J. B. F. 2007. Algodões Transgênicos: grave ameaça ao algodão agroecológico e orgânico da Agricultura Familiar no Semi-árido nordestino. ESPLAR: documento apresentado em audiência pública da CNTBio sobre algodoeiros geneticamente modificados. 7. Instituto Brasileiro de Geografia e Estatística, IBGE, 2008. 8. BRUBAKER, C.; BOURLAND, E. M.; WENDEL, J. E. 1999. The origin and domestication of cotton. In: SMITH, C. W.; COTHEN, J. T. Cotton: origin, history, and production. New York: John Wiley & Sons, p. 3-31. 9. MOREIRA, J. A. N.; SANTOS, R. F. 1994. Origem, crescimento e progesso da cotonicultura do Brasil. Campinas Grande: EMBRAPA-CNPA / Brasília: EMBRAPA-SPI, 169p. 10. BARROSO, P. A. V.; FREIRE, E. C.; AMARAL, J. A. B. do; SILVA, M. T. 2005. Zonas de exclusão de algodoeiros transgênicos para preservação de espécies de Gossypium nativas ou naturalizadas. Campina Grande: Embrapa Algodão, 7p. (Comunicado Técnico, 242). 11. JOHNSTON, J. A.; MALLORY-SMITH, C.; BRUBAKER, C. L.; GANDARA, F.; ARAGÃO, F. J. L.; BARROSO, P. A. V.; QUANG, V. D.; CARVALHO, L. P. DE; KAGEYAMA, P.; CIAMPI, A. Y.; FUZATTO, M.; CIRINO, V.; FREIRE, E. 2006. Assessing gene flow from Bt cotton in Brazil and its possible consequences. 2006. In: HILBECK, A.; ANDOW, D.; FONTES, E. M. G. Environmental risk assessment of genetically modified organism: methodologies for assessing Bt cotton in Brazil. p. 261-299. 12. BELTRÃO, N. E. de M. 1999. Algodão brasileiro em relação ao mundo: situação e perspectiva, In: BELTRÃO, N. E. de M. (Ed.). O agronegócio do algodão no Brasil. Brasília: Embrapa Comunicação para Transferência de Tecnologia, v. 1, p. 17-27. 13. Stephens, S. G. 1967. Evolution under domestication of the new world cottons (Gossypium spp.). Ciência e Cultura, São Paulo, v. 19(1): 118-134. 14. Serdy, F. S. & Nida, D. L. 1995. Petition for determination for non-regulated status, cotton with the Roundup gene, lines 1445 and 1698. Petition submitted to USDA/APHIS/BBEP on February 10, 1995. 15. Fischhoff, D. A.; K. S. Bowdish; F. J. Perlak; P. G. Marrone; S. M. McComick; J. G. Niedemeyer; D. A. Dean; K. Kusano-Kretzmer; E. J. Mayer; D. E. Rochester; S. G. Rogers. e R. T. Fraley. 1987. Insect tolerant transgenic tomato plants. Biotechnology 5: 807-813. 16. Perlak, F.; Funch, R.; Dean, D.; McPherson, S.; Fischoff, D. 1990. Modification of the coding sequences enhances plant expression of insect control protein genes. Proc. Nat’l. Acad. Sci (USA) 88: 3324-3328. 17. Perlak, F. J.; Fuchs, R. L.; Dean, D. A.; McPherson, S. L.; e Fischhoff, D. A. 1991. Modification of the coding sequence enhances plant expression of insect control protein genes. Proc. Nat’l Acad Sci U S A 88: 3324-8. 18. Adang, M. J.; Staver, M. J.; Rocheleau, T. A.; Leighton, J.; Barker, R. F.; e Thompson, D. V. 1985. Characterized full-length and truncated plasmid clones of the crystal protein of Bacillus thuringiensis subsp. kurstaki HD-73 and their toxicity to Manduca sexta. Gene 36: 289-300. 19. Geiser, M.; Schweitzer, S. e Grimm, C. 1986. The hypevariable region in the genes coding for entomopathogenic of Bacillus thuringiensis: nucleotide sequence of the kurhdl gene of subsp. kurstaki HD1. Gene 48: 109-18. 20. Aronson, A. I.; Beckman, W. e Dunn, P.. 1986. Bacillus thuringiensis and related insect pathogens. Microbiol. Rev. 50 1:|- 24. 21. Dulmage, H. T. 1981. Insecticidal activity of isolates of Bacillus thuringiensis and their potential for pest control, p. 193-222 Microbial control of pests plant diseases 1970-1980, Vol. 11. Bunger, H. D. 22. Klausner, A. 1984. Microbial insect control. Biotechnology: 408- 419. 23. Mackintosh, S. C.; Stone, T. B.; Sims, S. R.; Hunst, P. L; Greenplante, J. T.; Marrone, P. G.; Pelak, F. J.; Fischhoff, D. A. and Funchs, R. L. 1990. Specificity and efficacy of purified Bacillus thuringiensis proteins against agronomically important insects. J. Invertebr. Pathol. 56: 258-266. 24. Whiteley, H. R. e H. E. Schnepf. 1986. The molecular biology of parasporal crystal body formation in Bacillus thuringiensis. Annu Rev Microbiol 40: 549-76. 25. Hofte, H.; e H. R. Whiteley. 1989. Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol Rev 53: 242-55. 26. Grochulski, P.; Masson, L.; Borisova, S.; Pusztai-Carey, M.; Schwartz, J. L; Broussearu, R.; e Cygler, M. 1995. Bacillus Thuringiensis CryA(a) insecticidal toxin: crystal structure and channel formation. J. Mol. Biol. 254447-464. 27. Li, J.; Carroll, J.; Ellar, D. J. 1991. Crystal structure of insecticidal d-endotoxin from Bacillus Thuringiensis at 2.5 a resolution. Nature 353: 815-821. 28. Schnepf, E.; Crickrnore, N.; Van Rie, J.; Lereclus, D.; Baurn, J.; Feitelson, J.; Zeigler, D. R. e Dean, D. H. 1998. Bacillus Thuringiensis and its pesticidal crystal proteins. Microbiol. Mol. Biol. Rev. 62: 775-806. 29. Gill, S. S.; Cowles, E. A. e Pietrantonio, P. V. 1992. The mode of action of Bacillus Thuringiensis endotoxins. Ann. Rev. Entomol. 37: 615-636. 30. Francis, B. R. e L. A. Bulla, Jr. 1997. Further characterization of BT-RI, the cadherin-like receptor for Cry1Ab toxin in tobacco hornworm (Manduca sexta) midguts. Insect Biochem MOI Biol 27: 541-50. 31. Keeton, T. P. & Bulla, L. A., Jr. 1997. Ligand specificity and affinity of BT-R1, the Bacillus Thuringiensis Cry 1A toxin receptor from Manduca sexta, expressed in mammalian and insect cell cultures. Applied and Environmental Microbiology 63, 3149-3125. 32. Masson, L.; Lu, Y. J.; Mam, A.; Brousseau, R.; Adang, M. J. 1995. The Cry1A(c) receptor purified from Manduca sexta displays Multiple specificities. J Biol Chem 270: 20309-15. 33. Rajarnohn, F.; Alcantara, E.; Lee, M. K.; Chen, X. J.; Curtiçç, A.; Dean, D. H. 1995. Single amino acid changes in domain II of Bacillus Thuringiensis Cry1Ab delta-endotoxin affect irreversible binding to Manduca sexta midgut membrane vesicles. J Bacteriol 177: 2276-82. 34. Hofmann, C.; Vanderbruggen, H.; Hoefte, H.; Van Rie, J.; Jansens, S.; Van Mellaert, H. 1988. Specificity of Bacillus Thuringiensis delta- endotoxins is correlated with the presence of high-affinity binding sites in the brush border membrane of target insect midguts. Proc. Natl. Acad. Sci. USA 85: 7844-7848. 35. Noteborn, H. P. J.M.; Bienenmann- Plaum, M. E.; Van Den Berg; Alink, G. M.; Zolla, L.; Reynaerts, A.; Pensa, M.; Kuiper, H. A. 1995. Safety assessment of the Bacillus Thuringiensis insecticidal crystal protein CRY1A (b) expressed in transgenic tomatoes. Genetically modified foods. 36. Sacchi, V. F.; Parenti, P.; Hanozet, G. M.; Giordana, B.; Luthy, P.; Wolfersberger, M. G. 1986. Bacillus Thuringiensis toxin inhibits K+ gradient dependent amino aad transport across the brush membrane of Pieris brassicae midgut cells. FEBS Letiers 204: 213-218. 37. Padgette, S. R.; Kolacz, K. H.; Delannay, X.; Re, D. B.; LaVallee, B. J.; TInius, C. N.; Rhodes, W. K.; Otero, Y. I.; Barry, G. F.; Eichholtz, D. A.; Peschke, V. M.; Nida, D. L.; Taylor, N. B. e Kishore, G. M. 1996b. Development, identification and characterization of a glyphosate-tolerant soybean line. Crop Science. 35: 1451-1461. 38. Padgette, S. R.; Re, D. B. e Barry, D. E. 1995. New weed control opportunities: development of glyphosate tolerant soybean. In: Duke, S. O. Ed. Herbicide resistant crops. CRC Press, Boca Raton. 39. Schulz, A.; Kruper, A. and Amrhein, N. 1985. Differential sensitivity of bacterial 5-enopyruvyl-shikimate-3-phosphate synthases to the herbicide glyphosate. FEMS Microbiol. Lett. 28: 297-301. 40. Padgette, S. R.; Barry, G. F.; Re, D. B.; Eichholtz, D. A.; Weldon, M.; Kolau, K.; Kishore, G. M. 1993. Purification, cloning and e characterization of a highly glyphosate-tolerant 5-enolpyruvylshikimate-3-phosphate synthase from Agrobacterium sp. Strain CP4. Monsato Technical Report MSL 12738. 41. Haslam, E. 1993. Shikimic acid: metabolism and metabolites. University of Sheffield, UK. 42. Steinrücken, H. C.; Arnrhein, N. 1980. The herbicide glyphosate is a potent inhibitor of Senolpynivyl - shikimic acid-3-phosphate synthase. Biochem Biophys Res Comrnun 941207-1212. 43. Padgette, S.; Taylor, N.; Nider, D. e. al. 1996a. The composition of glyphosate-tolerant soybean seed is equivalent to that of conventional soybeans. J Nutr 126: 702-716. 44. Chua, N. H.; Schrnidt, G. W. 1978. Post-translational transport into intact chloroplasts of a precursor to the small subunit of ribulose-13-bisphosphate carboxyiase. Proc. Nati. Acad. Sci., U.S.A. 75: 6110-6114. 45. Lubben, T. H.; Theg, S. M.; Keegstra, K. 1988. Transport of proteins into chloroplasts. Photosyn. Res. 17: 173-194. 46. Mishkind, K. L.; Wessler, S. R.; Schmidt, G. W. 1985. Functional determinants in transit sequences: irnpot and partial 1 rnaturation by vascular plant chloroplasts of the ribulose-I, 5-bisphosphate carboxylase small subunit of chlamydornonas. J. Cell Biology 100: 266-300. 47. Schmidt, G. W.; Mishkind, M. L. 1986. The transport of proteins into chloroplasts. Ann. Rev. Biochem. 55: 879-912. 48. Kishore, G. M.; Shah. 1988. h i n o acid biosynthesis inhibitors as herbicides. Annu. Rev. Biochem. 57: 627-63. 49. Groten, J. P.; Schoen, E. D.; Van Balderen, P. J.; Kuiper, C. F.; Van Zorge, J. A.; Feron, V. J. 1997. Subacute toxicity of a mixture of nine chemicals in rats: detecting interactive effects with a fractionated hivo level factonal design. Fundam. Appl. Toxicol. 36: 15-29. 50. Jonker, D.; Wountersen, R. A.; Feron, V. J. 1996. Toxicity of mixture of nephrotoxicants with similar or dissimilar mode of action. Food Chem. Toxicol. 341075-1082. 51. Jonker, D.; Wountersen, R. A.; Van Bladeren, P. J.; Til, H. P.; Feron, V. J. 1990. 4-week oral toxicity of a combination of eight chemicals in rats: comparison with the toxicity of the individual compounds. Food Churn. Toxicol. 28: 623-631. 52. Jonker, et al.; 1993 Jonker, D.; Jones, M. A.; Van Bladeren, P. J.; Wourstersen, R. A.; Til, H. P. and Feron, V. J. Acute (24h) toxicity of a combination of four nephrotoxicants in rats compared with the toxicity of the individual compounds, Food and Chemical Toxicology 31(1993), pp. 42-52. 53. Hallas, L. E.; Hahn, E. M. e Komdorfer, C. 1988. Characterization of microbial traits associated with glyphosate biodegradation in industrial activated sludge. J. Industrial Microbiol. 3: 377-385. 54. Barry, G.; Kishore, G.; Padgette, S.; Taylor, M.; Kolacz, K.; Weldon, M.; Re, D.; Eichholtz, D.; Fincher, K.; Hallas, L. 1992. Inhibitors of amino acid biosynthesis: Strategies for imparting glyphosate tolerance to crop plants. In: Biosynthesis and Molecular Regulation of Amino Acids in Plants. Singh, B. K.; Flores, H. E. e Shannon, J. C. editors. American Society of Plant Physiologists. 139-145. 55. Rueppel, M. L.; Brightwell, B. B.; Schaefer, J. e Marvel, J. T. 1977. Metabolism and degradation of glyphosate in soil and water. J. Agric. Food. Chem. 25: 517-528. 56. Torstensson, L. 1985. Behavior of glyphosate in soils and its degradation. Pp. 137-150. In E. Grossbard and D. Atkinson (eds.). The Herbicide Glyphosate. Butterworth, London. 57. Giesy, J. P.; Dobson, S. e Solomon, K. R. 2000. Ecotoxicological risk assessment for Roundup® herbicide. Reviews of Environmental Contamination and Toxicology 167: 35-120 (Anexo 6 - 23). 58. Serdy, F. S. & Nida, D. L. 1995. Petition for determination for non- regulated status, cotton with the Roundup Ready gene, lines 1445 and 1698. Petition submitted to USDA/APHIS/BBEP on February 10, 1995. 59. Comissão Técnica Nacional de Biossegurança. CNTBio. 2009. Parecer Técnico 2041/2009. Publicado no DOU de 28/09/2009, Seção 1, página 21. 60. Huttner, S. L.; Arntzen, C.; Beachy, R.; Breuning, G.; Nester, E.; Qualset, C. and Vivalder, A. 1992. Revising oversight of genetically modified plants. Bio/ Technology 10, pp. 967-971. 61. Gruys, K. J. e Sikorki, J. A. 1999. Inhibitors of tryptophan, phenylalanine and tyrosine biosynthesis as herbicides. In Plant Amino Acids: Biochemistry and Biotechnology. (Ed. Singh, B.). pp. 357-384. Marcel Dekker Inc., New York, NY. 62. Alibhai, M. e Stallings, W. C. 2001. Closing down on glyphosate inhibition with a new structure for drug discovery. Proc. Nat’l Acad Sci USA 98: q 2944-6. 63. Franz, J. E.; Mao, M. K. e Sikorski, J. A. 1997.Glyphosate: A unique global herbicide. American Chemical Society (ACS), Washington, DC. ACS Monograph No. 189. 64. Harrison, L. A.; Bailey, M. R.; Naylor, M. W.; Ream, J. E.; Hammond, B. G.; Nida, D. L.; Burnette, B. L.; Nickson, T. E.; Mitsky, T. A.; Taylor, M. L.; Fuchs, R. L. e Padgette, S. R. 1996. The expressed protein in glyphosate-tolerant soybean, 5-enolpyruvyshikimate-3-phosphate synthase from Agrobacterium sp. strain CP4, is rapidly digested in vitro and is not toxic to acutely gavaged mice. J. Nutrition. 126: 728-740. 65. Fuchs, R. L.; Ream, J. E.; Hammond, B. G.; Naylor, N. W.; Leimgruber, R. M. e Berberich, S. A. 1993. Safety assessment of the neomycin phosphotransferase II (NPTII) protein. Bio/Technology 11: 1543-1547. 66. Astwood, J. D.; Fuchs, R. L. 1996. Allergenicity of foods derived from transgenic plants. In: Wuthrich, B.; Ortolani, C. (Eds.), Highlights in Food Allergy. Monographs in Allergy, vol. 32. Karger, Basel, pp. 105-120. 67. Astwood, James, D.; Leach, John, N. & Fuchs, Roy, L. 1996. Stability of food allergens to digestion in vitroNature Biotechnology 14, 1269-1273. 68. Bradford, K. J.; Deynze, A. V.; Gutterson, N.; Parrott, W.; Strauss, S. H. 2005. Regulating transgenic crops sensibly: lessons from plant breeding, biotechnology and genomics. Nature Biotechnology, 23(4): 439-444.
Upload:
Where detection method protocols and appropriate reference material (non-viable, or in certain circumstances, viable) suitable for low-level situation may be obtained:
molecular traditional methods
Relevant links to documents and information prepared by the competent authority responsible for the safety assessment: National Biosafety Commission
Upload:
Authorization expiration date: Not Applicable
E-mail:
gutemberg.sousa@mcti.gov.br
Organization/agency name (Full name):
National Biosafety Technical Commission
Contact person name:
Edivaldo Domingues Velini
Website:
Physical full address:
SPO Area 5 Qd 3 Bl B S 10.1 Brasilia DF
Phone number:
556134115516
Fax number:
556133177475
Country introduction:
The Brazilian National Biosafety Commission – CTNBio , is responsible to the technical decision on biological risk as a response to a request from the proponent. The technical decision is given on a definitive basis. Only the National Biosafety Council (CNBS) can revoke the decision (in case of commercial release), based on social-economical reasons and not on biosafety reasons. Once a decision is taken by CTNBio favorable to the commercial release of a new GMO (being it a plant or any other organism), CNBS has 30 days to issue a revoke. After these steps, the new product must be evaluated for conformity to the Brazilian standards by the registration and enforcement agencies (ANVISA – Ministry of Health, Ministry of Agriculture, Ministry of Environment and Ministry of Fisheries, according to the intended use of the product). If it conforms to the standards, it may be offered to the market. Every institution dealing with GMOs (including universities and public research institutes) has to have an Internal Biosafety Commission (CIBio), which is legally responsible of everything that may happen to be done or caused by the GMO
Useful links
Relevant documents
Stacked events:
At the discretion of, and upon consultation with, CTNBio, a new analysis and issuance of technical opinion may be released on GMOs containing more than one event, combined through classic genetic improvement and which have been previously approved for commercial release by CTNBio
Contact details of the competent authority(s) responsible for the safety assessment and the product applicant:
Dr. Edivaldo Domingues Velini (President of national Biosafety Commission)
Philippines
Name of product applicant: Monsanto Philippines
Summary of application:
Cotton 531 x Cotton 1445 includes two incorporated genes, cry1Ac and cp4epsps that encode proteins that convey protection from lepidopteran insect pests and tolerance to Roundup herbicide, respectively. The Cry1Ac protein is an insect control protein and acts through a toxic action in the gut of specific lepidopteran insects.

The cry1Ac gene was derived from the common soil bacterium, Bacillus thuringiensis subsp. kurstaki, and was introduced in the Bollgard cotton event 531. CP4 EPSPS protein belongs to the family of EPSP synthases, which are enzymes involved in the
penultimate step on the biochemical shikimate pathway producing aromatic amino acids in the chloroplasts of plants. The cp4 epsps gene inherited by Bollgard x Roundup Ready from the Roundup Ready cotton event 1445 as derived from Agrobacterium sp. Strain CP4, a common soil-borne bacterium.

A commercial variety with the introduced trait MON 1445 was developed by the traditional backcrossing of MON 1445 and the conventional variety. The resulting variety with MON 1445 was then crossed with another cotton line that contains MON 531. The resulting variety with the 1445 event was then crossed with another cotton MON 531. A minimum of five to six backcrosses was made to stabilize the introduced MON 531 with the commercial variety containing MON 1445. The
resulting seeds are stacked genes F1 hybrid (Bollgard x Roundup Ready cotton)
Upload:
Date of authorization: 08/01/2010
Scope of authorization: Food and feed
Links to the information on the same product in other databases maintained by relevant international organizations, as appropriate. (We recommend providing links to only those databases to which your country has officially contributed.):
Summary of the safety assessment:
Monsanto Philippines, has 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 combined trait product cotton: MON 531 x MON 1445 which has been genetically modified for insect resistance and herbicide tolerance. A safety assessment of combined trait product cotton: MON 531 x MON 1445 was conducted as per Administrative Order No. 8 Series of 2002 and Memorandum Circulars Nos. 6 and 8, Series of 2004. 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.
Upload:
Where detection method protocols and appropriate reference material (non-viable, or in certain circumstances, viable) suitable for low-level situation may be obtained:
Relevant links to documents and information prepared by the competent authority responsible for the safety assessment:
Upload:
Authorization expiration date:
E-mail:
bpibiotechsecretariat@yahoo.com
Organization/agency name (Full name):
Bureau of Plant Industry
Contact person name:
Thelma L. Soriano
Website:
Physical full address:
San Andres St., Malate, Manila
Phone number:
632 521 1080
Fax number:
632 521 1080
Country introduction:
The Philippines is the first ASEAN country to establish a modern regulatory system for modern biotechnology. The country's biosafety regulatory system follows strict scientific standards and has become a model for member-countries of the ASEAN seeking to become producers of agricultural biotechnology crops. Concerns on biosafety in the Philippines started as early as 1987 when scientists from the University of the Philippines Los Banos (UPLB) and International Rice Research Institute (IRRI), the Quarantine Officer of the Bureau of Plant Industry (BPI) and the Director for Crops of the Philippine Council for Agriculture, Forestry and Natural Resources Research and Development (PCARRD) recognized the potential for harm of the introduction of exotic species and genetic engineering. The joint committee formed the biosafety protocols and guidelines for genetic engineering and related research activities for UPLB and IRRI researchers. This proposal was eventually adapted into a Philippine Biosafety policy by virtue of Executive Order No 430, Series of 1990, issued by then President Corazon C. Aquino on October 15, 1990, which created the National Committee on Biosafety of the Philippines (NCBP). The NCBP formulates, reviews and amends national policy on biosafety and formulates guidelines on the conduct of activities on genetic engineering. The NCBP comprised of representative from the Department of Agriculture (DA); Department of Environment and Natural Resources (DENR); Health (DOH); and Department of Science and Technology (DOST), 4 scientists in biology, environmental science, social science and physical science and 2 respected members of the community. The Philippines’ Law, Executive Order No.514 (EO514), Series of 2006 entitled “Establishing the National Biosafety Framework (NBF), Prescribing Guidelines for its Implementation, Strengthening the National Committee on Biosafety of the Philippines, and for Other Purposes was also issued. This order sets the establishment of the departmental biosafety committees in the DA, DENR, DOH and DOST. The mandates jurisdiction and other powers of all departments and agencies in relation to biosafety and biotechnology is guided by the NBF in coordination with the NCBP and each other in exercising its power. The Department of Agriculture (DA) issued Administrative Order No 8, Series of 2002, (DA AO8, 2002), which is part of EO 514, for the implementation of guidelines for the importation and release into the environment of plants and plant products derived from the use of modern biotechnology. The DA authorizes the Bureau of Plant Industry (BPI) as the lead agency responsible for the regulation of agricultural crops developed through modern biotechnology. The BPI has adopted a protocol for risk assessment of GM crops for food and feed or for processing based on the Codex Alimentarius Commission’s Guideline for the Conduct of Food Safety assessment of Foods Derived from Recombinant-DNA plants and a protocol for environmental risk assessment in accordance with the Cartagena Protocol on Biosafety and with the recommendation of the Panel of Experts of the Organization for Economic Cooperation and Development (OECD). DA AO8, 2002 ensures that only genetically food crops that have been well studied and found safe by parallel independent assessments by a team of Filipino scientists and technical personnel from the concerned regulatory agencies of the Department are allowed into our food supply and into our environment. The DA AO 8, 2002 has a step by step introduction of GM plant into the environment. The research and development phase would require testing the genetically modified (GM) crop under controlled conditions subject to regulation by the government agencies. The first stage of evaluation for GM crops is testing under contained facilities such as laboratories, greenhouses and screenhouses. After satisfactory completion of testing under contained facilities, confined environmental release or field trial is done. Confined field trial (CFT) is the first controlled introduction of the GM crop into the environment. The approval for field trial shall be based on the satisfactory completion of safety testing under contained conditions. Unconfined environmental release or commercialization of the product would follow after the safe conduct of the CFT. Approval for propagation shall only be allowed after field trials and risk assessment show no significant risk to human and animal health and the environment.
Useful links
Relevant documents
Stacked events:
Gene stacking in plants can be conferred either through genetic engineering or conventional breeding A full risk assessment as to food and feed or for processing shall be conducted to plant products carrying stacked genes conferred through genetic engineering or conventional breeding, where the individual traits have no prior approval for direct use as food and feed or processing from the Bureau of Plant Industry (BPI) A desktop or documentary risk assessment on the possible or expected interactions between the genes shall be conducted for stacked gene products with multiple traits conferred through conventional breeding and individual events granted prior approval by the Bureau of Plant Industry.
Contact details of the competent authority(s) responsible for the safety assessment and the product applicant:
Bureau of Plant Industry 692 San Andres St, Malate, Manila 1004