• Inheritance studies conducted indicated that Mendelian segregation exists. • New expression proteins are expressed in low levels. • It is compositionally equivalent to its non-transgenic counterpart. • No evidence of similarity or homology was found with known toxic proteins. • There is no evidence of expression of known allergenic substances for the proteins expressed in the event. It is concluded that the event is substantially equivalent to its conventional counterpart, therefore, it is as safe and no less nutritious than conventional commercial varieties.

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

Please refer to the Risk Assessment Report

Authorization by COFEPRIS: 176

Maize (Zea mays) genetically modified MZHG0JG with resistance to glufosinate-ammonium and glyphosate herbicides.

No documentary evidence indicating toxicological and / or allergenic effects was observed;as well as substantial nutritional changes in the Maize MZHG0JG, so it is concluded that the maize (Zea mays) genetically modified with OECD Identifier SYN-∅∅∅JG-2 is, based on the existing knowledge to date, as innocuous as its conventional counterpart.

Simplified approval procedure: Through MAG’s Resolutions 1030 and 1071 there was stated a differentiated treatment for the commercial release of novel GE crops and for GE crops that have been approved in third countries, whose scientific, technical and safety characteristics are well-founded. Paraguayan Ministry of Agriculture’s Resolutions authorize taking into consideration the decision documents from third countries with regard to both human and animal food safety in the cases where these evaluations have been based on Codex Alimentarius, such as the Guidelines for the Conduct of Food Safety Assessment of Foods Derived from Recombinant-DNA Plants and carried out in countries with time-tested regulatory systems and transparent procedures. Concerning environmental safety, assessments are accepted for GE crops that besides having been authorized for commercial planting in countries with sound regulatory systems, include in the decision documents considerations as follows: that the GE crop under review has been studied under different environmental conditions, behaving in the same way as the conventional non-GE counterpart; that it will be managed in an agronomic manner similar to any GE or conventional hybrid/variety of the species; another aspect is that Paraguay is not center of origin of that crop, and finally two relevant characteristics are that there are no related weeds in Paraguay with which the GE crop could cross-breed and that the main target pests and the main non-target arthropod species present in Paraguay have been taken into account in the GE risk assessment carried out in those countries. The Commercial Release Opinion of the National Commission for Agricultural and Forestry Biosafety (CONBIO), in its substantial part states: "...Recommends technically: 1) The commercial release of the event MHG0JG 2) In case of detection of an unexpected effect, the company is obliged to inform CONBIO".

MZHG0JG corn has been adequately assessed to demonstrate its safety as food and feed. Compositional and nutritional analyses show that MZHG0JG corn is nutritionally and compositionally similar to, and as safe as conventional corn. Studies conducted show that the proteins produced in the MZHG0JG corn (mEPSPS and PAT) do not have homology to any known mammalian toxin or allergen. There is no evidence that these proteins will interact to form new toxin or allergen. In addition, Southern blot analyses demonstrated that the MZHG0JG is stably inherited from one generation to the next.

 

Therefore, based on compositional/nutritional analyses, allergenicity and toxicity studies specifically conducted to assess its safety and from the data available from the scientific literature, MZHG0JG corn is substantially equivalent to and is as safe as the conventional corn, except for its built-in tolerance to glyphosate and glufosinate herbicides. Furthermore, the well-characterized modes of action, physiochemical properties, and a history of safe use demonstrate that the mEPSPS and PAT proteins present in MZHG0JG corn present no risk of harm to humans or livestock that consume corn products.

 

MZHG0JG corn will only be imported for direct use as food, feed, and for processing and will not be propagated in the Philippines. In the event that there is an accidental release of grains in the environment during importation, chances of survival of MZHG0JG corn grains is unlikely since it will generally occur in industrial areas that are highly disturbed habitats unsuitable for significant plant establishment. The spilled grain has to reach the soil and be exposed to specific favorable conditions, such as light, moisture, and temperature so that the germination and plant establishment to occur.

 

To date, MZHG0JG corn has passed the thorough evaluation of various regulatory institutions and has received approvals for propagation or release into the environment in 2 countries and approvals for import in 4 countries.

 

MZHG0JG corn will be imported as grains for direct use as food, feed, and for processing in the Philippines.  Since MZHG0JG corn will be a traded commodity, the society will be the main beneficiary in case importation will be required to supply the industry with grains of corn and corn products to feed millers, for use in poultry, livestock, aquaculture, and other industries.

 

STRP, BAI, BPI-PPSSD Assessment and Recommendation Based on the documents submitted by the applicant: A. Host Organism The assessors concurred that corn is a good source of carbohydrates, fats, calcium and phosphorus. It has long been domesticated and grown worldwide and is the second most important cereal crop in the Philippines. Corn has phytic acid which reduces the availability of phosphorus in monogastric animals. Raffinose and trypsin inhibitors are also present in corn. A potential toxicant in roots and leaves, DIMBOA (2-4-hydroxy-7 methoxy-2H-1,4- benzoxacin-3 (4H)-one, has been reported and its level among corn varieties has been varied and its amount decreases as the corn grows. However, amino acid sequence alignment did not match with any protein toxicant in the NCBI and Syngenta database. B. Transgenic Plant The corn event has been approved as food and feed in US and Canada, and as food in New Zealand. The assessors agreed with the report that the transgenic plant is not a source of allergens. This was further confirmed by the results of gastric fluid digestibility studies, SDS-PAGE and Western blot analyses. Consumption of corn as food may not drastically change due to the availability of transgenic corn varieties like MZHG0JG corn. Assessment from these countries proved that the transgenic corn is not materially different in composition, safety, and other relevant parameters from corn-derived food and feed found in the market. C. Donor Organism The two protein-encoding genes namely, mepsps-02 (modified 5-enol pyruvylshikimate-3-phosphate synthase gene) and pat-09 (phosphinothricin acetyltransferase gene) have been properly described with respect to source and potential pathogenic and allergenic properties. The gene mepsps-02 from Zea mays encodes the enzyme mEPSPS, while pat-09 gene from Streptomyces viridochromogenes, a common soil bacterium, encodes PAT protein. All genetic elements that make up the gene cassette have been elaborated by the proponent. The MZHG0JG insert and its flanking sequences are intact with no changes in base pairs. D. Transformation System The transformation method used was Agrobacterium tumefaciens mediated transformation, producing MZHG0JG. Genetic elements within the border regions of the T-plasmid were integrated into the plant genome, while those outside the borders were not transferred. The target of modification was nuclear DNA and the complete details of the experimental protocol were provided, including a complete description of the plasmid vector. Neither carrier DNA nor helper plasmids were detected. E. Inserted DNA Genetic Stability The integrity of the genetic elements within the insert was also evaluated in succeeding generations of transgenic corn plants. Stability of the genetic elements in the insertion sites were evaluated through Southern blot analysis and application of the probe for the plasmid backbone utilizing two corn generations. The absence of extraneous T-DNA fragments in the genome and in the backbone sequences of the transformation plasmid vector of the transgenic MZHG0JG was described supporting the integrity of the genetic elements within the insert. Open reading frames were not identified for comparison with allergen and toxin data base. These data conform with the conclusion that no amino acid translations of the start-to-stop junction open reading frames share relevant amino acid sequence similarity to known or putative protein allergens and toxins. The mEPSPS integrated in the MZHGOJG corn has been expressed in other transgenic corn varieties like GA21corn. The similarity of the mepsps-02 genes that encode for the mEPSPS protein in MZHGOJG corn and GA21 corn were validated by nucleotide sequencing. F. Genetic Stability Stability of the insert has been validated by Southern blot analysis which demonstrated identical hybridization bands for the MZHGOJG insert in lanes where the DNA extracts of transgenic plants grown for five generations. The presence of the mepsps-02 and the pat-09 genes in three generations of the transgenic corn was confirmed by the data generated from Real-time PCR. Chi-square analysis was used to validate the heritability of the MZHG0JG insert in a predictable manner in line with Mendelian principles and the results generated validated the previous hypothesis. G. Expressed Material Concentrations of proteins mEPSPS and PAT in MZHG0JG leaves, roots, whole plants, pollen, and kernels were measured by Enzyme Linked Immunosorbent Assay (ELISA). The two proteins are both enzymes and therefore participate in specific metabolic pathways. The native EPSPS in corn catalyzes the synthesis of 5-enoylpyruvylshikimate-3-phosphate (EPSP) from shikimate-3-phosphate (S3P) and phosphoenolypyruvate (PEP). This reaction is one of the steps in the synthesis of aromatic amino acids. The herbicide glyphosate inhibits the action of the native enzyme making the native corn incapable of producing aromatic amino acids which leads to its death. On the other hand, the modified EPSP (mEPSP) in MZHG0JG differs with the native enzyme in that it contains two amino acid substitutions that make the modified enzyme lose its affinity with glyphosate thereby making the transgenic corn tolerant to the herbicide. Simulated gastric fluid at pH 1.2 and the T50 is practically none because all got inactivated at one minute. Determination was done using SDS-PAGE. Pepsin was used in simulated gastric fluid studies and an intact mEPSPS was not detected after digestion for one minute, while faint diffused bands with low molecular weight were visible in SDS-PAGE analysis of the digested protein. The bands with low molecular weight did not cross-react with anti-mEPSPS antibody when assessed with Western blot. These reported observations demonstrate that mEPSPS in transgenic maize or in recombinant E. coli readily degrade under simulated mammalian gastric conditions. The quick degradation was proven by the absence of intact mEPSPS upon sampling even at one min and that no immuno-reactive protein of mEPSPS is detected. These imply that mEPSPS is readily digested as a conventional dietary protein in normal mammalian gastric environment. H. Toxicological Assessment The assessors confirmed that pepsin was used to simulate gastric fluid digestibility studies and an intact mEPSPS was not detected following digestion for one minute. SDS PAGE revealed the presence of faint diffuse bands of lower molecular weight over time and Western blot analysis showed that this band did not cross react with the anti-mEPSPS antibody. It was reported that subjecting the microbially-produced and the MZHG0JG mEPSPS proteins at lower temperature (25 to 37°C) for 30 minutes reportedly caused minimal reduction in mEPSPS immunoreactivity (96 and 92%, respectively). Incubation of the sample at higher temperature (65 to 95°C) for 30 min lead to non-stability of mEPSPS and complete loss of immunoreactivity when assessed by ELISA. Data demonstrate that mEPSPS is stable for 30 min when heated up to 37°C. At temperatures above 65°C, which are often applied in the processing and cooking of corn, minute and negligible amounts of intact and functional mEPSPS protein in foods and feed may be expected. The assessors also reported that systematic comparison of the mEPSPS amino acid sequence with those of the sequences of known or putative protein toxins in the NCBI Entrez Protein database (NCBI version 2016) and in the Syngenta Toxin database (version 2016) with the use of BLAST confirmed that mEPSPS is neither a toxic protein nor it shares significant sequence similarity with other known or putative toxins. Administration of mEPSPS in mice at a dose of 2000 mg/kg body weight through oral gavage was not toxic. Treatment-related effects such as alteration in food consumption, hematology parameters, blood clinical chemistry parameters, post-mortem alteration of weights of organs were not significantly different between mice in the treated (mEPSPS-gavaged groups) and control groups. The assessors also confirmed that the susceptibility of PAT to proteolytic degradation was evaluated in simulated gastric fluid (SGF) containing pepsin and the results showed that PAT is readily digested in SGF in less than one minute as assessed by SDS-PAGE and Western blot. There are no fragments left after digestion in SGF as assessed by SDS-PAGE and Western blot analyses. It can therefore be concluded that PAT is readily digested by the mammalian gastric enzyme, pepsin. It was also reported that immunoreactivity of PAT is no longer detectable after incubation for 30 minutes at 95˚C. The results of BLASTP showed no significant homology of PAT to any known toxin. NCBI Database Alignments identified 1000 sequences with potentially significant similarity to the PAT amino acid sequence. Of these, 651 proteins were identified as PAT proteins; 275 were identified as acyl transferase proteins; 73 proteins were identified as hypothetical/ unknown proteins. One protein was identified as a DNA alkylation repair protein from Streptococcus suis. The toxicity of the PAT protein administered two times a day by oral gavage in mice at a total dose level of 2000 mg/kg was evaluated. A 14-day observation was well-tolerated with no evidence of toxicity. Animals that received the gavage survived until the scheduled time for necropsy. No test substance-related clinical effects on body weight, food consumption, gross and macroscopic findings were reported. The assessors affirmed that the test protein was produced in Escherichia coli expression system and is therefore microbially-produced. Equivalency was demonstrated through Western blot, glycosylation, and peptide mass coverage analyses. The microbially-produced protein was compared to plant-produced PAT. Western blot analysis showed that the two proteins exhibit a single immunoreactive band consistent with the predicted molecular weight of 20.5 kDa. Glycosylation analysis showed no evidence of post-translational glycosylation for both proteins Peptide mass coverage analysis mapping confirmed the identity of the proteins from both sources as PAT. Furthermore, the assessors have reported that mEPSPS and PAT are expressed independently of each other to retain their functional properties. The mEPSPS and PAT are integrated within the corn nuclear genome. Expression of the proteins were validated and confirmed by a series of tests that applied Southern blot, nucleotide sequencing of the nuclear genome and examination of inheritance pattern of the transgene over three generations of the transgenic MZHG0JG corn. The assessors confirmed that the expressed proteins do not interact with each other. They act as independent system conferring tolerance to herbicides of glufosinate or glyphosate composition I. Allergenicity Assessment The assessors confirmed that the Digestibility studies that used pepsin (2600 units/ml) was performed at a ratio of 10 pepsin activity units/µg EPSPS. No intact mEPSPS (ca. 47.4 kDa) was detected following one min digestion in SGF. Faint diffused bands that registered low molecular weight were visible in SDS-PAGE over time. The bands did not cross-react with anti-mEPSPS-antibody in Western blot. These data demonstrate the fast degradation of the mEPSPS expressed in a consumed transgenic corn by mammalian gastric conditions. There was loss of immune reactions for mEPSPS at 25⁰C and 37⁰C incubation for 30 mins with 92-96% loss of immune reactivity and complete loss at 95 ⁰C incubation at 30 minutes. This was determined through ELISA. It was reported that the mEPSPS protein showed no sequence similarity with any known allergen. The mEPSPS protein is not homologous with known allergens as demonstrated by the absence of similar sequences (indicated by greater than 35% shared identity over 80 or more amino acids) when compared with the amino acid sequences of any allergen in the FARRP allergen online database. The absence of similar alignments of eight or more contiguous amino acids between the mEPSPS and sequences of allergens in the FARRP allergen database. On the other hand, the digestibility of PAT to pepsin was done, which showed rapid digestion in less than oneminute, as assessed by SDSPAGE and Western blot. As the digestion appeared complete, there were no fragments detected by SDS-PAGE and Western blot. A diminishing immuno-reactivity is reported in PAT at 55.2%, 11.2% and 5.4% relative to its incubation at 25, 37 and 65°C, respectively. Incubation at 95°C for 30 min contributed to the low detection of immune-reactivity of PAT when evaluated by ELISA. Data indicate that a complete loss of detectable immune reaction of PAT is contributed by heating at high temperature which parallels possible inactivation of PAT through heating and cooking of corn. The FASTA search did not show homology of greater than 35% of shared identity for the 80 aa bases. No biologically relevant match was found for allergens. PAT proteins are microbiologically derived for the transgenic event. These have not been shown to be glycosylated. Comparison of the plant and microbial derived PAT shows the same mol. wt within the range of 10-70 kDa range. J. Nutritional Data For the proximate analysis, the assessors confirmed that the comparison of the transformed corn forage and conventional counterpart showed equivalency of moisture, protein, fat, ash, acid detergent fiber (ADF) and neutral detergent fiber (NDF). However the transformed plant has a higher carbohydrate relative to the conventional corn. Furthermore, six non-transgenic commercial corn varieties were included in the comparison. All the control, MZHG0JG, MZHG0JG + trait specific herbicide (TSH), and the six commercial varieties were grown in ten corn-growing locations in the US in June 2013. The assessors reported that all the data on forage proximates derived from the transgenic lines are within the observed range. The assessors confirmed that based on the results of the analysis provided by the developer, no statistical differences in the proximates of forage corn were observed that can be considered as biologically relevant. On the other hand, the assessors confirmed that the applicant provided sufficient information regarding the proximate analysis of MZHG0JG grains in comparison with the non-transgenic corn. Levels of all proximates in MZHG0JG showed no significant difference with the non-transgenic corn except for NDF which was significantly lower than the control. In terms of MZHG0JG corn treated with TSH, no significant differences were observed in the levels of protein, fat, ash, carbohydrates, ADF, TDF, NDF and starch in comparison with the non-transgenic corn. Moreover, it was reported that in MZHG0JG corn, and MZHG0JG corn + TSH, the mean levels of all proximates were within the ranges for the reference varieties. It was confirmed that while there was statistical significance on NDF, the difference was minimal in favor of better forage. Overall, the values fall within the range of commercial and ILSI values which provide proof of equivalency in the proximate analysis results. In key nutrients analysis of forage, a corresponding non-transgenic, near-isogenic control corn was used as a comparator. Statistically, there are no significant differences between MZHG0JG, MZHG0JG + TSH and the control in terms of the amounts of calcium and phosphorus in forage. Furthermore, in reference to ILSI Database, mean levels of calcium and phosphorus in MZHG0JG corn and MZHG0JG corn + TSH were also within the ranges. Again, it was confirmed that based on the results of the analysis, no statistical differences in mineral composition of forage MZHG0JG were observed that can be considered as biologically relevant. In key nutrients analysis of grain, a corresponding non-transgenic, near-isogenic control corn was used as a comparator. Statistically, there are no significant differences between MZHG0JG, MZHG0JG + TSH and the control in terms of the amounts of calcium, magnesium, manganese, phosphorus, potassium, selenium, sodium, and zinc. The amounts of copper and iron were significantly lower in the two transgenic plants compared to the control. Furthermore, between MZHG0JG corn and the non-transgenic control corn, levels of vitamins B1, B2, B3, and B9 have no significant differences; level of vitamin A was significantly higher than in the control; and levels of vitamins B6 and E were significantly lower than in the control. Between MZHG0JG corn + TSH and control, no statistically significant differences were observed in the levels of B2, B3, B6 and B9;; on the other hand, levels in B1 and E was lower than in the control. The assessors reported that based on the results of the analysis provided by the developer, no statistical differences between the mineral and vitamin content of MZHG0JG grains were observed.. In addition, the assessors reported that no significant differences were observed in the levels of 15 amino acids between MZHG0JG corn and the non-transgenic control corn. The levels of aspartic acid, arginine, and tryptophan were significantly lower in MZHG0JG corn than in the control corn. Between MZHG0JG corn and control corn, no statistically significant differences were observed in levels of 14 amino acids. The levels of aspartic acid, lysine, arginine, and tryptophan were lower in MZHG0JG corn + TSH than in the control corn. Furthermore, as shown in the results of the analysis, there is no significant difference between the proportions of 16:0 palmitic, 16:1 palmitoleic, 18:0 stearic, 18:1 oleic, 18:2 linoleic, 20:0 arachidic, 20:1 eicosenoic, and 22:0 behenic fatty acids in MZHG0JG compared to the non-transgenic comparator. The proportions of 17:0 heptadecanoic acid and 18:3 linoleic fatty acid were significantly higher in MZHG0JG than in the non-transgenic comparator. In terms of the treatment with TSH, MZHG0JG showed higher proportions of 17:0 heptadecanoic and 18:3 linolenic fatty acids and lower proportions of 16:0 palmitic fatty acids compared to non-transgenic corn. The assessors concluded that based on the results of the analysis provided by Syngenta regarding the amino and fatty acid levels of MZHG0JG and non-transgenic comparator, no statistical differences were observed. For the analysis of antinutrients, there is no significant difference between the levels of ferulic acid, inositol, phytic acid, trypsin inhibitor and raffinose in MZHG0JG compared to the non-transgenic comparator. The level of ρ-coumaric acid observed was significantly higher in MZHG0JG than the non-transgenic corn. The same was observed in MZHG0JG + TSH including inositol which also exhibited significantly higher level than the non-transgenic comparator. In the MZHG0JG corn, and MZHG0JG corn + TSH, the mean levels of anti-nutrients were within the ranges for the reference varieties. In reference to ILSI Database, mean levels of anti-nutrients in MZHG0JG corn and MZHG0JG corn + TSH were within the ranges. Based on the results of the analysis, no statistical differences between the levels of anti-nutrients of MZHG0JG and the non-transgenic corn were observed. K. Recommendation After a thorough scientific review and evaluation of the documents provided by the Bureau of Plant Industry (BPI) to the STRP, BAI, and BPI-PPSSD, the assessors found scientific evidence that the regulated article applied for human food and animal feed is as safe as its conventional counterpart and shall not pose any significant risk to human and animal health.

Please see the link below(in Korean).

There were submitted (1) data enabling to identify the matter of research (species, variety, and the transformation event); (2) data on the initial parental organism and the donor organism for introduced genetic sequences;  (3) data on the genetic modification method, genetic construction, and the level of gene expression; (4) data on identification of GM maize line MZHG0JG (identification methods, protocol of analysis, description of primers, reference materials); (5) data on registration of the GM line  in other countries and the results of safety assessment which conducted for registration purposes of GMO in other countries.

 

Peer review of the data submitted by the applicant and the results of complex medical and biological studies of transgenic maize line MZHG0JG tolerant to glyphosate and glufosinate herbicide, attest to the absence of any toxic, reprotoxic, genotoxic, or allergenic effects of this maize line. By biochemical composition, transgenic maize line MZHG0JG was identical to conventional maize. GM maize line MZHG0JG has been registered for food use, listed in the State Register, and licensed for use in the territory of The Eurasian Economic Union, import into the territory of The Eurasian Economic Union, and placing on the market without restrictions.

Corn line MZHG0JG (SYN-000JG-2) has been modified to tolerate the herbicides glyphosate and glufosinate ammonium (glufosinate). Tolerance to glyphosate is achieved through expression of the enzyme 5-enolpyruvyl-3- shikimatephosphate synthase (mEPSPS) encoded by a modified epsps gene (mepsps-02, hereafter referred to just as mepsps) derived from corn (Zea mays). Tolerance to glufosinate is achieved through expression of the enzyme phosphinothricin acetyltransferase (PAT) encoded by the pat-09 gene derived from Streptomyces viridochromogenes. MZHG0JG was generated through Agrobacterium-mediated transformation and contains two expression cassettes. Molecular analyses indicate there is a single insertion site comprising a single, complete copy of each of the mepsps, and pat-09 genes together with their regulatory elements. The introduced genetic elements are stably inherited from one generation to the next. Corn line MZHG0JG expresses two new proteins, mEPSPS and PAT, which lack toxic or allergenic potentials. Detailed compositional analyses were done of proximates, fibre, minerals, amino acids, fatty acids, vitamins, secondary metabolites and anti-nutrients in grain taken from MZHG0JG given two treatments (herbicide-sprayed and unsprayed). Only 16 of the 58 analytes that were reported deviated from the control in a statistically significant manner; for five of these the difference occurred only in one of the MZHG0JG treatments. However, the mean levels of all of these analytes fell within both the reference range and the historical range from the literature. With the exception of vitamin A, the differences between these statistically significant analytes means of MZHG0JG and the control means were smaller than the variation within the control. Food derived from Corn line MZHG0JG is as safe as food derived from its non-genetically modified counterpart.

Commodity:Corn / Maize (Zea mays L.)

 

Maize event MZHG0JG has been genetically modified to expresses mEPSPS protein which confers tolerance to glyphosate herbicide and enzyme phosphinothricin N-acetyl transferase (PAT) which confers tolerance to glufosinate-ammonium herbicide.
PAT protein used 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 maize event MZHG0JG is substantially equivalent to its conventional counterpart in terms of morphology, nutrition, toxicity and allergenicity.