Final Health and Environmental Risk Assessment of Genetically Modified Soybean MON 89788

Soybean MON 89788 expresses the cp4 epsps gene from the plant pathogenic bacterium Agrobacterium tumefaciens (Rhizobium radiobacter) sp. strain CP4. The encoded enzyme 5enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) protein confers tolerance to the active herbicidal substance glyphosate. Updated bioinformatics analyses of the inserted DNA and flanking sequences in soybean MON 89788 have not indicated a potential production of putative harmful proteins or polypeptides caused by the genetic modification. Genomic stability of the functional insert and consistent expression of the cp4 epsps gene, have been shown over several generations of soybean MON 89788. With the exception of the intended changes caused by the trans-genetically introduced trait, data from several field trials performed in USA and Argentina show that soybean MON 89788 is compositionally, morphologically and agronomically equivalent to its conventional counterpart and other commercial soybean varieties. A sub-chronic feeding study with rats, as well as a nutritional assessment trial with broilers has not revealed adverse effects of soybean MON 89788. These studies indicate that soybean MON 89788 is nutritionally equivalent to, and as safe as conventional soybean varieties. The CP4 EPSPS protein produced in soybean MON 89788 does not show sequence resemblance to known toxins or IgE-dependent allergens, nor has it been reported to cause IgE-mediated allergic reactions. Soybean is not cultivated in Norway, and there are no cross-compatible wild or weedy relatives of soybean in Europe. Based on current knowledge, the VKM GMO Panel concludes that with the intended usage, there are no discernible safety concerns associated with soybean MON 89788 regarding human or animal health or to the environment in Norway.

Standardization of PCR and sequencing-based methods for the detection of the presence of CP4 EPSPS gene in Zea mays

With the rise in planting of transgenic cultivars in Brazil as well as worldwide, the demand for legal regulations has increased. The transgenic event MON88017 is often found in maize cultivars marketed in Brazil. The event contains the CP4 EPSPS and cry3Bb1 genes, which encode tolerance to the herbicide glyphosate and resistance to caterpillars, respectively. Globally, methodologies to track transgenic events are mandatory. The objective of this study was to standardize a method based on qualitative PCR and sequencing for detection of the CP4 EPSPS gene in Zea mays. DNA was extracted from three commercial strains of transgenic maize, containing the MON88017 event, as well as conventional maize. Primers were designed for partial detection of the zein and CP4 EPSPS genes. PCR reactions were performed for detection of partial regions of CP4 EPSPS and Zein (as endogenous marker of Z. mays) genes. The three transgenic maize inbred lines tested positive for zein and CP4 EPSPS, and the two conventional strains tested negative for CP4 EPSPS and positive for the Zein gene. To confirm the presence of the genic regions, PCR products were sequenced and showed 100% identity with sequences of Zein and CP4 EPSPS genes deposited in GenBank. Thus, the results of this study suggest the applicability of an ‘in-house’ method for the qualitative detection of CP4 EPSPS in genetically modified maize cultivars.

A necessity of introduction of the system of registration of GMO is for Ukraine

The article presents the main methodological approaches and criteria for the necessity of development and introduction of the State Registration of GMOs sources in Ukraine. The list of analytical researches that are to be carried out at the registration of the GMO source, expert evaluation of GMO identification methods, reproduction of detection methods, identification and quantification of GMOs are given. Submitted and analyzed legislative documents regulating the procedure for registration and control of GMO circulation in the countries of the European Union and Ukraine. DNDKI of veterinary preparations and fodder supplements on the tasks of the State Committee of Veterinary Medicine of Ukraine during 2010-2018 a number of normative documents were developed regarding procedures related to the use and circulation of feed, feed additives, premixes and veterinary preparations that are composed, containing or made using genetically modified sources in Ukraine: the procedure for state registration, the procedure for carrying out the state veterinary and sanitary examination of GMOs for sources of forage feed additives and veterinary preparations ativ plan state monitoring GMO sources of feed, feed additives, premixes for livestock and poultry. The procedure determines the procedure for state registration of GMOs for sources of feed, feed additives and veterinary preparations containing genetically modified organisms or obtained from their use. It is envisaged that the state register of GMO sources is conducted in the form approved by the State Service of Ukraine for Food Safety and Consumer Protection. On the basis of the analysis, a draft list of feeds, feed materials, protein-vitamin concentrates and premixes that have to be monitored for GMO content has been developed. This project list includes soy, corn, rape and products of their processing, fodder, feed mixes, feed concentrates, etc. In the SSRCI of veterinary drugs and supplements, in 2018, the “Guidelines for identifying the GM soya line (Soybean MON40-3-2) GTS 40-3-2” were developed and approved. “Methodological recommendations are intended to identify the GM soya line (Soybean MON40-3 -2) GTS 40-3-2 (Genetically modified soybean that contains – genetically modified soybean containing (cp4 epsps gene inserted to confer tolerance to herbicide glyphosate) cp4 epsps gene inserted to provide resistance to glyphosate herbicide (glyphosate) in plant material, feed and food products by polymerase chain reaction in real time (PCR-RF )”. The qualitative determination of GMOs is based on the discovery of genetically modified (GM) targeted regulatory sequences of the 35S Cauliflower Mosaic Virus (CaMV) promoter or FMV promoter and/or the NOS (T-NOS) T1 terminator of Agrobacterium tumefaciens plasmids that are introduced into the genetic constructs of GM based on the use of the multiplex polymerase chain reaction method to obtain real-time research results – PCR RF.

Development of Gene-Specific Real-Time PCR Screening Method for Detection of cp4-epsps Gene in GM Crops

Among the genetically modified (GM) crops that are being approved for commercialization, herbicide resistant crops, especially those harboring cp4-epsps, have a considerable contribution. Gene-specific methods can be used to screen the presence of GMOs. To establish an effective qualitative and quantitative screening method, a set of primers were designed considering the cp4-epsps sequence. The specificity, the limit of detection, the efficiency, and the ability to quantify the GMO content were tested in GM cotton, soybean, and canola events. The results demonstrated that the primers can specifically detect cp4-epsps GM crops. The limit of detection was found to be 0.4 ng /μl DNA per PCR reaction with the ability to detect 1-16 copies of the haploid genome of each GM event. The efficiency of this screening method (which was 94-110 % with an R2 higher than 0.96) indicated that these new primers can be applied to the screening of GM samples that contain the cp4-epsps gene. Also, the gene-specific real-time PCR screening method could be successfully developed for qualification of different types of GM cotton, soybean and canola events with the construction of a serial dilution ranging from 10 % to 1 %.