GM Plants Research Articles

Disharmonization in the Regulation of Transgenic Plants in Europe

Disharmonization in the Regulation of Transgenic Plants in Europe

Environmental Risk Assessment of Insect Resistant Genetically Modified Maize MON810 for Cultivation, Seed Production, Import, Processing and Feed Uses under Directive 2001/18/EC (Notification C/F/95/12/02)

Environmental Risk Assessment of Insect Resistant Genetically Modified Maize MON810 for Cultivation, Seed Production, Import, Processing and Feed Uses under Directive 2001/18/EC (Notification C/F/95/12/02)

Preliminary Environmental Risk Assessment of Insect Resistant Genetically Modified Maize MON 89034 for Cultivation (EFSA/GMO/BE/2011/90)

Preliminary Environmental Risk Assessment of Insect Resistant Genetically Modified Maize MON 89034 for Cultivation (EFSA/GMO/BE/2011/90)

Preliminary Environmental Risk Assessment of Genetically Modified Oilseed Rape MON 88302 for Food and Feed Uses, Import and Processing under Regulation (EC) No 1829/2003 (Application EFSA/GMO/BE/2011/101)

Preliminary Environmental Risk Assessment of Genetically Modified Oilseed Rape MON 88302 for Food and Feed Uses, Import and Processing under Regulation (EC) No 1829/2003 (Application EFSA/GMO/BE/2011/101)

Environmental Risk Assessment of Glufosinate-Tolerant Genetically Modified Oilseed Rape T45 for Food and Feed Uses, Import and Processing under Regulation (EC) No 1829/2003 (Application EFSA/GMO/UK/2005/25)

Environmental Risk Assessment of Glufosinate-Tolerant Genetically Modified Oilseed Rape T45 for Food and Feed Uses, Import and Processing under Regulation (EC) No 1829/2003 (Application EFSA/GMO/UK/2005/25)

Environmental Risk Assessment of Glufosinate-Tolerant Genetically Modified Oilseed Rape MS8, RF3 and MS8 x RF3 for Import, Processing and Feed Uses under Directive 2001/18/EC (Notification C/BE/96/01)

Environmental Risk Assessment of Glufosinate-Tolerant Genetically Modified Oilseed Rape MS8, RF3 and MS8 x RF3 for Import, Processing and Feed Uses under Directive 2001/18/EC (Notification C/BE/96/01)

Detection of mPing mobilization in transgenic rice plants.

Various kinds of transposable elements (TEs) constitute high proportions of eukaryotic genomes. Although most of these TEs are not actively mobile, genome stress can induce mobilization of dormant TEs. Transgenic plants undergo tissue culture and subsequent whole-plant regeneration, which can cause genomic stress and in turn induce mobilization of inactive TEs. To investigate the activation of transposable elements on the genome wide of the GM plant. Transposon activities were analyzed in three transgenic rice plants carrying the insect resistance gene Cry1Ac and an herbicide resistance gene by the transposon display technique. These three transgenic plants were derived from a leading Korean rice variety, Illmi. We detected seven mobile activities in the mPing element, which is a MITE family transposon. The identity of the novel fragments in the gel display was confirmed by checking TAA target site duplication via sequence analysis. The genomic integration sites were all on different chromosomes, and the integrations were specific to either one or two T1 transgenic lines, except for one common integration on chromosome 4. One integration was in the 5'-UTR of the Glycerol-3-phosphate acyltransferase 8 gene, two integrations were in introns of expressed genes, and the other four integrations were in intergenic regions. Thus, novel mobilization of dormant TEs occurs in transgenic plants, which must be considered in the generation of genetically modified crops (GM crops).

The Companion effect between Salicylic acid and Glutamine on the growth and yield of water-stressed cucumber (Cucumis sativus L. )

Trial was conducted in greenhouse of the Department of biology / College of Science/University of Diyala during autumn season 2016. The study included two levels of water stress ( S1/Irrigation for 14 minutes and S2 / Irrigation for 7 minutes/7 ml of water per minute ), three foliar application levels of Salicylic acid SA (0, 75, 150) mg. L−1 and three foliar application levels of Glutamine acid G (0, 1000, 2000) mg. L−1. Some growth indicators viz., (Plant height, Stem diameter, Relative growth rate and Plant Yield were registered. The experiment was designed as a factorial experiment using an RCBD design with three replicates. Water stress decreased the means of all studied indicators, Plant height record 232.44 cm, Stem diameter 8.28mm, Relative growth rate 0.019 g.day−1 and yield 1358 gm plant−1 at the first stress level (Irrigation for 14 minutes / S1) while the means of the indicators decreased to183.82 cm, 7.41 mm, 0.016 g day−1 and 972 g plant−1, respectively at the second stress level (Irrigation for 7 minutes / S2). Salicylic acid and Glutamine acid increased all the studied indicators. the third level of salicylic and Glutamine spraying (150 mg. L−1, 2000 mg. L−1 ) has exceeded the second level (75 mg. L−1, 1000 mg. L−1 ). The role of Salicylic acid and Glutamine acid was positive to decrease the passive effect of water stress, Plant height record 236.17 cm, Stem diameter 8.57mm, Relative growth rate 0.021 g.day−1 and yield 1893 gm plant−1(the percentage of increase was 37.57%, 40.95%, 50.00%, 179.62%, respectively), also the triple interaction between the second stress level(S2) and the spraying at high levels the Salicylic acid and Glutamine SA3 G3 was positive to decrease the effect of water deficit stress, 262.00 cm, 9.22 mm, 0.024g.day−1 and yield 2317gm plant−1 (percentages of increase were 75.05%,58.97%,100.00%and286.17%, respectively).

Delivery of CRISPR/Cas Components into Higher Plant Cells for Genome Editing

CRISPR/Cas genome editing of plants is realized in three basic variants, including knockout mutations as indels, insertion of alien DNA fragments, and base editing via deamination of nitrogenous bases. The most important stages of the CRISPR/Cas-based genome editing are the choice of a target site, design of guide RNAs, creation of genetically engineered constructions, and delivery of CRISPR/Cas components into a plant cell. Rapid developments in the field of plant genome editing with the use of CRISPR/Cas systems requires more detailed consideration of the last stage, so this review is dedicated to the description of the main ways to deliver CRISPR/Cas components into cells of higher plants. In the first studies on the genome editing of different plant species, these components were delivered to the target site mainly by Agrobacterium tumefaciens. This approach supposes integration of T-DNA into a genome and a stable expression of CRISPR/Cas components or their transient expression in the case of agroinfiltration. Another widespread approach included the use of plant viruses as delivery platforms; in this case, viruses were used mainly for production of an increased amount of guide RNAs that significantly improved the efficiency of genome editing. Another approach provides for the use of another bacterium, A. rhizogenes, as a platform for delivery of CRISPR/Cas components. This bacterium induces hairy root formation that may be an indirect confirmation of successful genome editing and assist in the selection of genetically modified forms. Other common ways to obtain genetically edited plants are the biolistic delivery of genetically engineered constructions into explants and various protoplast transformation technologies. The review also considers some issues transgenic and GM status of CRISPR/Cas-edited plants to transgenic and GM plants. There are a number of cases in which new organisms created by a CRISPR/Cas genome editing without any introduction of alien DNA were not considered as transgenic ones; it is quite possible that such plants will not fall under Russian legislation prohibiting GMO cultivation.

Effect of transgenic soybean on functional groups of microorganisms in the rhizosphere in soil microcosm

Neglected Glyphosate-resistant (GR) crops are common in agro-ecosystems mainly due to its benefits of weed management. However, the effect of GR crops on soil ecosystem and on non-target soil organisms need to be monitored. The effect of two transgenic soybeans GR on soil microorganisms, soil enzymes, microbial biomass and plant growth were evaluated. The experimental design was conducted as factorial arrangement with two GR soybean varieties, the Londrina (RR 59) and its near isogenic non-GM 59 Londrina called VAR 1; the second was Valiosa soybean (RR Conquista) and its near isogenic non-GM Conquista - Uberaba soybean called VAR 2. The plants were inoculated with arbuscular mycorrhiza fungi and rhizobia. The results showed that significant differences were observed among GM plants and their parental non-GM only for N biomass, AM colonization and cellulase activity. The presence of AM fungi had great influence on the functional groups of microorganisms while some enzymes activity decreased.

The relevance of a digestibility evaluation in the allergenicity risk assessment of novel proteins. Opinion of a joint initiative of COST action ImpARAS and COST action INFOGEST

The current allergenicity assessment of novel proteins is based on the EFSA GMO guidance. Recently, EFSA launched a new guidance document on allergenicity assessment of GM plants (2017). This document describes, amongst other topics, the new scientific and regulatory developments on in vitro protein digestibility tests. The EFSA GMO Panel stated that for in vitro protein digestibility tests, additional investigations are needed before any additional recommendation in the form of guidance can be provided. To this end, an interim phase is considered necessary to evaluate the revisions to the in vitro gastrointestinal digestion test, proposed by EFSA. This prompted the establishment of a joint workshop through two COST Action networks: COST Action ImpARAS and COST Acton INFOGEST. In 2017, a workshop was organised to discuss the relevance of digestion in allergenicity risk assessment and how to potentially improve the current methods and readouts. The outcome of the workshop is that there is no rationale for a clear readout that is predictive for allergenicity and we suggest to omit the digestion test from the allergenicity assessment strategy for now, and put an effort into filling the knowledge gaps as summarized in this paper first.

Annual Report on preparatory support for the statistical evaluation of the comparative assessment of GM plant field trials and for the evaluation of toxicological studies for GM plant food/feed safety – Lot 1: Comparative statistical analysis (2018)

Annual Report on preparatory support for the statistical evaluation of the comparative assessment of GM plant field trials and for the evaluation of toxicological studies for GM plant food/feed safety – Lot 1: Comparative statistical analysis (2018)

Annual Report on preparatory support for the statistical evaluation of the comparative assessment of GM plant field trials and for the evaluation of toxicological studies for GM plant food/feed safety – Lot 1: Comparative statistical analysis (2017)

Annual Report on preparatory support for the statistical evaluation of the comparative assessment of GM plant field trials and for the evaluation of toxicological studies for GM plant food/feed safety – Lot 1: Comparative statistical analysis (2017)

Annual Report of preparatory work on the toxicological studies and animal feeding studies performed under the EFSA contract OC/EFSA/GMO/2014/01, Lot 2 during the period 1/3/2017 to 27/11/2018

This report describes the tasks performed in the period 1/3/2017 to 27/11/2018 under the EFSA contract OC/EFSA/GMO/2014/01, Lot 2 on toxicological studies and animal feeding studies included in applications for market authorisation of genetically modified feed/plants under Regulation (EC) No 1829/2003. The tasks cover the check for study adherence to relevant EFSA guidance documents and to OECD Test Guideline no 407 (1995/2008), OECD Test Guideline no 408 (1998) and OECD Principles on Good Laboratory Practice. During the period covered by this report, preparatory work has been performed on six applications for GM plants submitted under Regulation (EC) No 1829/2003 for a total of three 28-day studies on newly expressed proteins and six 90-day studies in rodents on GM food/feed, using comprehensive checklist templates.

Розробка імуноензимного діагностикуму для виявлення гліфосат-резистентної генетично модифікованої сої

The goal of research was development of competitive ELISA kit for detection of genetically modified plants resistant to glyphosate. As the source for gene isolation we have used leaves of GM soybean RoundUp Ready, line 40-3-2 (Monsanto). Using polymerase chain reaction the gene encoding the enzyme 5-enolpiruvilshykimat-3-phosphate synthase (CP4 EPSPS) was isolated, which provides resistance to the herbicide glyphosate. Isolated gene has been cloned into plasmid vector pET24(a), creating genetic construct pET-CP4EPSPS. E.coli (BL 21(DE3)) has been transformed by genetic construct and over expressed after induction by 0.5 mM IPTG. Recombinant enzyme CP4 EPSPS with molecular weight 45 kD was purified as inclusion bodies and used as antigen for 96-well polystyrene plates immobilization. Monoclonal antibodies CP4 EPSPS mAb3 (Artron BioResearch Inc., Canada) was used for synthesis of horseradish peroxidase conjugate and receiving diagnostic reagent. We have shown that in the competitive ELISA we can detect at least 0.1% of GM plants resistant to glyphosate, on the example of soybean (line RR 40-3-2), moreover we can also detect rapeseed (line GT73) and sugar beet (line H7-1).

Genetic Engineering-Based Modern Approaches to Enhance Crop Resistance to Pests

Creation of genetically modified plants with an incorporated system protecting them from pests is one of the main goals in modern agricultural biotechnology. From the mid-1990s to the present, the majority of such transgenic crops is represented by carriers of the insecticidal Cry and Vip genes of the bacterium Bacillus thuringiensis. However, a tendency to change this strategy in favor of the incorporated defense systems based on RNA interference has become evident during the recent decade. Evolutional paths of phytophages' responses to insecticidal GM plants that are armed with these genetic constructions are discussed in the review.

Revisiting Risk Governance of GM Plants: The Need to Consider New and Emerging Gene-Editing Techniques.

New and emerging gene-editing techniques make it possible to target specific genes in species with greater speed and specificity than previously possible. Of major relevance for plant breeding, regulators and scientists are discussing how to regulate products developed using these gene-editing techniques. Such discussions include whether to adopt or adapt the current framework for GMO risk governance in evaluating the impacts of gene-edited plants, and derived products, on the environment, human and animal health and society. Product classification or definition is one of several aspects of the current framework being criticized. Further, knowledge gaps related to risk assessments of gene-edited organisms—for example of target and off-target effects of intervention in plant genomes—are also of concern. Resolving these and related aspects of the current framework will involve addressing many subjective, value-laden positions, for example how to specify protection goals through ecosystem service approaches. A process informed by responsible research and innovation practices, involving a broader community of people, organizations, experts, and interest groups, could help scientists, regulators, and other stakeholders address these complex, value-laden concerns related to gene-editing of plants with and for society.

RNA-guided multiplex genome engineering using cas9 nucleases for crop improvement: A review

Crop improvement through genome editing techniques has aroused substantial excitement among agricultural scientists as it offers opportunities for precise and predictable modification directly in elite cultivars or accessions which are not possible by time-consuming conventional breeding schemes. CRISPR-Cas9, a recently evolved gRNA-guided nuclease technology, is crucial in plant genome modifications which enable us in carrying out rapid and extremely precise mutations at pre-determined genomic sites by using guide RNA which is complementary to the target locus.This targeted DNA is then cleaved by Cas9 nuclease to generate double-strand breaks (DSBs), followed by subsequent DNA repair via non-homologous end joining (NHEJ) or homology-directed repair (HDR) mechanisms leading to the disruption of gene functions in plants. Multiplexed CRISPR-Cas9 exploits multiple gRNA sequences for simultaneous editing of multiple sites within the same genome, demonstrating its efficiency over traditional gene pyramiding. CRISPR/Cas9 is relatively convenient and easier to implement technology that has proven to be extremely efficient and versatile as compared to previously developed tools like ZFNs and TALENs and hence, it has been considered as a revolutionary technique in crop genome editing. The technique is continuously evolving to produce variants with its application in enhancing the agricultural performance of most crops. Moreover, plants developed by this technique have been considered superior to transgenic plants due to non-involvement of foreign DNA, hence, alleviating safety issues associated with GM plants. The current review briefly emphasizes on the potentiality of this game-changing technology in revolutionizing the basic plant genetic research and plant breeding.

Limen, Non-Toxic Recombinant Plant Defensin and Its Effect against Pathogenic Yeast and Fungi

Limen is a small cysteine-rich peptide that was originally isolated from lima beans, and its activity against Micromycetes was described. Recombinant expression of limen in both a prokaryotic and plant system was performed to demonstrate a wide selection of its activities with potential for further use. At first limen was expressed in E. coli cells and after purification steps, its antimicrobial activity was tested on pathogenic fungi. This showed that limen can significantly eliminate the growth of several pathogenic yeasts such as Candida albicans, while its hemolytic activity was not recognizable. To confirm limen’s activity in vivo, transformation of the barley Hordeum vulgare L. was chosen. Transgenic barley was obtained using Agrobacterium-mediated transformation and the production of limen peptide in plant tissues was verified by molecular techniques. Young barley plants were repeatedly exposed to Fusarium oxysporum spores. Increased resistance to the most important pathogen was followed using chlorophyll measurements as indicators of fusariosis. Focusing on the food safety aspect of the use of GM plants, cytotoxicity was followed in extracts from transgenic stressed plants. No or a lower cytotoxic effect was found on human keratinocytes and human embryonic kidney cells than extracts from the non-transformed plants.

EFSA Scientific Colloquium 24 – 'omics in risk assessment: state of the art and next steps

In recent years, the development of innovative tools in genomics, transcriptomics, proteomics and metabolomics (designated collectively as 'omics technologies) has opened up new possibilities for applications in scientific research and led to the availability of vast amounts of analytical data. The interpretation and integration of 'omics data can provide valuable information on the functional status of an organism and on the effect of external factors such as stressors. The European Food Safety Authority's (EFSA) 24th Scientific Colloquium on 'omics in risk assessment: state of the art and next steps explored the opportunities for integration of datasets produced via specific 'omics tools within the remit of EFSA's risk assessment approaches and tried to build further towards concrete paths of implementation. Discussions focused on genomics in microbial strain characterisation, metabolomics for the comparative assessment of GM plants and the use of 'omics for toxicological and environmental risk assessment. From the Colloquium it became clear that 'omics technologies are a valuable addition in some aspects of risk assessment of food and feed products and the environment, especially now that this technology is almost mature and stable. However, a consistent reporting framework for data collection, processing, interpretation, storage and curation should be further drawn up together with national and international organisations before 'omics technologies can be routinely used in risk assessment. For 'omics datasets in chemical and environmental risk assessments, the use of 'omics technologies alongside current toxicological or environmental risk assessment approaches is needed to re-inforce confidence and expertise before implementation of these datasets as a standalone tool in risk assessment. Test cases could be worked out to enhance confidence in the use of 'omics datasets in risk assessment.