Vol 20 (2022): Supplement

Chloroplasts provide life on our planet, by creating carbohydrates, amino acids, lipids and O₂ through the process of photosynthesis. Two billion years ago, a eukaryotic ancestor entered symbiosis with photosynthetic cyanobacterium, giving rise to chloroplasts. During evolution, chloroplasts transferred most protein-coding genes of free-living bacteria to the nucleus, but retained a semi-autonomous genetic status — their own genomes and transcription and translation machinery.

The genetics of plastids began in 1909, when Baur and Correns described the non-Mendelian, maternal inheritance of Pelargonium leaf color: green, white and variegated. The presence DNA in chloroplasts (chDNA) was prove in the 1960s, and later have been shown, that male chDNA in zygotes is specifically degraded, assuring maternal inheritance in most plants.

The plastid transformation was first described in the unicellular green alga Chlamydomonas reinhardtii [1]. This method of plant biotechnology is an alternative to traditional introduction of foreign DNA into the nucleus, and has a number of advantages: (a) the large copy number of chDNA offers high-level transgene expression; (b) site-specific integration of foreign genes reduces the number of transgenic lines to be evaluated; (c) the lack of gene silencing or position effects in chloroplast genomes; (d) the chloroplast employs a prokaryotic gene expression system, and allows the expression of polycistronic genes [2]. Thus, chloroplasts are becoming attractive hosts for the introduction of new agronomic traits, as well as for the biosynthesis of high-value pharmaceuticals, biomaterials and industrial enzymes.

Plants interact with a wide range of soil microorganisms, while interactions with symbiotic microorganisms are the most important for them. Symbiosis with nitrogen-fixing nodule bacteria provides a significant advantage in the existence of plants in nitrogen-poor soils. Since only plants from the Fabales and Rosales (a single representative Parasponia) enter into symbiosis with nodule bacteria, an idea about expanding the number of plants entering into such interactions became popular. To solve the problem of constructing new symbiotic systems, it is necessary to provide recognition of the symbiont (by transferring receptor genes into non-legume plants), morphogenesis of the nodule, and its infection.

We have managed to introduce the genes encoding receptors to surface components of rhizobia into the non-legume plants, and it provided increased colonization by nodule bacteria. It may improve the growth and development of non-legume plants and ensure their greater resistance to phytopathogens. Genome analysis of non-legume plant hop Humulus lupulus showed the existence of genes involved in symbiosis regulation, but some important regulators such as gene encoding NIN transcription factor were lost. Using genetic engineering approaches, the hop plants carrying the gene encoding NIN transcription factor were created. Analysis of these plants may provide important information about regulation of organogenesis in non-legume plants.

The work was financially supported by RSF 21-16-00106.

Genetic transformation of most dicotyledonous plants by Rhizobium rhizogenes (also known as Agrobacterium rhizogenes) results in production of composite plants plants consisting of wild-type shoot and transgenic root system. Composite plants are the suitable model for investigation of hormonal mechanisms related to development of the root system as regulatory links between the root system and the shoot maintains in such plants.

In most plants initiation of lateral root primordia occurs above the elongation zone [1]. However, in cucurbits and some other species, including important cereal crop buckwheat (Fagopyrum esculentum Moench), lateral root primordia initiation and development occurs in the apical meristem of the parental root [2, 3].

The phytohormone auxin is a key regulator of lateral root development. Fusions of auxin-responsive promoters and reporter genes can be used to study the role of auxin in the development of root system of non-model plants such as cucumber (Cucumis sativus L.) and buckwheat [4].

The “agrobacterium” — mediated transformation technique of cucurbits [5] has been adapted for buckwheat. R. rhizogenes strain R1000 was used in all transformations. Set of binary vectors based on pKGW-RR-MGW or pKGW-MGW was developed to study auxin response maxima (DR5::mNeonGreen) or auxin transport (fusions of genes encoding auxin efflux proteins PIN and mNeonGreen).

Pattern of auxin response maxima was similar in both species and included quiescent center and initial cells, columella, xylem cell files and lateral root primordia on all stages of development. Members of CsPIN1 (CsPINb and CsSoPIN1) group contributed unequally in generation of auxin maximum required for lateral root primordium initiation.

The research was supported by the RFBR grant 20-016-00233-a.

Radish (Raphanus sativus L.) is an agronomically important root crop belonging to the Brassicaceae family. A genetic collection of inbred radish lines, which was based on contrasting genetically determined traits including trait “ability to develop tumors”, was created at St. Petersburg State University in the middle of the 20^th century [1]. Full genetics network controlling this trait still remains unclear [2], and elucidation of its mechanisms can help reveal the key regulators of systemic mechanisms that control cell proliferation and differentiation.

The purpose of this work is to identify single nucleotide polymorphisms (SNPs) and InDels in genes which are candidates for participation in the tumor development process within in the tumor-forming radish line compared to the non-tumor radish line.

We have assembled the genomes of two lines contrasting in the ability to tumorigenesis, annotated them, aligned sequences per assembly, identified candidate genes and differences in the structure of these genes in contrasting radish lines using bioinformatics tools. Bioinformatics data were confirmed by the sequencing by Sanger method.

As a result, in the tumor-forming radish line we identified 151 genes with InDels in their coding regions, which led to various variants of frameshift. Moreover, we detected 39 genes with single nucleotide substitutions (SNPs). According to the gene pathway enrichment analysis, the corresponding genes were classified into the several groups.

Thus, the data obtained will allow us to clarify in more detail the genetic control mechanisms of the tumor development in radish.

The work was supported by the Ministry of Science and Higher Education of the Russian Federation in accordance with the agreement No. 075-15-2022-322.

Somatic embryogenesis (SE) is one of the plant regeneration pathways. This process is very similar to zygotic embryogenesis, but embryos develop not from zygote, but from vegetative tissues. This process is widely used in biotechnology for plant transformation and propagation. Somatic embryos can derive directly from the vegetative tissues or through the formation of callus. The search for SE stimulators is very important for plant biotechnology.

Several genes were reported to be the regulators of this process, among them the WOX (WUSCHEL-RELATED HOMEOBOX) family members are presented. These genes encode homeodomain-containing transcription factors, participating in different developmental processes. WOX2 is known for its participation in zygotic embryogenesis. It is expressed in the zygote and later in the apical domain of the embryo. We study the role of this gene in the somatic embryogenesis. Overexpression of MtWOX2 in some cases leads to the development of embryogenic calli with increased size. We performed transcriptome analysis of Medicago truncatula calli with overexpression of this gene compared to the calli overexpressing GUS.

It was shown that MtWOX2 overexpression led to the changes in expression levels of genes, enriched with several GO pathways, including groups related to oxidative stress and ROS formation, response to toxic substance and auxin-activated signaling pathway. Among differentially expressed genes there are members of several TF families, e.g. MADS-box, BHLH, MYB, bZIP and others. These genes may regulate embryogenic callus development. Together, these results can be used for the search of new morphogenic regulators applicable for plant transformation.

The research was supported by RFBR grant 20-016-00124.

Plant organisms are the objects of green biotechnology, which have been used by humans in various areas of life. The family of legumes (Fabaceae) that is being studied in this work is not an exception here. It is a very diverse family widespread throughout the globe.

Barrel medic (Medicago truncatula) and common pea (Pisum sativum), members of the legume family, were selected in this study for the development of an vital imaging system for transgenic tissues. As a part of the work, we tested an imaging system with post-mortem staining based on the GUS reporter, as well as a system for vital detection of transgenic tissue with fluorescent proteins GFP and DsRed. At this stage, vital imaging systems based on betalain and anthocyanin staining of transgenic tissues are designed and currently being adapted to the model objects under study.

The results of this study may be useful for subsequent attempts to solve the problem of the low efficiency of transformation of P. sativum. Moreover, such a system should be useful in the study of gene regulatory networks and factors that regulate the process of induction and subsequent development of somatic embryos. In addition, it can be used to track the dynamics of expression of genes of interest on living objects, while studying other fundamental and biotechnological processes.

The study is supported by the Russian Foundation for Basic Research (20-016-00124).

Pisum sativum L. (pea) is one of the most important agricultural crops, because its seeds have high protein content, and, due to its ability have symbiotic relationships with nitrogen-fixing bacteria, these plants need less fertilizers. Nevertheless, we are faced with the need to improve old and create new methods for obtaining novel varieties of peas and other agricultural plants. The formation of regenerated pea plants is difficult to achieve in the in vitro culture. Accordingly, transformation of this species is a laborious process. In this regard, the search for morphogenic regulators of somatic embryogenesis (SE) in pea is an urgent problem. A number of publications reported on the genes regulating the SE process in a model plant from the legume family, Medicago truncatula [1]. In our study, we search for the in vitro cultivation system in peas, suitable to test the effect of putative SE regulators in this species. We tested several pea transformation techniques using different explant variants: embryonic axes from mature and immature seeds, as well as shoot apexes. Out of the tested options, the transformation of mature seeds turned out to be optimal. We also designed a set of DNA constructs in silico, which are suitable for the search of morphogenic regulators in peas.

In modern pharmacology, there is a tendency to move from low molecular weight drugs to protein drugs. Interferon is among them. Interferon is used in animal husbandry to prevent and treat viral and bacterial diseases, as a single drug and as an adjuvant for vaccines and antibiotics to increase their efficiency and reduce the amount of use. After interferon therapy, meat, milk and eggs can be consumed without restrictions. Currently, interferon for veterinary purposes is obtained using genetically modified organisms (mainly bacteria). The limiting factor is the high cost of the active substance isolating and purifying, which is up to 80–90% of the product cost. Edible plants-producers can solve this problem. Plants have the lowest price per unit of biomass, are not infected by mammalian pathogens, and have an eukaryotic system of protein synthesis.

Transgenic tobacco plants (Nicotiana tabacum L.) for the production of bovine gamma-interferon were obtained by agrobacterium mediated transformation at the Department of Genetics and Biotechnology, Saint Petersburg State University. Testing of this model showed that interferon is successfully produced in plants and retains biological activity, including oral administration. Problems were also identified: tobacco toxicity and low levels of protein accumulation. At the next stage of the experiment, it was decided to transform edible carrot (Daucus carota) plants, modify the target protein (shorten) to increase its resistance to proteolytic degradation, and use the pSRD1 root-specific promoter. At present, the corresponding transgenic constructs have been obtained, the effectiveness of which will be determined first on the carrot hairy roots, and then on whole carrot plants.

The research was supported by the Ministry of Science and Higher Education of the Russian Federation in accordance with agreement No. 075-15-2020-922 date 16.11.2020 on providing a grant in the form of subsidies from the Federal budget of Russian Federation. The grant was provided for state support for the creation and development of a World-class Scientific Center “Future Agrotechnologies”.

Agrobacterium tumefaciens is unique in that it can facilitate the interkingdom transfer of DNA and genetically modify its plant host. While Agrobacterium has been coopted for use in the genetic modification of plants, it is also a major pathogen, causing crown gall disease in the nursery, orchard, and vineyard industries. Pathogenicity in Agrobacterium is the result of two components. First is the Ti plasmid, which carries virulence genes and the transferred T-DNA region. The second component is the chromosome of Agrobacterium, which comprises diverse bacterial lineages and multiple species-level groups. The Ti plasmid can be transferred from strain to strain, diversifying the pathogen and complicating efforts to understand its epidemiology. This system provides an opportunity to study transmission of plasmids and their impact on disease persistence and spread. However, the movement of plasmids, and diversity of chromosomal lineages, means that conventional methods of using whole genome SNPs to track outbreaks are not sufficient, and new techniques must be developed. Additionally, Ti plasmids, like Agrobacterium, are genetically diverse and represent multiple plasmid types. Using a framework of >200 sequenced Agrobacterium genomes isolated from around the world, and a previously developed model of Ti plasmid types, we modelled their epidemiology. Key to this study was that we first separately analyzed plasmids and strain. Combining results revealed links between nurseries, potential horizontal transfer of the plasmid between strains within nurseries, global spread of plasmids, and long-term persistence of plasmids in the agricultural system. Agricultural practices have the potential to promote the diversification of pathogens and the emergence of new pathogen lineages.

Soil bacteria “Agrobacterium” are able to transfer fragments of their plasmids, so-called T-DNA, into plants. T-DNA integrated into plant’s genome is called cellular T-DNA (cT-DNA) [1]. Plants transformed in nature are considered natural genetically modified organisms (nGMOs). For the first time, nGMOs were described within the genus Nicotiana. To date, more than 50 nGM species are known [2, 3], among which nGMO in the genus Nicotiana are the most well studied. Within this genus 3 subgenera are distinguished, those are Tabacum, Petunioides, and Rustica. CT-DNAs in natural genetically modified representatives of the subgenus Tabacum are studied in detail [4, 5]. We know how many cT-DNA those species carry as well as the composition of the cT-DNA, which allows us to propose scenarios for the acquisition of cT-DNA by these species during their evolution. Species N. noctiflora and N. glauca belong to the subgenus Petunioides and they are not so well studied. We sequenced and assembled the genomes of N. noctiflora and N. glauca, to analyze their cT-DNAs. In the N. glauca genome we confirmed the presence of one cT-DNA, gT, discovered in 1983, and showed no other inserts. In the genome of N. noctiflora 2 cT-DNAs of different composition were found, NnT-DNA1 and NnT-DNA2. The data suggest a single agrotransformation act in the evolution of the species N. glauca, while the species N. noctiflora was transformed several times. Further study of cT-DNA in Nicotiana representatives belonging to different evolutionary branches of the genus will help to clarify the evolutionary history of the genus Nicotiana. In addition, the identification of changes that have occurred in the cT-DNA since its entry into the plant genome will help to elucidate the processes that occur with transgenes in plant genomes over long time intervals.

The work was supported by the Russian Science Foundation (21-14-00050) and was conducted using the equipment of the Research Center of St. Petersburg State University “BioBank”. The authors are grateful to professor Leon Otten, as well as Pavel Dobrynin, Dmitry Polev, Nikolai Ivanov, and Nicholas Sierro.

A few dozen naturally transgenic plants have been described to date [1]. Many of them contain DNA-sequences homologous to opine biosynthesis genes. Chemically, opines fall into two major structural classes: secondary amine derivatives and sugar-phosphodiesters [2]. The concentration of these molecules in the crude plant extract of nGMOs could be less than 1 pmol, which makes it difficult to study them. However, structure clarification of plant opines reveal their function in plants. The aim of our work is the development of protocol for large scale production, purification and definition of the chemical structure of plant opines.

The approach includes amplification of full-length opine synthase gene by PCR and cloning it using pENTR™/D-TOPO™ Cloning Kit (Thermo Fisher) according to its manual; subcloning in pDest527 using LR Clonase™ II Plus enzyme (Thermo Fisher) according to its manual; transformation NiCo21(DE3) chemically competent E. coli cells with obtained recombinant plasmids. After the confirmation of transgenic nature, the E. coli cells should be grown on medium with ampicillin 100 mg/l to an optical density at 600 nm 0.5–0.6. Then an equal volume of LB medium with 1 mM isopropylthio-β-galactoside (IPTG) should be added for induction opine synthesis. In case of synthesis of sugar-phosphodiesters, cultural media should also contain 0.02 M arabinose, glucose, and sucrose, respectively. Cells are cultivated during 4 hours for induction of opine synthesis. After that cells are pelleted by centrifugation for 10 minutes at 5000 rpm/min. The original strain and cells grown on the medium without IPTG are used as controls.

The cells are extracted with 80% methanol. The culture liquids are evaporated and redissolved in 80% methanol. The opines are separated by normal-phase chromatography using a CHROMABOND® Flash BT. Thin-layer chromatography is used to analyze the fractions, and to confirm the component opines-like compounds. High-resolution mass spectra are recorded on a Bruker micrOTOF 10223 mass spectrometer (electrospray ionization), eluent 80% MeOH. The ³¹P NMR spectra are acquired on a Bruker Avance 400 spectrometer (400, 100, and 162 MHz, respectively). The ¹H spectra are analyzed in D₂O, with residual solvent signals (7.26 ppm for ¹H nuclei) as the internal reference. The ³¹P NMR is measured relative to H₃PO₄.

The one phosphoric acid residue in the opine structure was confirmed by NMR spectroscopy.

We applied this method to characterize agrocinopine A-like compound in Nicotiana and propose it to produce opine-like plant compounds for further biochemical analysis.

This research was funded the Ministry of Science and Higher Education of the Russian Federation in accordance with the agreement No. 075-15-2022-322.

This work was partially carried out using equipments of the Saint Petersburg State University “Chemical Analysis and Materials Research Centre”.

Agrobacterium mediated transformation is one of the most studied examples of horizontal gene transfer between pro- and eukaryotes. During this process a part of the Ti-plasmid — T-DNA — is transferred into the plant cell genome. These sequences could be preserved in the genomes during evolution and inherited in a series of sexual generations. Such plants are described within the genus Vaccinium L. [1]. Our research team is currently analyzing natural transgenes in V. oxycoccos L., V. japonicum Miq., V. conchophyllum Rehder, V. emarginatum Hayata, V. myrtilloides Michx., V. virgatum Ait., V. corymbosum L., V. darrowii Camp, V. smallii A. Gray, V. praestans Lamb., V. ovalifolium Sm., V. myrtillus L., V. uliginosum L., V. vitis-idaea L.

Previously, analyzing the natural transgenes in another genus (Camellia L.) [2], we showed the importance of reconstructing the allelic states of transgenes for phylogenetic studies.

In this paper, we present a comprehensive approach for studying the allelic state of the rolB/C-like gene in plants of the genus Vaccinium. It combines molecular genetic and bioinformatic research methods.

Molecular genetic methods involve Sanger sequencing of a gene sequence in a large number of samples, while for each sample the sequence is presented as a set of polymorphic positions in binary form. Allele resolution occurs based on the description of alleles in homozygotes and a series of “subtractions” of known alleles in heterozygous samples. The second method involves the analysis of SRA (Sequence Read Archive) sequences available in the databases. SRA is a repository of high-throughput sequencing raw data.

Based on our work, we can conclude that both of these approaches make it possible to describe the allelic state of the rolB/C-like gene in representatives of the genus Vaccinium.

The work was performed using the equipment of the Resource Center of Saint Petersburg State University “Development of Molecular and Cellular Technologies” with the support of the Ministry of Science and Higher Education of the Russian Federation in accordance with agreement No. 075-15-2022-322 dated 04/22/2022 on the provision of a grant in the form of a subsidy from the Federal budget of the Russian Federation. The grant was provided as part of the state support for the creation and development of the world-class Scientific Center “Agrotechnologies for the Future”.

Naturally transgenic plants represent the result of Agrobacterium-mediated gene transfer. T-DNA of soil bacteria Agrobacterium integrated into plant’s genome is called cellular T-DNA (cT-DNA) [1]. Today, more than 50 species of naturally transgenic plants, or natural GMO (nGMO) are known [2, 3]. The function of cT-DNA in plants remains unknown. It is assumed that the fixation of transgenes could give plants different selective advantages depending on which genes had been integrated into the plant [4]. In order to clarify this issue, it is necessary to study more naturally transgenic plants. Until recently, the list of nGM plants contained less than 2 dozen species, but a search through genomic and transriptomic sequencing data made it possible to more than double this list [2]. In this work, we used the same approach, looking for cT-DNA genes in whole genome sequencing data that have appeared in the NCBI WGS since 2021. We found 14 new species of naturally transgenic plants, among which the most extended cT-DNAs were found in Triadica sebifera, Lonicera japonica, and Lonicera maackii. The cT-DNAs in these species are organized as imperfect inverted repeats. In the genomes of the species Paulownia fortunei, Apocynum venetum, Elaeagnus angustifolia, Erythroxylum havanense, E. densum, E. daphnites, E. cataractarum, Ceriops decandra, Camellia oleifera, Silene uniflora, short cT-DNAs containing only opine synthesis genes were found. We also estimated the approximate age of the cT-DNAs. The first described examples date back to the Late Paleogene, and the process continues to the present. Thus, we can conclude that natural GMOs are a widespread phenomenon, many aspects of which remain unclear, requiring additional research on the topic.

The article was made with support of the Ministry of Science and Higher Education of the Russian Federation in accordance with agreement No. 075-15-2022-322 dated 22.04.2022 on providing a grant in the form of subsidies from the federal budget of the Russian Federation. The grant was provided for state support for the creation and development of a world-class scientific center, “Agrotechnologies for the Future”.

Methylotrophic yeast Komagataella рhaffii (also known as Pichia pastoris) is widely applied in biotechnology for recombinant protein production. K. рhaffii particularly proved to be a successful host system for the synthesis of immunomodulators such as interferons [1].

In this study, we engineered K. рhaffii strains capable of producing the immunomodulatory protein Neoleukin (Neo-2/15). Neo-2/15 is an interleukin-2 mimetic, designed by in silico methods [2]. In preclinical studies on murine cancer models, Neo-2/15 showed superior therapeutic effect to inerleukin-2 with reduced toxicity.

In this work, we show that K. рhaffii can successfully synthesize and secrete Neo-2/15. We have obtained a number of K. phaffii strains, including Mut^S and Mut⁺, with different Neo-2/15 expression cassettes integrated into the genome, carrying up to five copies of Neo-2/15 gene. In fact, the higher number of Neo-2/15 gene copies in K. рhaffii genome allowed a higher protein yield.

In this study, we further developed a split marker approach [3] for yeast transformation using two DNA fragments, comprising of the expression cassette and the overlapping fragments of the marker gene. This allowed us to generate Mut^S strains with two copies using pPICZαB vector, which is not originally intended for Mut^S strain generation.

As a result, we demonstrated that K. phaffii is a perspective producer of Neo-2/15, providing wide opportunities to increase the production of this therapeutic protein.

In the modern world, due to the overconsumption of sugar-containing products, the problem of obesity is relevant. Among the many sweeteners that minimize sugar intake, a group of sweet-tasting proteins is up-and-coming. Brazzein is the smallest of the sweet proteins (54 aa, 6473 Da), and it is also safe for obese and diabetic people since it does not affect blood sugar and insulin levels. Brazzein has high thermal stability over a wide pH range: from 2 to 8 [1]. To increase the level of sweetness of brazzein, mutant variants of this protein were created through site-directed mutagenesis, the sweetest of which is triple mutant H31R/E36D/E41A, which is 22,500 times sweeter than sucrose [2]. Since the content of brazzein in the fruits of the natural source (Pentadiplandra brazzeana) is extremely low (0.2%), various methods have been developed to obtain brazzein using heterologous expression systems, which used as producers: bacteria (Escherichia coli, Lactococcus lactis), yeast (Pichia pastoris, Kluyveromyces lactis, Saccharomyces cerevisiae), plants (Zea mays, Oryza sativa, Lactuca sativa, Nicotiana tabacum) and animals (Mus musculus) [3–5]. Despite the short peptide sequence, the industrial production of recombinant protein faced several problems, including low protein yield (e.g in mouse milk it was detectable on western blot analysis only) and loss of sweetness. Аn extremely relevant and promising way to obtain recombinant brazzein is the optimization of extracellular expression in bakers yeasts with the GRAS (Generally recognized as safe) status, since the safety of these microorganisms for human health can potentially significantly reduce the number of brazzein purification steps and thereby reduce its cost to consumers.

Amyloids are fibrous protein structures often found in patients with severe diseases, such as Alzheimer’s, Parkinson’s diseases etc. A number of studies have shown that the production of heterologous amyloidogenic proteins in Saccharomyces cerevisiae strains results in formation of amyloid aggregates with properties similar to those found in mammals.

Amyloid aggregates formed in yeasts usually do not have their own phenotypic manifestation. To assess amyloidogenic potential of individual proteins a yeast test-system was developed under supervision of Prof. Y.O. Chernoff. The system is based on usage of genetically modified S. cerevisiae cells auxotrophic for certain growth factors, allowing effective phenotypic selection to search for amyloidogenic proteins within proteomes of various organisms [1]. Using this test-system, our laboratory evaluated amyloid potential of a spectrum of human proteins, the amyloidogenicity of which was previously predicted by bioinformatics algorithms. The proteins that have shown amyloidogenic potential in yeast-based model are being currently tested in vitro and in vivo. Some mutant Escherichia coli strains can be applied for studying propensity of heterologous proteins to form amyloids in vitro. Thus, application of genetically modified microorganisms makes it possible to identify new human amyloidogenic proteins and to improve predictive ability of bioinformatics algorithms.

The research is supported by RSF grant №20-14-00148 and by St. Petersburg State University (project No. 93025998). Authors acknowledge SPbSU Resource Centers “Chromas”, “Molecular and Cell Technologies” and “Biobank”.

The paper covers major threats associated with wide-range introduction and cultivation of transgenic plants due to germplasm mixing with that of indigenous species of natural plant communities and risks of transgenic plants’ adverse impact on the environment. Among them are: influencing non-target species, invasive power, possibility of GMP’s escaping into the environment by horizontal gene transfer as well as harmful effect on the soil biota.

Currently, herbicide- and pest-resistant genetically modified plants (GMP) became an integral part of contemporary agrotechnologies in many economies [1]. However, most countries lack national strategy providing science-based substantiated procedure of creating, distribution and safe production of GMP. Rapid development of agricultural biotechnology and GMP production offered many economical benefits but also caused concern due to their potential environmental impact. To date, truly negative effects of GMP production, revealed in the course of growing, are known: harmful effect of entomocide Cry-proteins (Bt endotoxins) on non-target biota, target phytophage resistance to insecticidal plants, phytophage species succession to replace the species eliminated in the agrocoenosis. Vertical transfer of GMP transgenes (repollination between transgenic plants and wild species or isogenic varieties), as well as slow decomposition of transgenic plants’ remains — all these factors can have remote environmental consequences [2, 3]. Wind-dispersed pollen of insecticidal GMP contaminates soil and open water reservoirs by toxins, thus posing potential hazards for aquatic organisms and geobionts (including rhizospheric organisms).

Thus, uncontrolled GMP production and introduction, creates a real threat of losing biodiversity and genetic diversity of indigenous plants due to “biological contamination”. Therefore, GMP cultivation and monitoring in the fields is of exceptional importance and must be regulated by a science-based national strategy. This strategy would allow to exclude agroecological and environmental genetic risks, to keep the genetic diversity of natural plant communities.

The development of genomic editing technologies has significantly intesificated the shortcomings of the legal regulation for genetic technologies as in general and as for new technologies of genetic engineering in particular. Because of imperfected legislation for genetic engineering, and in some cases because of its archaism, new genomic technologies, such as genetic editing technologies, which are growth drivers in science, could not be drivers in the economics. The outdated terms and concepts framework and the legal indetermination of using the products based on new genetic technologies (including genome editing) are barriers to achieving the goals to ensure the technological independence of Russia.

The basis of the current legal regulation system of genetic engineering in Russia is a focus to the process and technologies, i.e. methods of the product producing. This means that the changes within genetic information is not important, but only the method of development is. Such way of regulation provides a priori chronic lag in legislation, which could finally lead to technological lag as whole country. The transition to a product-oriented system, when the analysis of base of genetic information changes makes possible to clearly answer to the question of the methods used for this, will allow us to avoid the shortcomings within development of synthetic biology methods indicated earlier, as now as in the future.