According to the Law “On Seed Production”, all varieties or hybrids included in the state register shall be provided for genetic passports, as well as making a list of plants’ species, the production of which is aimed at ensuring food security. However, the use of morphological characteristics does not allow distinguishing genetically similar samples, identifying hidden genetic variability and ensuring control of the homogeneity of the initial material. In connection with these limitations, the purpose of the current work was to develop approaches for the selection of molecular genetic mic- rosatellite markers (SSR markers) to differentiate common wheat varieties. In accordance with the purpose, there has been conducted a literature review regarding methods for genetic certification and evaluation of the diversity of spring common wheat based on polymorphism of microsatellite loci, and there has been generated a set of Triticum aestivum genome assemblies from the NCBI Gen Bank. Based on the literature data, there have been selected the most polymorphic SSR markers using the invented algorithm. The analysis of databases of SSR markers in the genome of common wheat using bioinformatics methods allowed establishing a minimal discriminatory set of 20 markers that can detect 419 different alleles in Triticum aestivum. The results obtained can become the necessary theoretical foundation for further practical research.

The aim of the study was to study the state and reaction of cytoskeletal elements, microtubules and actin filaments, in root cells of Samsun tobacco plants and its transgenic line expressing the FeSOD1 gene from Arabidopsis thaliana with the pea rbcS leader sequence for the localization of the gene product in chloroplasts encoding Fe-dependent superoxide dismutase, constitutively inducing intracellular oxidative stress, by increasing the H2O2 pool for a long-term effect of moderate concentrations of NaCl and Na2SO4. The main hypothesis was to identify the positive protective effect of controlled constant oxidative stress on the stability of the most sensitive system that provides growth by division and growth by extension (the tubulin cytoskeleton) and effective intracellular transport and structural stability (the actin filament system). Localization of the microtubule cytoskeleton and actin filaments using antibodies to tubulin clone DM1α and actin clone 10-B3 by transmission electron microscopy and immunocytologically, detected by treatment with the second antibodies conjugated with Alexa‑488, made it possible to establish signs of reorganization and disassembly of the actin filament network under the action of NaCl and Na2SO4 as in control and in transgenic plants. At the same time, in transgenic plants, differences can be noted even without exposure, which indicates the effectiveness of this method for stimulating a protective response. These data suggest that the state of the system of the tubulin cytoskeleton and actin filaments may be an indicator of the resistance of FeSOD1 transgenic plants to salinity. A relationship has also been established between the reorganization of the cytoskeleton and vacuolization, especially with Na2SO4.

The creation of varieties resistant to different diseases and changing climate conditions, as well as high-quality berries, is the main goal of grape breeders. One of the approaches is the hybridization of cultivated Vitis vinifera L. varieties with wild and/or ancient accessions. A comprehensive study of the grapes’ genetic resources is highly necessary for a choosing of candidates for breeding process. The purposes of our study were: the determination of phylogenetic relationships between the 50 varieties and wild representatives of the Vitis vinifera L. species, compliance checking of the model of ecological and geographical groups to the varieties we have selected, and determination of the origin of some Crimean and other autochthonous grape varieties. The study was conducted on the basis of microsatellite DNA analysis for nine SSR markers approved by Vine and Wine (OIV) (VVS2, VVMD5, VVMD7, VVMD27, VVMD28, VVMD25, VVMD32, VrZAG62, VrZAG79) using the technology of genetic identification of grape varieties developed and published earlier. This technology includes multiplex PCR for all markers followed by the electrophoretic analysis of the obtained DNA fragments in one capillary of a genetic analyzer. For each studied sample, a genetic profile was obtained in the form of an individual set of lengths of DNA fragments, which allowed us to use the mathematical method of “Neighbor Joining” to construct a dendrogram reflecting phylogenetic relationships within our sample of varieties. As a result of our study, it was found that the model of ecological and geographical groups of ancient autochthonous varieties in the studied sample does not reflect the phylogenetic relationships between the samples. The spread of cultivated grape varieties at the initial stage of the development of viticulture went from east to west. The studied autochthonous Crimean grape varieties were introduced from other regions and do not originate from local wild forms of V. vinifera L.

A genotyping technology has been developed based on nine microsatellite loci (VVS2, VVMD5, VVMD7, VVMD27, VVMD28, VVMD25, VVMD32, VrZAG62, and VrZAG79). It can be used for efficient, accurate, and fast identification of varieties and forms of Vitis grapes. The proposed approach includes multiplex PCR of all loci followed by electrophoretic analysis of DNA fragments in one capillary of a genetic analyzer. The application of an additional length standard, an allelic ladder, consisting of all possible DNA fragments of the analyzed microsatellite loci, is one of the key features of the technology, which ensures the accuracy and reproducibility of the results. The advantage of the proposed technology is the possibility of standardizing and automating the procedure using 96-well plates, which opens the possibility of conducting mass analyses. As a result of the study of the varieties and forms of the Vitis genus species, genetic passports were created, according to which a dendrogram was constructed, reflecting the genetic relationship of the studied samples. The developed technology makes it possible to distinguish varieties and wild-growing forms of grapes; it can be used for their identification and determination of the genetic distance between them, as well as for assessment of planting material and protection of breeders’ rights.

Production of mesophilic protoplasts in potato (Solanum tuberosum L.) for transient gene expression is a mandatory technological stage in testing efficiency of new genetic constructs and CRISPR/Cas genome editing. In this study, isolation of protoplasts was carried out from leaves of potato plants of nine Russian cultivars grown in vitro for 6-7 weeks in vessels with foil caps, which prevented gas exchange with the environment. We first ever demonstrated the possibility of obtaining 2.4∙106 to 4.6∙106 viable protoplasts from one gram of aseptic plant leaves depending on the used potato cultivar. We established that the level of transfection of protoplasts with the genetic construct pHBT-sGFP-NosT amounts from 10 to 49% depending on the cultivar, which is sufficient for subsequent successful analysis of the efficiency of genetic constructs and potato genome editing. potato, Solanum tuberosum, protoplasts, plasmid DNA, transfection, transient expression, gfp The work was financially supported by the Comprehensive Research Program «Development of Potato Breeding and Seed Production».

The possibility of freeze drying of plant material and its grinding in a shaking bead mill to determine the activity of nitrate reductase (NR) was studied. The effectiveness of applying this approach to high throughput mass screening of crops was shown. The assay was carried out on seedlings of common wheat (Triticum aestivum) of the following cultivars: Altigo, Vassa, Grom, Doka, Soberbash, Starshina, Fisht and spring wheat Novosibirskaya 67. The crops were grown during 4-5 weeks on substrate without nitrogen and on substrate supplemented with 50 millimol / l (mM) of potassium nitrate. Nitrate reductase in plants retained its activity after lyophilization and grinding of dried leaves in a mill. The proposed protocol for NR activity assay is suitable for plant lysates with an NR activity suffi cient to form nitrite in the range of 5–120 micromoles / l (μM) in 800 μl of reaction mix (for instance, freeze-dried sample originated from 100 mg of wheat seedling leaves). Centrifugation of a plant lysate at 20,000 g almost did not change NR activity compared to 12,000 g that is achievable for most lab centrifuges. Lysates from fresh leaves contained signifi cantly more total protein than lysates from lyophilized leaves (with an equal amount of starting wet material). The difference in the nitrate-reducing activity in lysates from fresh and lyophilized leaves was not as high as the difference in protein concentration. Thus, the activity of NR calculated per g of total protein was higher in lyophilized leaves than in fresh leaves. The activity of NR was signifi cantly induced by nitrate for all cultivars. The basal and nitrate-induced NR activity varied widely between the cultivars, and the induction ranged from 2.5 fold for Novosibirskaya 67 variety and 2.7 fold for Vassa to 5.4 for Altigo and 5.7 fold for Grom.

РЕГУЛЯЦИЯ ОТТОКА АССИМИЛЯТОВ В ЗЕРНОВКИ ДЛЯ ПОВЫШЕНИЯ ПРОДУКТИВНОСТИ ПОСЕВОВ РИСА (Oryza sativa L.) ПРИ ВЫРАЩИВАНИИ У СЕВЕРНОЙ ГРАНИЦЫ АРЕАЛА КУЛЬТУРЫА.Х.ШЕУДЖЕН 1, 2 , Т.Н.БОНДАРЕВА 1, 2 , П

РЕГУЛЯЦИЯ ОТТОКА АССИМИЛЯТОВ В ЗЕРНОВКИ ДЛЯ ПОВЫШЕНИЯ ПРОДУКТИВНОСТИ ПОСЕВОВ РИСА (Oryza sativa L.) ПРИ ВЫРАЩИВАНИИ У СЕВЕРНОЙ ГРАНИЦЫ АРЕАЛА КУЛЬТУРЫА.Х.ШЕУДЖЕН 1, 2 , Т.Н.БОНДАРЕВА 1, 2 , П

Analysis of the moisture dynamics of leaves, stems, and panicles, as well as caryopsis filling, after treating rice crops with senicants showed that senication (if necessary) should be carried out for acceleration of ripening and biosynthesis of organic substances when the sum of temperatures above 15°C has reached 700–750°C. When carried out during this period, it does not cause premature dehydration of leaves and panicles and does not slow the caryopsis filling, which causes caryopsis moisture to reach 25% 3–10 days earlier and an increase of 3.5–10% by weight of panicles and rice yield by 0.30–0.39 t/ha in comparison with the control. It is viable to use aqueous solutions of ammonium nitrate at the rate of 15 kg/ha or carbamideammonium mixture CAM-32 diluted with water in a ratio of 1: 5 modified by adding 400 g/ha of manganese with a rate of working solution of 200–250 L/ha for this purpose.

The capacity of mesocotyls to agrobacterially transform four maize lines 4766, RB-179, L-1, L-2 and the conditions for their in vitro regeneration have been studied. Regeneration of maize mesocotyls was observed in media containing various auxins and cytokinins combinations. The efficiency of the process depended on the qualitative and quantitative compositions of growth regulators. The frequency of regenerated explants and the average number of shoots that form on one explant increased with increased cytokinin 6‑BAP and defoliant Dropp (thidiazuron) concentrations in the medium. 6-BAP (3–5 mg/L) and Dropp (3 mg/L) in combination with auxins (IAA, 0.5 mg/L and 2,4-D, 2 mg/L) were shown to be most effective for the mesocotyl regeneration. The number of regenerated explants reached 59.0%, whereas the average number of shoots per explant was 7.5 after 6 weeks of cultivation. The rooting and growing of regenerated shoots was successfully carried out on MS medium supplemented with 0.2 mg/L IBA. The capacity of the mesocotyl was determined by Agrobacterium tumefaciens–mediated transformation (ATMT) with an AGL0 strain containing the pVecActI-GUS genetic construction encoding the reporter gene uidA under the control of the promoter of rice actin ActI gene and the selective hptII hygromycin resistance gene. PCR analysis demonstrated that few regenerants with resistance to the selective agent contained the insertion of the hptII antibiotic-resistance gene in their genomes. Thus, it was shown that maize mesocotyls can be used as explants for agrobacterial transformation.

In the Russian Federation any food containing more than 0.9% DNA of genetically modified organisms (GMO) have to be specially labeled. The most widely used reference method for GMO detection is real-time polymerase chain reaction (real-time PCR). Two real-time PCR kits for identification genetically modified maize lines 5307 and MON89034 permitted for use in food and feed in the Russian Federation since 2014 were designed.

Исследовано влияние типа экспланта, а также вида и концентрации регуляторов роста на индукцию процессов каллусообразования и соматического органогенеза побегов in vitro четырех генотипов томата российской селекции. Проведено цитологическое изучение каллусной ткани. Установлено, что сорта томата обладают существенно большей способностью к непрямому органогенезу побегов по сравнению с гибридом F1. Для большинства исследуемых генотипов наибольшая частота соматического органогенеза побегов, а также их число на один эксплант были отмечены при культивировании семядольных листьев на питательной среде Мурасиге–Скуга, содержащей 2 мг/л зеатина в сочетании с 0.1 мг/л 3-индолилуксусной кислоты. Разработана эффективная методика непрямого соматического органогенеза побегов из различных эксплантов сортов томата с частотой >80%.

The influence of explant type as well as of the type of growth regulators and concentration on callus induction and somatic organogenesis of shoots was studied in vitro on four tomato genotypes of Russian breeding. Cytological study of callus tissue was conducted. It was established that tomato varieties have a substantially greater ability to indirect shoot organogenesis compared with the F1 hybrid. The highest frequency of somatic organogenesis of shoots, as well as their number per explant, was observed for most of the genotypes studied during the cultivation of cotyledons on Murashige-Skoog culture medium containing 2 mg/L of zeatin in combination with 0.1 mg/L of 3-indoleacetic acid. An effective protocol of indirect somatic organogenesis of shoots from different explants of tomato varieties with a frequency of more than 80% was developed.

During 1–7-day stages of ontogenesis (for example, in winter rye), biochemical changes in the aerial part of perennial grasses with adaptive Triticum-type proteins in the process of entering into the state of anabiosis were studied. It was shown that, at this point, biomass of the aerial part of plant was much higher than of the root system, and it accumulated raffinose, sucrose, glucose, and half-cystine. Synthesis of raffinose and accumulation of half-cystine continued until the stabilization of negative temperatures; as for sucrose and glucose, it ceased upon reaching the average temperature of the physiological zero, then the content of half-cystine and raffinose increased sharply, and the content of sucrose and glucose decreased at the same time. In the aerial parts of plants, fructose and phenylalanine were found, and their content increased upon reaching freezing temperatures. It was concluded that the polymerization of water-soluble proteins occurred in perennial grasses in a state of anabiosis. These proteins, along with raffinose, serve as cryoprotectants; sucrose and glucose are substrates for respiration; fructose is a substrate for respiration and protective metabolite; and phenylalanine is a substrate for the future synthesis of antioxidants.