Vaccination is the most simple and reliable approach of protection to virus infections. The most effective agents are live vaccines, usually low-virulence organisms for humans and closely related to pathogenic viruses or attenuated as a result of mutations/deletions in the genome of pathogenic virus. Smallpox vaccination with live vaccinia virus (VACV) closely related to smallpox virus played a key role in the success of the global smallpox eradication program carried out under the World Health Organization auspices. As a result of the WHO decision as of 1980 to stop smallpox vaccination, humankind has lost immunity not only to smallpox, but also to other zoonotic, orthopoxviruscaused human infections. This new situation allows orthopoxviruses to circulate in the human population and, as a consequence, to alter several established concepts of the ecology and range of sensitive hosts for various orthopoxvirus species. Classic VACV-based live vaccine for vaccination against orthopoxvirus infections is out of the question, because it can cause severe side effects. Therefore, the development of new safe vaccines against orthopoxviral infections of humans and animals is an important problem. VACV attenuation by modern approaches carried out by targeted inactivation of certain virus genes and usually leads to a decrease in the effectiveness of VACV in vivo propagation. As a result, it can cause a diminishing of the immune response after administration of attenuated virus to patients at standard doses. The gene for thymidine kinase is frequently used for insertion/inactivation of foreign genes and it causes virus attenuation. In this research, the effect of the introduction of two point mutations into the A34R gene of attenuated strain LIVP-GFP (ТК–), which increase the yield of extracellular enveloped virions (EEV), on the pathogenicity and immunogenicity of VACV LIVP-GFP-A34R administered intranasally to laboratory mice were studied. It was shown that increase in EEV production by recombinant strain VACV LIVP-GFP-A34R does not change the attenuated phenotype characteristic of the parental strain LIVP-GFP, but causes a significantly larger production of VACV-specific antibodies.

Cotyledon and leaf explants of two spring rapeseed varieties were transformed with Agrobacterium tumefaciens harboring a genetic construct with the gfp marker gene. In order to reduce the proportion of hyperhydrated shoots, which appeared during regenerant formation, we optimized sucrose content in the regeneration media. Analysis of the progeny obtained from T0 regenerants showed that in a number of lines the distribution of the gfp marker did not follow Mendelian segregation of a monogenic trait in self-pollinated plants, while in the progeny of the other lines of transgenic plants, the gfp marker was completely absent, although its presence had been confirmed in all selected T0 plants. We also found that in individual transformants gfp is randomly inherited throughout the central peduncle; its presence in the genome of seedlings does not depend on the location of the pod. Thus, both transformed and non-transformed cells were involved in the formation of gametes in T0 plants. In addition, marker segregation was different in plants of the T1 line obtained by nodal cuttings of a primary transformant, depending on the location of the cuttings on the stem of the original plant, indicating that the nature of T1 generation plants was also chimeric. Furthermore, we showed that propagation of plants by cutting followed by propagation by seeds formed as a result of self-pollination led to an increase in the proportion of transgenic plants in subsequent generations. The results obtained during the course of this study show that the transformants were chimeric, i. e. their tissues contained both transgenic and non-transgenic cells, and this chimeric nature was passed on to subsequent generations. We found that, in addition to nutrient media composition, other factors such as plant genotype and explant type also contribute to the rising of chimeric plants during transformation. Based on these results, we developed a simplified method, which consists of several rounds of a combination of cutting, seed production by self-pollination, and subsequent culling of wild-type plants, which significantly enriched descendent populations of the original rapeseed transformants with plants transgenic for the gfp marker.

The method of biological ballistics (biolistic transformation, genetic bombardment) of plants is one of the most modern methods used for direct gene transfer into plant cells. The main advantages of this method include the ability to simultaneously incorporate several target genes into the plant genome, carry out transfer without unnecessary agrobacterial parts and plasmid DNA sequences, and the short time needed to produce transgenic cells. For different plant objects, the efficiency of obtaining transgenic plants by the ballistic method varies from 1 to 3 %. For potato plants, the transformation efficiency is quite low at the moment and the selection of optimal conditions for biolistics is one of the pressing issues of practical biotechnology. This article presents a successful experience of introducing two genes of interest into two potato varieties using the biolistic approach. The results of biolistic transformation experiments are presented for two types of explants: potato internodes and calli of the varieties Aksor and Nevskiy. Of the 862 explants used for transformation, 56 regenerated plants were obtained. PCR screening of transformants revealed one plant with the insertion of the chitinase gene, one with the insertion of the endo-β-1,3-glucanase gene, and co-transformation by both genes was confirmed in four regenerants. The average transformation efficiency for potato explants was 0.7 %. A high number of regenerants (56) as opposed to a low number of transformants (6) reflects an attempt to increase the number of regenerants by using a lower concentration of the selective agent (kanamycin). Although this approach requires more effort, it can be used to produce potato lines with integrated genes of interest for further use in crop breeding. The lines of potato obtained in the current study by introducing two genes associated with the plant response to fungal pathogens will be further assessed for their resistance to fungal diseases and, if successful, will be used in potato crop breeding.

Phytophthora infestans Mont. de Bary is the main oomycete pathogen of cultivated crops in the family Solanaceae, especially potato (Solanum tuberosum). Because potato is the fourth most cultivated crop worldwide, its annual losses from late blight are tremendous. Studies of the basic mechanisms of interaction between potato and the late blight pathogen not only expand the fundamental knowledge in this area, but also open up new possibilities for regulating these interactions in order to increase resistance to the pathogen. The interaction of potato and the late blight pathogen can be considered from a genetic point of view, and it is interesting to consider both the response of the potato to the colonization process by P. infestans and the change in gene activity in late blight during plant infection. We can also investigate this process by changing the profile of secondary metabolites of the host and the pathogen. In addition to fundamental work in this area, applied work in the form of the development of new preparations for protecting potatoes is of no less importance. This review briefly describes the main stages of studies of potato resistance to late blight, starting almost from the first works. Much attention is paid to key works on changing the profile of secondary metabolites phytoalexins. A separate section is devoted to the description of both qualitative and quantitative characteristics of potato resistance to the late blight pathogen: their contribution to overall resistance, gene mapping, and regulation capabilities. Both types of traits are important for potato breeding: quantitative resistance due to R-genes is quickly overcome by the pathogen, while quantitative trait loci make it possible to create varieties with almost absolute resistance due to the pyramid of effective genes. The latest approaches in molecular biology make it possible to study translatomic profiles, which makes it possible to look at the interaction of potatoes and the late blight pathogen at a different angle. It has been shown that the process of potato colonization affects not only the activity of various genes and the profile of secondary metabolites: proteins­markers of the response to infection from potatoes have also been identified: they are pathogen-bound proteins and plastid carbonic anhydrase. On the part of P. infestans, fungal cellulose synthase proteins and haustorium-specific membrane protein were markers of infection. Thus, the review contains information on the most relevant complex studies of the genetic mechanisms of potato resistance to late blight.

About one-third of the world’s barley crop is used for malt production to meet the needs of the brewing industry. In this regard, the study of the genetic basis of malting quality traits and the breeding of malting barley varieties that are adaptive to their growing conditions are relevant throughout the world, particularly in the Russian Federation, where the cultivation and use of foreign malting varieties of barley prevails. The main parameters of malting quality (artificially germinated and dried barley grains) are malt extract, diastatic power, Kolbach index, viscosity, grain protein, wort β-glucan, free amino nitrogen, and soluble protein content. Most of these components are under the control of quantitative trait loci (QTLs) and are affected by environmental conditions, which complicates their study and precise localization. In addition, the phenotypic assessment of malting quality traits requires elaborate, expensive phenotypic analyses. Currently, there are more than 200 QTLs associated with malting parameters, which were identified using biparental mapping populations. Molecular markers are widely used both for mapping QTL loci responsible for malting quality traits and for performing marker-assisted selection (MAS), which, in combination with conventional breeding, makes it possible to create effective strategies aimed at accelerating the process of obtaining new promising genotypes. Nevertheless, the MAS of malting quality traits faces a series of difficulties, such as the low accuracy of localization of QTLs, their ineffectiveness when transferred to another genetic background, and linkage with undesirable traits, which makes it necessary to validate QTLs and the molecular markers linked to them. This review presents the results of studies that used MAS to improve the malting quality of barley, and it also considers studies that searched for associations between genotype and phenotype, carried out using GWAS (genome-wide association study) approaches based on the latest achievements of high-throughput genotyping (diversity array technology (DArT) and single-nucleotide polymorphism markers (SNPs)).

Among the natural pigments, anthocyanins are assumed to represent one of the most studied groups. Starting with the first studies on the physicochemical properties of anthocyanins carried out in the 17th century by British naturalist Robert Boyle, the science about these unique compounds has progressed substantially. To date, the structure and functions of anthocyanins in plant cells have been well studied, and the pathway of their biosynthesis is one of the most fully characterized pathways of secondary metabolite biosynthesis at both the biochemical and genetic levels. Along with these fundamental achievements, we are beginning to realize the potential of anthocyanins as compounds of industrial importance, as pigments themselves, as well as components of functional food that contribute to the prevention and reduction of risk of chronic diseases. For a long time, the biological activity of anthocyanins has been underestimated, in particular, due to the data on their low bioavailability. However, studies showed that in humans and animals, these compounds are actively metabolized and the bioavailability, estimated taking into account their metabolites, exceeded 12 %. It has been experimentally shown that anthocyanins have antioxidant, anti-inflammatory, hypoglycemic, antimutagenic, antidiabetic, anti-cancer, neuroprotective properties, and they are beneficial for eye health. However, the studies conducted cannot always explain the molecular mechanism of action of anthocyanins in the human body. According to some reports, the observed effects are not due to the action of anthocyanins themselves, but to their metabolites, which can be more biologically active because of their increased bioavailability. Other data ascribe the positive effect on human health not to individual anthocyanins, but to the whole complex of polyphenolic compounds consumed. The review summarizes the results of the studies of anthocyanins as components of functional food. Special attention is paid to genetic control of the pigment synthesis. These data are of particular importance in respect to the initiated breeding programs aimed at increasing the content of anthocyanins in cultural plants.

Changes in the environment force populations of organisms to adapt to new conditions, either through phenotypic plasticity or through genetic or epigenetic changes. Signatures of selection, such as specific changes in the frequency of alleles and haplotypes, as well as the reduction or increase in genetic diversity, help to identify changes in the cattle genome in response to natural and artificial selection, as well as loci and genetic variants directly affecting adaptive and economically important traits. Advances in genetics and biotechnology enable a rapid transfer of unique genetic variants that have originated in local cattle breeds in the process of adaptation to local environments into the genomes of cosmopolitan high-performance breeds, in order to preserve their outstanding performance in new environments. It is also possible to use genomic selection approach to increase the frequency of already present adaptive alleles in cosmopolitan breeds. The review examines recent work on the origin and evolution of Turano-Mongolian cattle breeds, adaptation of Turano-Mongolian cattle to extreme environments, and summarizes available information on potential candidate genes for climate adaptation of Turano-Mongolian breeds, including cold resistance genes, immune response genes, and high-altitude adaptation genes. The authors conclude that the current literature data do not provide preference to one of the two possible scenarios of Turano-Mongolian breed origins: as a result of the domestication of a wild aurochs at East Asia or as a result of the migration of taurine proto-population from the Middle East. Turano-Mongolian breeds show a high degree of adaptation to extreme climatic conditions (cold, heat, lack of oxygen in the highlands) and parasites (mosquitoes, ticks, bacterial and viral infections). As a result of high-density genotyping and sequencing of genomes and transcriptomes, prospective candidate genes and genetic variants involved in adaptation to environmental factors have recently been identified.

Mezenskaya horse (Mezenka) is Russia’s aboriginal breed. It is a domestic selection in the northern territories of Arkhangelsk region. The breed is perfectly adapted to the conditions of the Far North, and has a number of valuable economic and biological qualities. At present, it has a limited gene pool and is bred only in the Mezensky district, where one gene pool-breeding farm is operating and so is a number of basic farms, where selection and breeding activities take place with the breed. Due to a small population of Mezen horses, the challenge of preserving its intra-breed diversity is very urgent. To determine the degree of genetic variability in the Mezen population, the alleles-fond was monitored. A comparative analysis of the genetic structure of the breed was done on DNA microsatellites at time-intervals of 10 years (2000, 2010 and 2020). Crista samples of 198 horses were studied in specialized laboratories. It was established that the breed has wide genetic diversity in 17 loci of nuclear DNA. The population’s alleles-fond includes from 128, 139, and 133 alleles respectively (with an average value of 7.53, 8.18, and 7.82 alleles per locus). The most common alleles are AHT4O, AHT5N, ASB2K, ASB23S, CA425N, HMS1J, HMS1M, HMS2H, HMS3M, HMS7L, HTG4M, HTG6O, HTG7K, HTG7O and LEX3M. Mezen horses revealed 6 rare, lowfrequency (0.004–0.056) alleles not found in the horse populations of domestic selection. The average value of the polymorphic level (Ae) in the breed over the years is 4.16, 4.21 and 4.06, respectively. The highest polymorphism is found in locus ASB17 (6.49–6.90–6.76); the lowest, in locus HTG6 (1.71–1.66–1.67) and HMS7 (1.77–1.95–1.77). A slight deficit of heterozygous genotypes (Fis = 0.003) was observed in Mezen horses in 2010. In 2000 and 2020, the observed heterozygosity (Ho) exceeds the expected value (He), which indicates the absence of intra-population inbreeding (Fis = –0.014 and –0.011, respectively). The results obtained testify to the effectiveness of breeding activities carried out to preserve, improve and maintain genetic diversity in the population.

Individual behavioral differences are due to an interaction of the genotype and the environment. Phenotypic manifestation of aggressive behavior depends on the coordinated expression of gene ensembles. Nonetheless, the identification of these genes and of combinations of their mutual influence on expression remains a difficult task. Using animal models of aggressive behavior (gray rats that were selected for a reaction to humans; tame and aggressive rat strains), we evaluated the expression of 10 genes potentially associated with aggressiveness according to the literature: Cacna1b, Cacna2d3, Drd2, Egr1, Gad2, Gria2, Mapk1, Nos1, Pomc, and Syn1. To identify the genes most important for the manifestation of aggressiveness, we analyzed the expression of these genes in two generations of rats: 88th and 90th. Assessment of gene expression levels was carried out by real-time PCR in the hypothalamus of tame and aggressive rats. This analysis confirmed that 4 out of the 10 genes differ in expression levels between aggressive rats and tame rats in both generations. Specifically, it was shown that the expression of the Cacna1b, Drd2, Egr1, and Gad2 genes does not differ between the two generations (88th vs 90th) within each strain, but significantly differs between the strains: in the tame rats of both generations, the expression levels of these genes are significantly lower as compared to those in the aggressive rats. Therefore, these genes hold promise for further studies on behavioral characteristics. Thus, we confirmed polygenic causes of phenotypic manifestation of aggressive reactions.

Stress is an essential part of everyday life. The neuropeptide corticotropin-releasing hormone (CRH, also called CRF and corticoliberin) plays a key role in the integration of neuroendocrine, autonomic and behavioral responses to stress. The activation of the hypothalamic-pituitary-adrenal axis (HPA axis) by neurons of the paraventricular hypothalamic nucleus (PVN), the primary site of synthesis CRH, triggers stress reactions. In addition to the hypothalamus, CRH is widespread in extrahypothalamic brain structures, where it functions as a neuromodulator for coordination and interaction between the humoral and behavioral aspects of a stress response. The axons of neurons expressing CRH are directed to various structures of the brain, where the neuropeptide interacts with specific receptors (CRHR1, CRHR2) and can affect various mediator systems that work together to transmit signals to different brain regions to cause many reactions to stress. Moreover, the effect of stress on brain functions varies from behavioral adaptation to increased survival and increased risk of developing mental disorders. Disturbances of the CRH system regulation are directly related to such disorders: mental pathologies (depression, anxiety, addictions), deviations of neuroendocrinological functions, inflammation, as well as the onset and development of neurodegenerative diseases such as Alzheimer’s disease. In addition, the role of CRH as a regulator of the neurons structure in the areas of the developing and mature brain has been established. To date, studies have been conducted in which CRHR1 is a target for antidepressants, which are, in fact, antagonists of this receptor. In this regard, the study of the participation of the CRH system and its receptors in negative effects on hormone-dependent systems, as well as the possibility of preventing them, is a promising task of modern physiological genetics. In this review, attention will be paid to the role of CRH in the regulation of response to stress, as well as to the involvement of extrahypothalamic CRH in pathophysiology and the correction of mental disorders.

The article presents the results of studying the biodiversity and biotechnological potential of halophilic microorganisms from the thermal highly mineralized Berikey Lake, the salty Lake Tarumovskoye and saline soils of the Peri-Caspian Lowland (Republic of Daghestan). Denitrifying halophilic bacteria of the genus Halomonas and Virgibacillus were identified using microbiological methods and 16S rRNA gene analysis. A new species Halomonas sp. G2 (MW386470) with a similarity of the nucleotide sequences of the 16S rRNA genes is 95 %. Strain G2 is an extreme halophile capable of growing in the range of 5–25 % NaCl (optimum 25 %) and forming a carotenoid pigment. Mesophil, 30–37 °С (optimum 30 °С); neutrophil, pH 6–8 (optimum 7.2–7.4). Strain G2 chemolithotroph; reduces nitrate or nitrite as electron donors; catalase-, amylase-, protease- and β-galactosidase-positive; lipase-, oxidase- and urease-negative. Not able to hydrolyze inositol, indole; produces lysine, gelatin, ectoine; uses citrate and sodium malate as a source of carbon and energy; does not produce ornitin, H₂S or acid from d-mannose, sucrose, glycerol, cellobiose, except for lactose and d-glucose. Susceptible to trimethoprim, ciprofloxacin, ofloxacin, kanamycin, vancomycin, rifampicin, cefuroxime, ampicillin, ceftazidime, fosfomycin, clarithromycin, cefepime, cefaclor. The G+C content in DNA is 67.3 %. A distinctive characteristic of the isolate was the production of industrially significant hydrolytic enzymes such as amylase, protease, β-galactosidase, and oxidoreductase (catalase) at a NaCl concentration of 25 % in the medium. Habitat: saline soils on the territory of the Tersko-Kumskaya lowland (Republic of Daghestan, Russia). The rest of the halophilic isolates of H. ventosae G1 (MW386469), H. elongata G3 (MW386471), V. salinarius B2 (MW386472), and V. salinarius B3 (MW386473) had a high degree of similarity (100 %) with the type strains of H. elongata DSM 2581^Т and V. salarius DSM 18441^Т; the content of G+C in DNA was 65.8, 66.5, 42.8 and 37.3 %, respectively. The strains had a high biotechnological potential at NaCl concentrations of 5 and 25 % in the medium. The data obtained expanded the understanding of the diversity and ecological significance of denitrifying bacteria in the functioning of arid ecosystems and make it possible to identify strains producing enzymes of industrial importance.

72 clinical strains of Klebsiella spp. isolated from samples obtained from humans in Novosibirsk, Russia, were analyzed. Species identification of strains was performed using 16S rRNA and rpoB gene sequences. It was revealed that Klebsiella pneumoniae strains were dominant in the population (57 strains), while the remaining 15 strains were K. grimontii, K. aerogenes, K. oxytoca and K. quasipneumoniae. By molecular serotyping using the wzi gene sequence, K. pneumoniae strains were assigned to twenty-one K-serotypes with a high proportion of virulent K1- and K2-serotypes. It was found that K. pneumoniae strains isolated from the hospitalized patients had a higher resistance to antibiotics compared to the other Klebsiella species. Real-time PCR revealed that the population contained genes of the bla_SHV, bla_TEM, bla_CTX families and the bla_OXA-48 gene, which are the genetic determinants of beta-lactam resistance. It has been shown that the presence of the bla_CTX sequence correlated with the production of extended-spectrum beta-lactamases, and phenotypic resistance to car-bapenems is due to the presence of the bla_OXA-48 gene. At the same time, the carbapenemase genes vim, ndm, kpc, imp were not detected. Among the aminoglycoside resistance genes studied, the aph(6)-Id and aadA genes were found, but their presence did not always coincide with phenotypic resistance. Resistance to fluoroquinolones in the vast majority of strains was accompanied by the presence of the aac(6’)-IB-cr, oqxA, oqxB, qnrB, and qnrS genes in various combinations, while the presence of the oqxA and/or oqxB genes alone did not correlate with resistance to fluoroquinolones. Thus, the detection of bla_CTX and bla_OXA-48 can be used to quickly predict the production of extended-spectrum beta-lactamases and to determine the resistance of Klebsiella to carbapenems. The detection of the aac(6’)-Ib-cr and/or qnrB/qnrS genes can be used to quickly determine resistance to fluoroquinolones.