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Vol 19, No 1 (2021)

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Genetic basis of ecosystems evolution
The expansin gene NtEXPA5 increases stress tolerance of tobacco hairy roots through an effect on the antioxidant system
Kuluev B.R., Musin K.G., Yakupova A.B.

Background: Expansins are non-enzymatic proteins involved in the softening of cell walls, the mechanism of action of which is associated with the weakening and breaking of hydrogen bonds between xyloglucans and cellulose microfibrils and is aimed at ensuring cell expansion.

The aim of our work was to obtain hairy roots of tobacco with constitutive expression of the NtEXPA5 expansin gene, their morphometric analysis and assessment of the state of their antioxidant system in response to stress factors.

Materials and methods: The hairy roots were obtained from transgenic tobacco plants expressing the NtEXPA5 gene under the control of the 35S promoter.

Results: Constitutive expression of the NtEXPA5 gene promoted an increase in the length and dry weight of hairy roots both under normal conditions and under the action of salinity, copper sulfate, cadmium acetate, and mannitol. Both under normal conditions and under the action of stress factors in transgenic hairy roots, an increase in the activity of superoxide dismutase and the total antioxidate activity was recorded.

Conclusion: Expansins exert their positive effect on the productivity and stress tolerance of plants not only through their influence on cell expansion, but also through the effect on the antioxidant system.

Ecological genetics. 2021;19(1):5-12
Creation of an inducible vector system based on the rhizobia nodA gene promoter
Chubukova O.V., Vershinina Z.R., Matnyazov R.T., Baymiev A.K., Baymiev A.K.

Background: The possibility of changing the properties of rhizobial bacteria by giving them the ability to regulate the expression of additionally introduced genes into them is an urgent task both for fundamental science and for applied agrobiology, since this will make it possible to obtain microsymbionts with desired properties. An expression construct using the rhizobia regulatory system was created in this work. The rhizobia nodD gene encodes a regulatory protein that, in the presence of plant inducers, flavonoids, activates the transcription of nod-genes involved in the early stages of the formation of legume-rhizobium symbiosis.

Materials and methods: A vector construct containing the nodD gene from Rhizobium leguminosarum bv. trifoli under the regulation of its own promoter and the gfp gene under the regulation of the nodA gene promoter from the same rhizobia was obtained. Neorhizobium galegae CIAM 0702 were transformed with the vector construct.

Results: It has been shown that in recombinant strains synthetic flavonoids are capable of inducing expression of gfp gene to varying degrees.

Conclusion: In the future, the results can be used to obtain rhizosphere microorganisms with a controlled synthesis of growth-stimulating and protective substances.

Ecological genetics. 2021;19(1):13-21
Genetic diversity of Scots pine trees of different selection categories in plus stands of Karelia
Ilinov A.A., Raevsky B.V.

Background: Genetic control is necessary at all stages of forest selection for the effective use of objects of unified genetic-selection complex (UGSC). This is particularly important for performing a breeding inventory. The aim of the present study was to evaluate the genetic variability of trees of different selection categories in several plus stands of Scots pine.

Materials and methods: 116 trees of Scots pine of various selection categories have been selected from four plus stands within the middle taiga subzone of Karelia. The analysis of the pine samples had been performed using four nuclear SSR loci tests. DNA fragments were separated on a sequencer CEQ 8000. The main criteria of the genetic diversity and F-statistics were calculated. The genetic structure of the selection groups was evaluated using the coefficient of genetic originality (CGO).

Results: All the selection groups evaluated with CGO were characterized by a low content of alleles, which are rare for this part of the Scots pine areal. The plus trees were characterized by a reduced level of genetic diversity (Ho = 0.48–0.59; He = 0.47–0.59) as compared to minus (Ho = 0.46–0.64; He = 0.49–0.65) and normal trees (Ho = 0.50–0.69; He = 0.49–0.65). However, the observed differences between the breeding groups were not statistically significant.

Conclusion: The absence of significant differences between the selection groups indicates the need of a separate assessment of the genetic and economic value of plus trees according to their phenotype. The obtained data is necessary for the organization of objects of UGSC, testing of clonal and seed offspring of plus trees, certification of clones, etc.

Ecological genetics. 2021;19(1):23-35
Searching for regulators that interact with BELL1 transcription factor and control the legume-rhizobial symbiosis development
Dolgikh A.V., Dolgikh E.A.

The development of nitrogen-fixing nodule, which is formed during legume-rhizobial symbiosis, requires the involvement of cell cycle regulators, phytohormones and homeodomain-containing transcription factors as well as other organogenesis processes. Along with homedomain-containing transcription factors from KNOX family, which participation in the control of nodule development has been studied recently, the role of transcription factors from BELL family in this process remains under-explored. At the same time, transcriptomic data for legumes shows an increase in the expression levels of genes encoding proteins of this family in the plant roots inoculated by Rhizobium. In this work we performed screening of сDNA library, which was generated from inoculated pea roots, using BELL1 protein in yeast two-hybrid system. As a result, we detected two interacting proteins, which are highly interesting for future examination. In the first case we discovered interaction between BELL1 and LysM-containing receptor-like kinase LYK9. Another identified regulator interacting with BELL1 became the glycine-rich protein A3, which may be involved in the regulation of defense reactions in plants and their resistance to phytopathogens. Transcriptomic analysis for pea roots has revealed high expression level of gene, which encodes this protein in the nodules, that may demonstrate its important role in symbiosis regulation.

Ecological genetics. 2021;19(1):37-45
Genetic toxicology
Thermophilic aerobic organoheterotrophic soil bacteria from anthropogenically changed territories of Saint Petersburg and Leningrad region
Zhuravleva A.S., Volkova E.N., Galushko A.S.

Anthropogenically altered soils of Saint Petersburg and Luga (Leningrad Region) were investigated for the presence of thermophilic aerobic chemoorganoheterotrophic bacteria, potentially capable of decomposing hydrocarbons at elevated temperatures (60 °C). 6 strains of pure spore-forming cultures of bacteria were isolated. Analysis of the nucleotide sequences of the 16S rRNA genes showed that they belong to the genera Geobacillus and Aeribacillus. For the first time, we obtained information on the presence of representatives of the genus Aeribacillus, which are typical inhabitants of hot springs and zones with geothermal activity, in the soils of the regions of Saint Petersburg and the Leningrad Region.

Ecological genetics. 2021;19(1):47-58
Human ecological genetics
The frequency of chromosomal aberrations in persons chronically in contact with the factor of industrial exposure of the aluminum plant
Volobaev V.P., Lytkina E.S., Schetnikova E.A., Vdovina E.D., Mikheeva E.E., Gurevich E.B., Mishchenko E.P., Larionov A.V.

Background: Chronic excess human exposure to fluorides is an underestimated problem. There is information that fluorides are capable of exerting a genotoxic effect in vitro and in vivo. The data obtained on human populations are limited and contradictory. More research is needed.

Materials and methods: The material for the study was peripheral venous blood taken from residents of the city of Novokuznetsk. The material was selected in four groups: workers of the Novokuznetsk Aluminum Plant (NkAZ) with fluorosis, NkAZ workers without pathology, residents of the area adjacent to NkAZ, residents of the city living away from the industrial zone. The frequency of chromosomal abnormalities in blood leukocytes was estimated using the method of accounting for metaphase chromosomal aberrations, as well as the concentration of fluoride ion in blood plasma by direct potentiometry.

Results: A significant increase in the frequency of aberrant metaphases in blood leukocytes was observed among workers at an aluminum smelter compared to control, workers with fluorosis compared to workers without pathology, and residents of the area adjacent to the plant compared to residents of the city living far from the industrial zone. A correlation of the frequency of chromosomal aberrations with the concentration of fluoride ion in blood plasma was noted.

Conclusion: The results obtained confirm the ability of the industrial factor of the aluminum smelter to exert a genotoxic load on the human population. The presence of an association of fluoride factor with chromosomal abnormalities in blood leukocytes is likely.

Ecological genetics. 2021;19(1):59-66
Prognostic capacity of the HIrisPlex genetic phenotyping system in the Belarusian population
Seredenko M.V., Vakula S.I., Shapturenko M.N., Kondratsiuk A.V., Borovko S.R., Lugovnjov A.V., Gudievskaya I.G., Skrypnik O.V., Marchenko L.N., Kilchevsky A.V.

Background. Externally visible traits such as iris and hair color are of interest in variants of arias of research from archeology and population genetics to forensic science. Recently a number of models have been developed to predict variants of human phenotypic characteristics. The most popular of them is the HIrisPlex system consisting of 24 SNPs, associated with human eyes and hair color.

Materials and methods. We used the HIrisPlex system for phenotyping of the Belarusian population. Allelic variants of the SNPs were genotyped by massive parallel sequencing. The analysis of the obtained data was performed using the free-available online resource

Results. The comparison of the results of genotyping with actual data showed high accuracy of the system in determining blue and brown eyes (94.5% and 91.8% of correct predictions, respectively), and in identifying brown-haired (84.7%) and red-haired individuals (80%). The HIrisPlex system successfully  predicts iris and hair color in two most numerous phenotypic groups - "brown-haired with blue eyes" (BA=78.5%) and "brown-haired with brown eyes" (BA=85.5%).

Conclusion. The HIrisPlex system has a high prognostic ability  for some phenotypic characteristics in the Belarusian population. But for traits that are difficult identified, additional researchers are required.

Ecological genetics. 2021;19(1):67-76
Studying the genotoxic effects of the working environment on the workers of a coal-fired power plant using a micronuclear test on blood lymphocytes
Fedoseev V.I., Stepanov D.D., Minina V.I.

Background: The micronucleus test of peripheral blood lymphocytes provides a qualitative assessment of the state of the genome. A study was carried out aimed at studying the influence of factors of the working environment of coal-fired power plants on the state of the DNA of working data of enterprises.

Materials and methods: Using a micronucleus test with a cytochalasin block on human blood lymphocytes in vitro, 116 men were examined: 45 employees of the Novo-Kemerovo coal-fired power plant and 71 healthy residents of the Kemerovo region.

Results: An increase in the frequency of occurrence of cells with micronuclei, bridges and protrusions in the blood lymphocytes of workers of a coal-fired power plant in comparison with healthy residents of the same area was found.

Conclusion: The data confirm the existing idea of the potential danger of combustion products for human health.

Ecological genetics. 2021;19(1):77-88
Opinions, discussions
Regulatory status of genome-editing plants: perspectives for Russian Federation
Bogatyreva N.V., Sokolov A.Y., Moiseeva Y.M., Gusev Y.S., Chumakov M.I.

The high-quality regulatory support for the use of plant genome editing technology is an urgent scientific and practical task of modern agriculture. Currently, the status of plants obtained using genomic editing (GE) technologies is not defined in Russian legislation. The article describes the principles and mechanism of CRISPR/Cas9 technology, and discusses the biological safety of the GE-plants. Fundamentally different approaches to genetically modified (GM) and GE-plants in the world are analyzed. We discuss the problems and contradictions of extending the GM-plants legal regulation to GE-plants. In particular, the European Court of Justice decision that extended the European GM-plants legislation for GE-plants. It is proposed to determine the legal status of GE-plants in Russian legislation, taking into account existing international practices, and protect the interests of the government in the field of biological and food security.

Ecological genetics. 2021;19(1):89-101

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