Vol 13, No 4 (2015)


Assessment of morphometric indexes in the second generation of Scots pine trees in the Chernobyl exclusion zone

Makarenko E.S., Oudalova A.A.


Background. A series of morphometric indexes was studied in Scots pine trees, which are the second generation of trees severely exposed at the Chernobyl accident in doses of 4-5, 10-20 and 80-100 Gy. Materials and methods. Variability of length and mass of needles, curvature of sprouts, tree height and trunk circumference were studied in 2011-2014. Results. Needle gigantism, one of the typical radiomorphoses, was observed in 2012 and 2013 in all groups of trees. Curved sprouts were discovered more often in a reference group. Tree height and trunk circumference in groups of 4-5 and 10-20 Gy were significantly higher than in the reference group. Conclusion. The trees of the second generation of severely exposed pines can be characterized with needle gigantism and stimulation of growth processes (circumference of the trunk, height of the tree) at doses of 4-5 Gy and 10-20 Gy.
Ecological genetics. 2015;13(4):6-8
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Molecular genetic analysis of the spread of resistance to organophosphate insecticides and pyrethroids in populations of the Colorado potato beetle in the Republic of Bashkortostan

Syrtlanova L.A., Kitaev K.A.


Background. We analyzed the spread of mutations in the gene ache - acetylcholinesterase responsible for resistance to the organophosphate insecticides, and in the gene ldvssc1 - electron-sensitive sodium channel, providing kdr-like resistance to pyrethroids in different regions of Republic of Bashkortostan. Materials and methods. Each group included 30-50 overwintered beetles. DNA from beetles was isolated by phenol-chloroform method. Amplification and analysis of genes ache and ldvssc1 were held by bi-PASA (bi-directional PCR amplification of specific allele) according to (Clark et al., 2001). Evaluation of susceptibility to the set of insecticides from classes of POI (aktellik), pyrethroids (decis) was carried out by topical infliction of ethanol solutions of insecticides (1 ml per individual) on the thorax of adults. Diagnostic concentrations of insecticides corresponding with this manner of treatment were determined in the preliminary experiments. Results. In the localities included in the group of southern and eastern regions, frequency distributions coincided. North-western and central group showed excess of phenotypes resistant to organophosphate insecticides compared with a share of genotypes carrying the mutant allele achе, and reducing the incidence of phenotypes resistant to pyrethroids, compared with the frequency of genotypes carrying the mutant allele ldvssc1. Conclusion. The interpretation of these results implies a significant contribution of some unknown component causing the increased level of non-specific resistance to organophosphate insecticides that can be associated with metabolic stability and also with structure and density of the beetle integuments. It should be noted that individuals carrying the mutant allele, providing resistance to pyrethroids, apparently did not have high adaptability, which may be the reason for this discrepancy in the frequencies of occurrence. Polymorphism and mutations in genes controlling metabolism (in particular, a family of genes of cytochrome P 450) should be studied to elucidate the genetic basis of Colorado potato beetle resistance in the territory of the Republic of Bashkortostan.
Ecological genetics. 2015;13(4):9-11
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Cytogenetic status in coal-miners with occupational pulmonary diseases and influence the polymorphisms of XpD and XpG genes

Volobaev V.P., Sinitskiy M.Y., Kulemin Y.E.


Background. Coal-miners are exposed by various harmful factors (chemical agents, ionizing radiation, heavy metals, coal dust). Occupational pulmonary diseases may affect to the level of genotoxic damages. Materials and methods. Venous blood samples extracted from 90 coal-miners with various occupational pulmonary diseases and 26 healthy coal-miners (control 1). Blood samples obtained from 124 non-exposed men were used as the control 2. Assessment of cytogenetic damages was performed usingthe analysis of chromosome aberrations. PCR and gel electrophoresis were used to determine polymorphismsin the genes XpD(rs13181) and XpG(rs17655). Results. We found a significant increase in the frequency of the chromatid- and chromosome-type aberrations in coal-miners and the increase of the chromosomal interchange in miners with occupational pulmonary diseases. The modify effects of polymorphismsin the XpD and XpG genes to the level of chromosome aberrations were discovered. Conclusions. It was shown that the XpD and XpG genes can be used as potential molecular geneticmarkers of increased individual risk of occupational pulmonary diseases.
Ecological genetics. 2015;13(4):12-15
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Pharmacologic correction of paclitaxel-induced geneand myelotoxicity with exstract of Scutellariae baicalensis hairy roots culture

Neupokoeva O.V., Fedorova E.P., Ermolaeva L.A., Filonova M.V., Churin A.A., Voronova O.L., Fomina T.I.


Gene protection effects of Scutellariae baicalensis hairy transgenic roots extract on paclitaxel-induced genotoxic model of mice bone marrow and Drosophila melanogaster somatic cells were exposed in once and course administration during the early and late periods of investigation. Granylocyto- and erythrocytopoiesis stimulation effects of Scutellariae baicalensis hairy roots extract on paclitaxel-induced rats myelosuppression were demonstrated. Materials and methods. Experiments were performed on male and female CBA/CaLac mice and outbred female rats. Paclitaxel (Mitotax®, Dr. Reddy's, India) was administered once in the maximum tolerated dose (MTD): to mice - intraperitoneally, 40 mg/kg, to rats - intravenously, 5 mg/kg. To reduce cytostatic toxicity Scutellariae baicalensis hairy transgenic roots extract was injected per os once at a dose of 200 mg/kg or in 5-day course of 40 mg/kg per day. The state of bone marrow chromosome metaphase plates was examined by the Ford modified method. In the somatic mosaicism test system was used male mutant line, genotype wsn3/Y and female mutant line, genotype yellow of Dr. melanogaster. There were examined 1000 females and marked of number females with single (у, sn3) and double (уsn3) spots. The rat bone marrow was investigated using standard hematological methods. Results. It was shown, that single paclitaxel injection in MTD leaded to generation of genomic and structural disorders in mice bone marrow. In Dr. Melanogaster females, which were grown in nutrient medium with 0,005 % of paclitaxel, appearance of single spots fenotype «y» and «sn» were demonstrated. Paclitaxel injection to rats caused bone marrow hypoplasia in the early periods of investigation. Extract of Scutellaria baicalensis hairy transgenic roots considerably decreased the paclitaxel-induced disorders in mice and rats bone marrow and in Drosophila melanogaster somatic cells.
Ecological genetics. 2015;13(4):16-21
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Sup35 prionization [PSI+] influence the frequency of the gene and chromosome mutations, accounted in the alpha-test in yeast Saccharomyces cerevisiae

Andreychuk Y.V., Zhuk A.S., Inge-Vechtomov S.G., Stepchenkova E.I., Shiriaeva A.A.


Background. A lot of neurodegenerative diseases are coursed by amiloidization of proteins in nerve tissues. In the patients brains suffered from Alzheimer’s disease the high fraction of the nerve cells with abnormal chromosome amount was revealed. There are some data showing that prion form of protein PrP may prevent chromosome segregation in mitosis. But the direct association of prionisation and genome stability was not revealed. Materials and methods. We compared the yeast S. cerevisiae strain bearing the prion form of the termination translation factor Sup35, and the strain with non-prionized Sup35 in the alpha-test system. The model of the alpha-test is based on the mechanism of mating type switching in heterothallic yeast strains. The MAT locus that controls the mating type of yeast cell can be presented by two idiomorphs: the MATalpha and MATa that determine the alpha and a cell types, correspondingly. Only two cells with opposite mating types (alpha × a) could copulate. In the mixture of two yeast strains with alpha mating type the hybrids appears only if one of the parent cells had changed its mating type alpha → a. The mating type switching could course the following genetic events: the loss of the chromosome, gene conversion, recombination, loss of the arm of the chromosome, gene mutations and temporary lesions. These events could be distinguished by using the specially constructed alpha-test system. Results. The [PSI+] strain has showed 2-times decreased frequency of «illegitimate» hybridization in the alpha-test compared to [psi-] strain. But [PSI+] doesn’t influence the frequency of «legitimate» hybridization in the alpha × a crossing. The prion [PSI+] also 2-times reduces the frequency of chromosome loss and gene mutations and increases gene conversion 5-times. This results are also confirmed by the canavanine test. Conclusion. We investigated the effect of the Sup35 prionizaion on the genome stability. Unexpectedly in the [PSI+] strain the frequency of «illegitimate» hybridization was 2-times lower, and frequency of gene mutations and chromosome loss was also reduced. The mechanism of this effect is unclear and requires the further investigation.
Ecological genetics. 2015;13(4):22-24
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Peculiarities of negative consequences of mutagenic action

Nazarenko N.N.


Background. Estimation of correlation between genotype-mutagen interaction and mutagen depression is a main topic of this paper. Materials and methods. Eight winter wheat varieties were treated by gamma-rays and chemical mutagens. The frequency and spectrum of chromosomal aberrations, parameters of field growing and development were investigated. Results. Frequency of chromosomal aberrations, plant survival and some parameters of yield structure are reliable indicators of mutagen depression. Repeat action of same mutagen lead to decreasing of chromosomal aberrations number. Conclusion. Mutation varieties are less sensitive to some types of mutagen action.
Ecological genetics. 2015;13(4):25-26
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Genotoxic effect of fipronil on somatic and germ cells of mice

Lovinskaya A.V.


Background. It is known that many pesticides can act as mutagens by causing cytotoxic and negative genetic effects. Fipronil is insecticide, which widely used marketed under the trade names Adonis and Regent for locust control in Kazakhstan. Materials and methods. It studied mouse cells of internal organs (liver, lungs, spleen, testes) using the Comet assay and synaptonemal complex analysis in spermatocytes using immunocytochemistry methods. Results. It has been found that fipronil damages DNA in cells of the lung, spleen and liver intoxicated animals. Fipronil has a negative effect on the germ cells of mice, causing damage to synaptonemal complex structure of spermatocytes. Conclusion. The studies have shown that fipronil has genotoxotic effect.
Ecological genetics. 2015;13(4):27-29
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The assessment of micronucleus frequency in lymphocytes in the cohort of coal-miners characterized by different polymorphisms of double strand break reparation genes

Sinitsky M.Y., Volobaev V.P., Asanov M.A.


Background: Coal-miners are exposed to a lot of number of harmful factors (chemical agents, ionizing radiation, heavy metals, coal dust etc.). Material and methods: Venous blood samples extracted from 129 coal-miners. Assessment of cytogenetic damage was performed using the cytokinesis-block micronucleus assay (CBMN) on peripheral blood lymphocytes. PCR and gel electrophoresis were used to determine polymorphisms in the genes Lig4 (rs1805388) and XRCC4 (rs6869366). Results: We found a significant increase in the frequency of binucleated lymphocytes with micronuclei (MN) and protrusions in carriers of the Ile/ Ile genotype of the Lig4 gene Thr9Ile polymorphism in comparison to Thr/Thr and Thr/Ile genotypes. Conclusions: Thr9Ile polymorphism within Lig4 gene can be used as potential molecular genetic markers of increased individual susceptibility to the complex of harmful factors in coal-mining conditions.
Ecological genetics. 2015;13(4):30-33
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Markers of human extraembryonal cells individual radiosensitivity in vitro

Belenko A.A., Vasilyev S.A., Lebedev I.N.


Background: Genotoxic effects of ionizing radiation in early stages of human embryonic development can lead to fatal consequences. At the same time, the radiosensitivity of human embryonic and extraembryonic cells is still poorly studied. In this study, the analysis of DNA double-strand break repair effectiveness in human extraembryonal fibroblasts was carried out. Materials and methods. Extraembryonic human fibroblasts was irradiated by 1 Gy gamma-rays using Theratron Equinox (Cancer Research Institute, Tomsk). The level of DNA double strand breaks was assessed using γH2AX foci. Frequency of cytogenetic damage was assessed using micronucleus test conducted with FISH as a frequency of centromere-negative micronuclei. Results. No significant correlation was observed between both endogenous and residual levels of radiation-induced γH2AX foci and frequency of micronuclei after irradiation. It is suggested to be a result of the specificity of extraembryonal fibroblast radiation-induced response. The spontaneous frequency of centromere-negative micronuclei correlated with radiation-induced frequency of centromere-negative micronuclei. Conclusion. It was shown that human extraembryonal fibroblasts ability to repair radiation-induced DNA damage is likely to be reflected by the repair of spontaneous DNA double strand breaks.
Ecological genetics. 2015;13(4):34-36
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The genetics of the traits determining adaptability to abiotic stress in rice (Oryza sativa L.)

Kharitonov E.M., Goncharova Y.C., Maluchenko E.A.


Most of rice cultivationarea in Russia, characterized by unfavorable soil conditions: salinity varying degrees and types, alkalinization, lack of mineral nutrients. In addition, in the Kuban in one of the most Northern region of rice cultivation during of rice the growing season marked as high more than 40 °C and low temperature 0 °C. Consequently, to increase the production of culture less necessary increasing potential productivity then stability of yield, and complex of resistance to stress genes. The article presents data on polymorphism and the molecular marking of characteristics defining adaptability to various abiotic stresses.
Ecological genetics. 2015;13(4):37-54
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Genetic diversity comparative evaluation of Pinus Sylvestris L. and Picea x Fennica (regel) kom. native populations and clonal seed orchards in russian Karelia

Ilinov A.A., Raevsky B.V.


Genetic diversity levels in 4 native populations of Finnish spruce and Scots pine each and 2 fields of conifer seed orchard growing in Karelia have been investigated using microsatellite loci. As a result high levels of basic genetic diversity parameters have been revealed for native populations of both species. It was found that expected heterozygosity figers calculated for the populations investigated were higher than the observed ones. This case thereby indicates a deficit of heterozygotes in the Karelian pine and spruce populations. Genetic diversity figures found for spruce seed orchard were much lower than for native populations of Picea x fennica. This fact, in our opinion, reflects the unsufficent representation of genetic pool both within the seed orchard field investigated and in spruce plus trees' breeding population on the whole. Scots pine seed orchard has been characterised by a high level of genetic diversity matched to native populations one.
Ecological genetics. 2015;13(4):55-67
pages 55-67 views

Mitochondrial DNA d-loop polymorphism analysis for estimation of diversity in chicken flocks of Pavlov breed

Demin A.G., Danilova M.I., Galkina S.A.


Elucidation of the complex origin of various chicken breeds and populations is of essential importance for understanding, preserving and exploiting their genetic diversity. Here, we aim to assess different contributions to mitochondrial genetic diversity of Pavlov chicken breed. Mitochondrial DNA control region (D-loop of 1231/1232 b. p. length) in 37 chickens of Pavlov breed was sequenced. Individuals were selected from three flocks belonging to Federal State Unitary research farm “Gene Pool” (Genofond), Pushkin, Leningrad region, to the collection farm of All-Russian R & D and Technology Institute of Poultry Industry (GNU VNITIP), Sergiev Posad, Moscow region, and to fancy breeders from Barnaul (Altai region). The Pavlov chicken D-loop sequences were compared with D-loop sequences annotated in GenBank for established chicken haplogroups. We have found eleven haplotypes belonging to two haplogroups (E1 and A). Genetic uniformity and stability have been shown for the GNU VNITIP and Barnaul flocks, while D-loop high polymorphism was found in the population from the research farm “Gene Pool”. There appears a tendency for genetic fragmentation of Pavlov chicken breed.
Ecological genetics. 2015;13(4):68-75
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Molecular phylogenetic analysis of the tonoplast H+-ATPase subunits

Chen T., Mikhaylova Y.V., Shishova M.F.


Vacuolar H+-ATPase is a multi-subunit protein complex, which fulfills a number of crucial functions in plant cell. Different mechanisms are known to be important for the regulation of proton-transporting enzyme activity at transcriptional and post-translational levels. In this investigation we performed a comparison analysis of molecular phylogeny of different subunits of vacuolar H+-ATPase directed in the elucidation of conservative rate of membrane and peripheral complexes of the enzyme. High rate of conservatism was shown for subunits c, d and В, as well as ancient duplications of subunit a.
Ecological genetics. 2015;13(4):76-90
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The role of XRCC1, XRCC3 and PALB2 genes in the genesis of breast cancer

Kipen V.N., Melnov S.B., Smolyakova R.M.


Background: this study describes the contribution of XRCC1, XRCC3 and PALB2 genes in the genesis of sporadic forms of breast cancer in Belarus patients. Materials and Methods: this study included 169 patients with sporadic breast cancer and 185 healthy patients. The molecular genetic analysis was performed by RFLP and PAGE electrophoresis. Results: in this study we identified genotypes of high risk breast cancer patient’s for PALB2, XRCC1 and XRCC3 genes. We also discovered a statistically significant associations between the GG genotype for p.Q399R (XRCC1) and tumor grade, and between TT genotype for XRCC3 gene and the presence of metastases in the regional lymph nodes. The patients with TT genotype (p.T241M, XRCC3) showed the fivefold increase in the risk of local breast cancer recurrence. Conclusion: For the first time it was determined the prevalence of polymorphic variants of the DNA repair genes: p.Q399R (XRCC1), p.T241M (XRCC3), p.T1100T (PALB2) and p.Q559P (PALB2) for Belarus patients with the breast cancer. Analysis of SNPs in the DNA repair key genes may have clinical and prognostic significance during the formation of high risk patient’s groups developing the breast cancer.
Ecological genetics. 2015;13(4):91-98
pages 91-98 views

High hydrostatic pressure influence on viability and mutagenesis of Salmonella Typhimurium

Karamova N.S., Zelenikhin P.V., Kiselev V.D., Lipatnikova A.A., Ilinskaya O.N.


Background: pressure is a well-known physical environmental parameter. Nevertheless, the basic principles of microbial survival under high hydrostatic pressure (HHP), especially genetic response to pressure, are still poorly understood. The purpose of this study was to investigate the influence of HHP ranging from 50 to 800 MPa on viability and mutagenesis of Salmonella typhimurium. Materials and methods. The standard plate count method (counting the total number of colony forming units (CFUs) on the plate) and the propidium iodide (PI) flow cytometric assay were used to determine the bacterial viability after HHP treatment. Ability of HHP to induce gene mutations was examined by the Ames assay employing Salmonella typhimurium TA100 and TA98. Results. The results obtained showed that survival of S. typhimurium cells considerably decreased when bacteria were exposed to a pressure of 200 MPa and above. Herewith, the survival index calculated according to the total number of CFUs was up to six orders of magnitude lower than that obtained by the flow cytometric analysis under the same HHP. This fact can be explained by the entrance of the some part of bacterial population into the viable but nonculturable (VBNC) state. The pressure of 50 MPa was found to cause a 1.9-fold increase in the number of His+ revertants of S. typhimurium TA98 in Ames test. Conclusion. Our results demonstrate that HPP of 200 MPa and above significantly inhibits the viability of S. typhimurium cells as well as triggers the induction of VBNC state. The results of Ames test suggest that HHP of 50 MPa can induce gene mutations in bacterial cells. The possible mechanisms of HHP effects on cells viability as well as genetic response of bacteria under HHP are discussed.
Ecological genetics. 2015;13(4):99-107
pages 99-107 views

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