Vol 15, No 1 (2017)

Predator odor induces genome instability in the mouse bone marrow cells
Glinin T.S., Starshova P.A., Shubina V.A., Anisimova M.V., Bondarenko A.A., Moshkin M.P., Daev E.V.

Background. Long coevolution of prey and predator species of mammals creates specific mechanisms of their interaction, e. g. prey’s innate behavior aversive to the predator odor. However, little is known about genetic responses in the prey organism. We assessed genome instability of the bone marrow cells in mice affected by the cat’s odor influence, and proposed pathway of such action.

Materials and methods. CBA mouse males were exposed to volatiles from adult cat urine for 2 or 24 hours. To estimate the genetic effect, ana-telophase method of chromosome aberration analysis and comet assay were used. The level of corticosterone was also measured after the exposure for 30 or 60 minutes.

Results. The exposure to cat’s urine volatiles for 2 hours induced damage of DNA in bone marrow cells of the mouse males as was shown by the DNA comet analysis. The exposure for 24 hours elevated the frequency of chromosome aberrations in mitotically dividing cells at ana-telophase stage. No significant changes were found in the level of corticosterone in the peripheral blood.

Conclusion. We have shown that volatile chemosignals from predator’s urine induce genomic instability in bone marrow cells of a prey. The hormonal pathway of such influence is still unknown. Intraorganismic paths leading to genome damage are discussed as well as far consequences of discovered effects.

Ecological genetics. 2017;15(1):4-11
Productivity and stress-tolerance of transgenic tobacco plants with constitutive expression of rapeseed glutathione synthetase gene BnGSH
Kuluev B.R., Berezhneva Z.A., Mikhaylova E.V., Postrigan B.N., Knyazev A.V.

Summary: Glutathione is the most important part of plant antioxidant defense system. Biosynthesis of glutathione in the cells is performed by two enzymes: glutamylcysteine ligase and glutathione synthetase, the latter catalyzing the attachment of glycine to a dipeptide glutamylcysteine. In literature there is information on the improvement of heavy metal-tolerance of transgenic plants due to the increase in the expression level of glutathione synthetase genes. However there is not enough data on the tolerance of these plants to other types of abiotic stress. Therefore the aim of our research was to make transgenic tobacco plants with constitutive expression of rapeseed glutathione synthetase gene BnGSH and to estimate their growth parameters in normal conditions and under salt, drough and cold stress. Using agrobacterial transformation method, we generated 17 lines of transgenic plants containing rapeseed BnGSH gene under control of 35S promoter. The presence of transgenes was confirmed by PCR method and histochemical analysis of the activity of GUS reporter gene. 12 lines with the highest expression of BnGSH gene were chosen on the basis of the results of RT-PCR. We performed morphological analysis, including measurements of stem hight, leaf area, flower length, fresh and dry weight of shoots and root length. Some transgenic plants demonstrated increased productivity in normal conditions as well as under NaCl stress. However, no change in drought and cold tolerance was observed in transgenic plants.

Ecological genetics. 2017;15(1):12-19
Diversity of the species of genus Avena revealed by morphological characters and resistance to Fusarium infection of grain
Gagkaeva T.Y., Gavrilova O.P., Orina A.A., Blinova E.V., Loskutov I.G.

Sixty six genotypes belong to wild and cultivated Avena species from the VIR collection were evaluated for infection of grain by Fusarium fungi and mycotoxins in accumulation. Among genotypes 13.6, 28.8, and 57.6% were diploid, tetraploid, and hexaploid oats, respectively.

The aim of this study was to recognize the interrelationship between the wild species of the genus Avena not subjected to formal breeding programs, and Fusarium fungi, which have been reported as predominant seed borne pathogens. The real-time PCR was used to simultaneously detect and quantify fungi in grain of Avena genotypes. Mycotoxin analysis was performed by ELISA. The average amount of Fusarium DNA and deoxynivalenol (DON) in groups of the tetraploid oats were higher than they were found in the groups of di- and hexaploid species. It was determining the strong correlation between the presence of Tri-Fusarium DNA and DON content (p < 0,01). The low amounts of DON in the grain were detected in seven hexaploid genotypes (A. byzantina, A. sterilis, A. sativa, and A. fatua) and one diploid A. wiestii. The connection of morphological characters (weight of 1000 grains, husk hardness, trichomes profusion and plant height) of Avena species and the indicators of Fusarium infection were analyzed. Only biomass of Fusarium has demonstrated significant connection with weight of 1000 grains. Significant correlation between the hardness of husk and the amounts of Fusarium DNA (p < 0,01) was detected. For Fusarium the strong protective effect of the husk from the penetration of pathogens into grain was evident.

Ecological genetics. 2017;15(1):20-29
Low level of genetic differentiation among populations of the rare species Allium regelianum A.K. Becker ex Iljin from the Volgograd region detected by ISSR-analysis
Trifonova A.A., Kochieva E.Z., Kudryavtsev A.M.

Background. Knowledge of genetic diversity within and among populations of rare and endangered plants species is practically important for conservation management. Molecular markers are useful tools for analysis of genetic diversity. In this study ISSR-analysis of rare endemic species Allium regelianum which grows in the Volgograd region was performed for the first time.

Materials and Methods. A total of 93 samples from the 11 populations were collected and used in analysis. Six primers used in ISSR-analysis. Data analysis was performed using the GenAlEx 6.41, POPGEN 1.32, PAST 3.11 and STRUCTURE 2.3.1 programs.

Results. A total of 109 ISSR-fragments were scored of which 87 (79.8%) were polymorphic. Comparatively high level of intrapopulation diversity was estimated for the population of the area near Krasnoyarskii khutor, Serafimovichskii administrative region, and for the population of Khrenovatyi Liman, Nikolaevskii administrative region of Volgograd Oblast. Genetic similarity index among populations ranged from 0.88 to 0.96. Genetic differentiation among populations of A. regelianum, GST was 0.284, only. Analysis of molecular variance showed that genetic heterogeneity of A. regelianum 83% was attributed to differences within populations and 17% occurred among populations. Principal coordinate analysis and analysis of populations structure (with the used of STRUCTURE program) found no clear differentiation among populations.

Conclusion. The estimation of intra- and interpopulation diversity of A. regelianum was performed. ISSRs detected high levels of genetic similarity within the populations of A. regelianum and low level of genetic differentiation among populations.

Ecological genetics. 2017;15(1):30-37
Genetic polymorphism of natural isolates of Nosema Pyrausta (Microsporidia: Nosematidae)
Grushevaya I.V., Ignatieva A.N., Malysh J.M., Trepashko L.I., Tokarev Y.S., Frolov A.N.

Background. Microsporidia are ubiquitous parasites of animals, most abundant in arthropods and fishes. Many species of these parasites are important from standpoints of medicine, veterinary and agriculture. Microsporidium Nosema pyrausta is an important disease agent in corn borer populations causing adverse effect on host fitness. Genotyping of this parasite is necessary for proper species identification and intraspecific polymorphysm studies.

Materials and Methods. Microsporidia-infected larvae of corn borers of the genus Ostrinia were recovered from Krasnodar Territory in Russia and Gomel Region in Belarus. Small subunit ribosomal RNA (SSU rRNA) was amplified and sequenced directly, while intergenic spacer (IGS) was amplified, cloned and sequenced (1-5 clones per sample) for four isolates of microsporidia. Sequences were aligned and compared using standard bioinformatics tools (Clustal W and BLAST).

Results. SSU rRNA genotyping showed allocation of all four isolates to N. pyrausta with 100% identity to each other and 99.7% similarity to Nosema bombycis, the type species of the genus Nosema. High levels of IGS sequence variation (61-74%) is observed both between isolates of different species and populations of microsporidia as well as between molecular clones within parasite isolates from individual hosts.

Conclusion. N. pyrausta is widespread in corn borer populations and its genetic structure is complicated, as in other species of these parasites. Further studies of molecular markers are needed for genetic differentiation of geographic isolates of N. pyrausta.

Ecological genetics. 2017;15(1):38-43
Prevalence rates of Wolbachia endosymbiotic bacterium in natural populations of Ostrinia Nubilalis and Ostrinia Scapulalis (Lepidoptera: Pyraloidea: Crambidae) in South-Western Russia
Tokarev Y.S., Yudina M.A., Malysh J.M., Bykov R.A., Frolov A.N., Grushevaya I.V., Ilinsky Y.Y.

Background. Endosymbiotic bacteria of the genus Wolbachia are widespread in arthropods and often cause reproductive abnormalities in lepidopteran insects, including corn borers of the genus Ostrinia. Wolbachia-Ostrinia is a promising model for studies of parasite-host interactions yet parasite prevalence in natural insect host populations remains unknown.

Materials and Methods. Molecular genetic screening and statistical analysis is applied to evaluate prevalence rates of Wolbachia in sympatric populations of two corn borer species. Individual genomic DNA samples were extracted from last instar larvae collected in nature from different forage plants. For each sample of DNA showing positive signal with insect-specific primers the detection is performed using three diagnostic loci of Wolbachia: 16SrRNA, gatB and fbpA.

Results. Wolbachia-positive signal is obtained for 13.5% larvae of Ostrinia nubilalis (N = 141) and 31.9% larvae of Ostrinia scapulalis (N = 138). In different localities the Wolbachia prevalence ranged from 2.9% (N = 34) to 65.8% (N = 38). Significantly higher rates of Wolbachia prevalence in insects from mugwort and hemp (O. scapulalis) as compared to those from corn (O. nubilalis) are revealed in three out of four localities.

Conclusions. Endosymbiotic bacteria of the genus Wolbachia are revealed in natural populations of corn borers for the first time for Eastern Europe. The prevalence rates can be high and this should be taken into consideration when reproductive isolation is examined in population of these hosts as well as establishment of laboratory cultures is performed.

Ecological genetics. 2017;15(1):44-49
R577X polymorphism of alpha-actinin-3 in human populations of North-Eastern Asia
Malyarchuk B.A., Derenko M.V., Denisova G.A.

Background. In polymorphism rs1815739, a C → T transition converts arginine to a premature stop-codon at residue 577 of the alpha-actinin-3 (ACTN3) protein (R577X polymorphism). This polymorphism may affect muscle performance, and the derived 577X allele has been found to be under-represented in sprint/power athletes. In addition, loss of alpha-actinin-3 results in a shift in muscle metabolism toward the more efficient aerobic pathway, thus pointing that this polymorphism may have been involved in enhancing the capability for hunting and for cold adaptation. Here, we study rs1815739 polymorphism in native populations (Chukchi, Koryaks and Evens) and newcomers (Russians) of North-Eastern Asia.

Materials and methods. Genomic DNA was isolated from peripheral blood. ACTN3 genotypes for rs1815739 locus were established by enzymatic digestion of amplicons with DdeI. Heterozygotes TT were confirmed by DNA sequencing. In addition, data on exome variation in Siberian populations were analyzed.

Results. Lowered frequencies (less than 40%) of “mutant” allele rs1815739-T were found in studied populations of North-Eastern Asia. Analysis of exome data has shown that haplotype comprising the rs1815739-T allele reaches the highest frequencies in populations of Southern and Central Siberia, while it is rather rare in the north-east of Siberia.

Conclusion. The results obtained contradict the hypothesis that the rs1815739 polymorphism may have been involved in cold adaptation of North-East Siberians.

Ecological genetics. 2017;15(1):50-56
Investigation of paraoxonase 1 activity of the workers at the plant, who have long-term contact with organophosphorus compounds
Razgildina N.D., Miroshnikova V.V., Fomichev A.V., Malisheva E.V., Panteleeva A.A., Pchelina S.N.

Background. Liver enzyme paraoxonase 1 (PON1) plays an important role in protection the organism from toxic effects of organophosphorus compounds (OPs) via their hydrolysis whose rate and efficiency depend on PON1 serum level activity. PON1 activity is largely determined by the polymorphic variants of the PON1 gene. Effect of long-term work with exposure to the toxic OPs on the PON1 activity is almost unknown. The aim of the present work was to study the effect of long-term work with exposure to the toxic OPs on PON1 serum enzymatic activity depending on polymorphisms Q191R, L54M, C(-108)T PON1 gene.

Materials and methods. PON1 serum enzymatic activity and PON1 polymorphisms were determined in men, who were categorized in 2 groups: workers of companies providing storage and disposal of the OPs (68) and control group (37). The PON1 191, PON1 55 and PON1 108 polymorphisms were studied by polymerase chain reaction/restriction fragment length polymorphism. PON1 serum enzymatic activity was measured by a spectrophotometric method using paraoxon.

Results. PON1 activity in workers with exposure to the toxic OPs relative was increased compared to the control group (p = 0,027). Differences in serum PON1 activity was shown for the carriers of certain genotypes of the PON1 gene: PON1 serum activity was higher in workers compared to controls only for LL genotype (L54M polymorphism) and C allele (C(-108)T polymorphism) carriers (p < 0,001 and p = 0,002, correspondently).

Conclusion. We suggest that the increase in serum PON1 activity in workers providing storage and disposal of OPs could be modulated with the polymorphic variants of the PON1 gene.

Ecological genetics. 2017;15(1):57-63
Retsenziya na knigu S.K. Abileva i V.M. Glazera «Mutagenez s osnovami gentoksikologii»
Inge-Vechtomov S.G.


Ecological genetics. 2017;15(1):64-64

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