Vol 15, No 2 (2017)

Study of allele pool and genetic structure of russian population of lowland-caucasian line of European bison (Bison bonasus)
Dotsev A.V., Aksenova P.V., Volkova V.V., Kharzinova V.R., Kostyunina O.V., Mnatsekanov R.A., Zinovieva N.A.

Summary: Background. The European bison (Bison bonasus) is the only wild ox of Europe, survived to our days. Whilst numerous stu dies have been undertaken to characterize the Lowland line of European bison, it is little known about allele pool and population genetic structure of the Lowland-Caucasian line of wisent.

Materials and methods. The samples were collected from twenty-six animals of Russian breeding nucleus of Lowland-Caucasian line. Ten Bos Taurus microsatellites (TGLA227, BM2113, ETH10, SPS115, TGLA122, INRA23, TGLA126, BM1818, ETH225, and BM1824) were used for analysis.

Results. Eight of ten microsatellite loci (excluding TGLA227 and INRA23) were polymorphic. The number of alleles per locus is varied of one to five with average value of 2.80 ± 0.47. The alleles, which are specific for Lowland-Caucasian line, were identified. We observed relatively high inbreeding level (FIS = 0,091) and very low effective population size (Ne = 1.8, 95% CIs, Parametric 1.1-2.9). We showed that two genetically distinct groups have taken part in formation of allele pool of studied wisent population.

Conclusion. Our data indicated that the development of breeding program to decrease the inbreeding degree and to increase the level of genetic diversity is necessary.

Ecological genetics. 2017;15(2):4-10
The role of environmental factors in the formation of the genetic structure of P. аbies populations
Zakharova K.V., Seits K.S.

Summary: Background. Norway spruce (Picea abies (L.) Karst.) is one of the main forest forming spruce species in Europe. Their populations are characterized by the genetic heterogeneity between individuals in population. The genetic structure of P. abies populations studied supported a hypothesis of an environment impact on the degree of genetic differentiation among populations.

Materials and methods. Analysis of genetic diversity of natural populations in distinct geographical regions from North-West Russia, South-West Russia, South Norway were done using nuclear microsatellites. The needles were collected from the 20-30 adult trees in each geographical spot taking into account the local environment. The statistical calculations were performed with GenAlEx 6.5.03 and Structure 2.3.4.

Results. The analysis highlighted the environmental impact on the genetic diversity. The genetic structure of spruce cenopopulations from poor conditions are extremely different from those of cenopopulations growing under rich environmental conditions in one geographic region. The genetic differences between them are comparable with the genetic differences between spatially remote populations.

Conclusion. The role of the isolation by environment as a pattern in which genetic differentiation increases with environmental differences independent of the geographic distance is essential for genetic structure of spruce populations.

Ecological genetics. 2017;15(2):11-20
Influence of PDI gene overexpression on heterological proteins production in yeast Pichia pastoris
Tsygankov M.A., Padkina M.V.

Summary: Background. The yeast Pichia pastoris is used for synthesis of recombinant secretory proteins. Overexpression of assistant genes, coding proteins involved in secretion, is one of approaches to improve the production of target protein. PpPDI gene encodes P. pastoris yeast protein disulfide isomerase (Pdi). The aim of our study was to evaluate the effect of Pdi overproduction on recombinant interferons (human interferon-alfa16 and chicken interferon-gamma) production.

Materials and Methods. PpPDI gene was cloned under the control of the AOX1 gene promoter in plasmid pPICZαA. Primers for AJ302014.1 nucleotide sequence of NCBI data base were used for PpPDI gene cloning. The chromosomal DNA of the GS115 strain was used as a template. To generate strains with PpPDI gene overexpression we used a previously obtained strains producing human interferon-alfa16 and chicken interferon-gamma. Yeast transformation was performed by electroporation. Cultivation was performed using single and two-stage strategies in standard media containing methanol as the sole carbon source to induce the AOX1 gene promoter.

Results. We obtained interferon-producing strains with PpPDI gene overexpression. Over-expression of the PpPDI gene in yeast P. pastoris increases the production of interferon-alfa16, a protein containing disulfide bonds, regardless of the mode of cultivation. Effect of PpPDI gene over-expression on the production of interferon-gamma – the protein without disulfide bonds, depends on cultivation mode.

Conclusion. PpPDI gene overexpression can be used to enhance the production of interferons and other proteins that contain disulfide bonds. Effect of PpPDI gene overexpression on recombinant proteins without disulfide bonds may depend on cultivation procedure.

Ecological genetics. 2017;15(2):21-30
Hybridization between different lineages of pidschian-like whitefishes (Coregonus lavaretus pidschian, coregonidae) in water bodies of Southern Siberia
Bochkarev N.A., Zuykova E.I., Solovyov M.M.

Summary: Background. The purpose of this work was studying of interaction between morphologically similar forms of the pidcshian-like whitefishes inhabiting restricted territory of “riverine-lacustrine” system situated in the Bolshoi Abakan Rriver basin.

Material and methods. In the morphological analysis were used 141 whitefish individuals from Lake Karakul and 26 – from the Bolshoi Abakan River. In the genetic analysis 11 and 26 whitefish individuals were used respectively. For them the sequences of a gene 16S-ND1 of mitochondrial DNA (mtDNA) and ITS1 of nuclear DNA (nDNA) were defined.

Results. The whitefish populations clear differ on the gill raker numbers on the first branchial arch, growth. The analysis of the 16S-ND1 mtDNA genes variability revealed among them several divergent lineages and two lineages were revealed on the structure of ITS1 fragment of the nDNA.

Conclusions. Based on the obtained data it is proved that two genetically divergent populations of the morphologically same forms of pidcshian-like whitefishes inhabiting the “riverine-lacustrine” system (the Bolshoi Abakan River – Lake Karakul). Our results are supported a hypothesis of multiple hybridization between week differentiated populations/form of pidcshian-like whitefishes in habiting region during the postglacial period.

Ecological genetics. 2017;15(2):31-43
Physiological mechanism epistatic interaction of resistance genes to acaricides of various chemical classes in the interline hybrids of two-spotted spider mite
Sundukov O.V., Tulaeva I.A., Zubanov E.A.

Summary: Background. The presence in interline hybrids two-spotted spider mite Tetranychus urticae Koch two genes determining resistance to acaricides of various chemical classes significantly increases their sensitivity to the action of each these toxicants.

Materials and methods. The resistant and susceptible to malathion, bifenthrin and abamectin inbred lines of spider mite by disruptive selection cycles were obtained. The toxicological tests were performed by diagnostic concentrations of acaricides. The protein marker gene of resistance to malathion was determined by poliacrylamide disc-electrophoresis.

Results. The epistatic interaction of resistance genes to different acaricides is not manifestation at the stages of transcription and translation of genetic information.

Conclusion. The epistatic effect of another gene on the resistance gene to the current acaricide is a different consequence of metabolism processes encoded by each gene at the stage of phenotypic regulation.

Ecological genetics. 2017;15(2):44-49
Changes of the genetic structure in chronically irradiated scots pine populations
Kazakova E.A., Volkova P.Y., Geras’kin S.A.

Summary: Background. The vast forest areas were affected by radioactive fallout of the Chernobyl accident. Data about the long-term effects of chronic radiation exposure are insufficient, as well as knowledge about its influence on the genetic structure of populations. Study of isozyme polymorphism allows us to identify how chronic low-dose radiation exposure affects the genetic structure of plant populations and to develop new methods for assessing the biological effects of anthropogenic impact on terrestrial ecosystems.

Materials and methods. The study was conducted in six sites in the Bryansk region (4 radioactively contaminated and 2 reference ones). The method of vertical electrophoresis in PAAG was used for estimation of polymorphism of three enzymes involved in metabolic pathways. Activity of enzymes in seeds of Scots pine was assessed by spectrophotometry.

Results. The frequency of mutations in isozymes loci increased along the level of radiation exposure (7-130 mGy/year), as well as some characteristics of the genetic structure of populations. Biochemical activity of studied enzymes does not depend on the level of dose absorbed in the generative organs of pine.

Conclusions. It was found that the frequency of mutational events in loci of catabolic/anabolic enzymes and in the loci of previously studied antioxidant enzymes increases along the level of radiation exposure. The degree of the biological effect of chronic exposure was different in groups of catabolic/anabolic enzymes and antioxidant enzymes. Radiation exposure contributes to changes in the genetic structure of Scots pine populations.

Ecological genetics. 2017;15(2):50-61
Variability of cytogenetic disturbances in Lonicera caerulea (blue honeysuckle) population in an active fault zone
Boyarskikh I.G., Kulikova A.I.

Summary: Background. In active fault zones, geophysical and geochemical anomalies may have a genotoxic effect on plants growing there, as one of the factors of evolutionary transformation of plant populations.

Materials and methods. We applied a cytogenetic analysis to evaluate the genotoxic effect on a Lonicera caerulea L. (blue honeysuckle) natural population in one of the active fault zones in the Altai Mountains.

Results. We derived principal cytogenetic indices (i.e., mitotic, prophase, metaphase, anaphase, and telophase indices as well as proportion and range of abnormal mitoses) for meristematic cells of Lonicera caerulea seedlings. The increase in the mitotic activity of meristematic cells from the sites in the local fault zone is connected with the occurrence of the prophase-metaphase block to prevent consequences of an increased cell death (as a result of abnormal mitoses in these phases) and to compensate their losses by a greater number of divisions. We observed the increase in the proportion of abnormal mitoses in samples from almost all the test sites, compared with the control site. This demonstrates the increase in the genotoxic effect of geophysical and geochemical anomalies in these sites. The range of abnormal mitoses of samples from all the test sites shows the increase in the proportion of abnormalities in metaphase, compared to the control site where they can be equally found in metaphase, anaphase, and telophase.

Conclusion. The results demonstrate changes of mitotic activity, frequency of occurrence and the spectrum of mitotic anomalies in the root meristem of blue honeysuckle, which grows in conditions with contrast geophysical characteristics.

Ecological genetics. 2017;15(2):62-70

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