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Vol 52, No 9 (2016)

Reviews and Theoretical Articles

Breeding of the Russian sable: Stages of industrial domestication and genetic variability

Kashtanov S.N., Sulimova G.E., Shevyrkov V.L., Svishcheva G.R.

Abstract

Creating farms for sable breeding was associated with the commercial destruction of natural populations and, consequently, the overall decline in the species number. The gene pool of the first farm-bred sable population in Russia, established in the vicinity of Moscow (“Pushkinskiy” fur farm), was formed by crossing of animals removed from nine natural populations. In the first eight years of farm operation, approximately one thousand animals were used for sable breeding; some of these animals were able to adapt to the farm management and, subsequently, to the selection for a number of quantitative traits in the period of industrial domestication. It took about ten years for breeders to work out the breeding and selection technologies, which became successfully employed in the established affiliated sable breeding farms. The main achievement in sable breeding over the 85-year historical period of breeding in Russia is the creation of two unique breeds, black sable (1969) and Saltykovskaya 1 (2007). In general, industrial domestication in fur farming and the subsequent breeding works made the fur of many species (mink, fox, Arctic fox) obtained from natural populations uncompetitive, which undoubtedly reduced the hunting interest in the animals living in the wild. Consequently, hunting for fur-bearing animals of most species decreased and has only local importance. Owing to the specific features of sable biology, the fur of farm-bred animals cannot yet completely replace the furs obtained by hunting; however, the farm-bred sable population is constantly growing. This review presents the results of the analysis of the level of genetic variability in natural and farm populations at nuclear and mitochondrial loci. The comparative analysis makes it possible to estimate the loss of genetic diversity upon the species adaptation to the new conditions of existence.

Russian Journal of Genetics. 2016;52(9):889-898
pages 889-898 views

Genetics of Microorganisms

Analysis of complete sequence of cryptic plasmid pTP33 from Yersinia pestis isolated in Tuva natural focus of plague

Afanas’ev M.V., Balakhonov S.V., Tokmakova E.G., Polovinkina V.S., Sidorova E.A., Sinkov V.V.

Abstract

This paper studies a full nucleotide sequence of cryptic plasmid pTP33, which was isolated from the typical plague strain of the Tuvinian natural focus, Yersinia pestis I-2638. Sequencing was carried out using the 454 GS Junior platform (Roche). In analysis using the software package GS De Novo Assembler v. 2.7 (Roche) and the algorithm Newbler v. 2.7, 1855 nucleotide reads, which contained 1101246 nucleotides, were assembled to a contig of 33 978 bp. The GC content of the obtained nucleotide sequence was 50.25%. During annotation, we found 56 open reading frames. Homologs of the predicted reading frames were sought in the BLAST databases. We detected 22 reading frames coding hypothetical proteins, 23 frames coding phagerelated proteins, and 11 frames coding proteins with known functions, including toxin–antitoxin system YefM-YoeB, nucleic acids and polysaccharides metabolism proteins (exopolysaccharide production protein ExoZ, exodeoxyribonuclease VIII), and replication proteins (ParA). Some predicted pTP33 proteins were found to be homologs (from 45 to 75%) with sequences of phage-related proteins of certain microorganisms—endosymbionts of insects (Sodalis glossinidius) and endosymbionts of entomopathogenic nematodes (Photorhabdus luminescens, P. asymbiotica, Xenorhabdus bovienii).

Russian Journal of Genetics. 2016;52(9):899-906
pages 899-906 views

Lactobacillus fermentum 90 TC-4 taxonomic status confirmation using whole genome sequencing and MALDI TOF mass spectrum

Belova I.V., Tochilina A.G., Solovyeva I.V., Efimov E.I., Gorlova I.S., Ivanova T.P., Zhirnov V.A.

Abstract

With the use of whole genome sequencing, the taxonomic status of Lactobacillus fermentum 90 TC-4 strain from Russian collections were studied. Complex analysis of phenotypical and genetic properties was conducted using phenotypic and molecular genetic methods. The main characteristics of the genome and biochemical activity profile of the strain were determined. A comparative analysis of the mass spectrum of ribosomal proteins of the strain, its biochemical properties, a fragment of 16S rRNA gene sequencing, and the entire genome revealed that the present strain belongs to the species L. fermentum, confirming its taxonomic status in accordance with modern taxonomy.

Russian Journal of Genetics. 2016;52(9):907-913
pages 907-913 views

Genome structure and origin of nontoxigenic strains of Vibrio cholerae of El Tor biovar with different epidemiological significance

Smirnova N.I., Kul’shan’ T.A., Baranikhina E.Y., Krasnov Y.M., Agafonov D.A., Kutyrev V.V.

Abstract

Intraspecies genetic differentiation of nontoxigenic strains of Vibrio cholerae of El Tor biovar containing one of the key pathogenicity genes, tcpA, is studied along with the phylogenetic relationships between these strains and toxigenic isolates. Comparative analysis of the whole genome nucleotide sequences demonstrates for the first time that ctxAtcpA+ strains vary considerably and can be clustered into two separate groups, the CTXφ–RS1φ+VPI+VSP+/CTXφ–RS1φ–VPI+VSP+ isolates and the CTXφ–RS1φ–VPI+VSP isolates, differing in their epidemiological significance. In the course of model experiments, it is established that nontoxigenic potentially epidemic CTXφ–RS1φ+VPI+VSP+/CTXφ–RS1φ–VPI+VSP+ isolates are derivatives of toxigenic strains. The results of whole genome SNP analysis of 35 Vibrio cholerae strains confirm these data and indicate genetic remoteness of nontoxigenic CTXφ–RS1φ–VPI+VSP strains both from the potentially epidemic strains and from the toxigenic isolates. It is found that the genomes of the CTXφ–RS1φ–VPI+VSP strains contain unique SNPs which are characteristic of them alone. The new data on the structure of the genome of nontoxigenic strains with different epidemiological significance may be further used for their genetic differentiation.

Russian Journal of Genetics. 2016;52(9):914-925
pages 914-925 views

Plant Genetics

Disequilibrium distribution of genotypes in the pairs of isozyme loci in the pine populations of the forest-steppe regions of Siberia

Tikhonova I.V., Ekart A.K., Kravchenko A.N., Zatsepina K.G.

Abstract

We performed an analysis of linkage disequilibrium of alleles of 15 allozyme loci in 35 populations of Scots pine in the Siberian part of the range between 50° and 56° N and 79° and 107° E. We found a significant deviation in the distribution of genotypes of 1–9 pairs of loci in the investigated samples. We established correlations between frequencies of certain pairs of linked loci with ecological conditions and geographical coordinates of habitats of the pine populations. We present the results of comparative analysis of the differences in certain morphological features of trees with significantly predominant genotypes for pairs of linked loci in three isolated populations: in the south of Krasnoyarsk krai, Khakassia, and Tuva.

Russian Journal of Genetics. 2016;52(9):926-938
pages 926-938 views

Expression of plant antimicrobial peptide pro-SmAMP2 gene increases resistance of transgenic potato plants to Alternaria and Fusarium pathogens

Vetchinkina E.M., Komakhina V.V., Vysotskii D.A., Zaitsev D.V., Smirnov A.N., Babakov A.V., Komakhin R.A.

Abstract

The chickweed (Stellaria media L.) pro-SmAMP2 gene encodes the hevein-like peptides that have in vitro antimicrobial activity against certain harmful microorganisms. These peptides play an important role in protecting the chickweed plants from infection, and the pro-SmAMP2 gene was previously used to protect transgenic tobacco and Arabidopsis plants from phytopathogens. In this study, the pro-SmAMP2 gene under control of viral CaMV35S promoter or under control of its own pro-SmAMP2 promoter was transformed into cultivated potato plants of two cultivars, differing in the resistance to Alternaria: Yubiley Zhukova (resistant) and Skoroplodny (susceptible). With the help of quantitative real-time PCR, it was demonstrated that transgenic potato plants expressed the pro-SmAMP2 gene under control of both promoters at the level comparable to or exceeding the level of the potato actin gene. Assessment of the immune status of the transformants demonstrated that expression of antimicrobial peptide pro-SmAMP2 gene was able to increase the resistance to a complex of Alternaria sp. and Fusarium sp. phytopathogens only in potato plants of the Yubiley Zhukova cultivar. The possible role of the pro-SmAMP2 products in protecting potatoes from Alternaria sp. and Fusarium sp. is discussed.

Russian Journal of Genetics. 2016;52(9):939-951
pages 939-951 views

Animal Genetics

Nuclear mtDNA pseudogenes as a source of new variants of the mtDNA cytochrome b haplotypes: A case study of Siberian rubythroat Luscinia calliope (Muscicapidae, Aves)

Spiridonova L.N., Valchuk O.P., Red’kin Y.A., Kryukov A.P.

Abstract

Sequence polymorphism of the mitochondrial DNA cytochrome b gene fragment was analyzed in 21 specimens of subspecies Luscinia calliope calliope (Pallas, 1776) and two specimens of L. c. anadyrensis (Portenko, 1939). On sequence chromatograms, in 19 specimens of L. c. calliope, double peaks of heteroplasmy type in the taxon-specific positions were revealed. Moreover, two clone variants were identified. The first variant was the calliope mitochondrial cyt b gene and the second was the nuclear cyt b pseudogene, similar to the mitochondrial haplotype anadyrensis-camtschatkensis. In L. c. anadyrensis, four clone variants, represented by the mitochondrial calliope and anadyrensis-camtschatkensis cyt b genes and nuclear calliope and sachalinensis cyt b pseudogenes, were identified. Some nuclear cyt b pseudogenes were highly similar (98–99%) to the mitochondrial genes of the subspecies L. c. anadyrensis, L. c. camtschatkensis, and L. c. sachalinensis. In the same time, the majority of nuclear pseudogene sequences were characterized by a high level of polymorphism, caused by nonsynonymous substitutions (up to five substitutions per sequence), the presence of indels in some of the clones, and TAA and TGA stop codons. In our opinion, the mitochondrial haplotypes anadyrensis-camtschatkensis and sachalinensis occurred as a result of intergenomic homologous recombination. This finding provides a new insight into the colonization history of the northeastern part of the range by L. calliope, according to which populating the territory of Chukotka, Kamchatka, and Sakhalin took place at different times and along the independent pathways.

Russian Journal of Genetics. 2016;52(9):952-962
pages 952-962 views

Characterization of the Russian beef cattle breed gene pools using inter simple sequence repeat DNA analysis (ISSR analysis)

Sulimova G.E., Voronkova V.N., Perchun A.V., Gorlov I.F., Randelin A.V., Slozhenkina M.I., Zlobina E.Y.

Abstract

The gene pools of beef cattle breeds bred in Russia were characterized on the basis of inter simple sequence repeat DNA analysis (ISSR analysis). Samples of Aberdeen Angus, Kalmyk, and Kazakh Whitehead breeds from Russia, as well as of Hereford breed, hybrids of Kazakh Whitehead and Hereford breeds, and Kazakh Whitehead breed from the Republic of Kazakhstan, were examined. In the examined breeds, 27 AG-ISSR fragments were identified, 25 of which were polymorphic. The examined breeds were different both in the fragment profiles (the presence/absence of individual ISSR fragments) and in their frequencies. It was demonstrated that the hybrid animals lacked some ISSR fragments that were present with high frequencies in parental forms, suggesting considerable genome rearrangement in the hybrid animals (at the regions of microsatellite localization) in crossings of the individuals from different breeds. The level of genetic diversity in Russian beef breeds was consistent with the values typical of farmed populations (breeds). The genetic diversity parameters assessed by applying Nei’s gene diversity index and the Shannon index varied from 0.0218 to 0.0605 and from 0.0225 to 0.0819, respectively. The highest Shannon index value was detected in the Kalmyk breed (0.0837) and Kazakh Whitehead breed from Russia (0.0819), and the highest level of Nei’s gene diversity index was found in the Kalmyk breed (0.0562) and in both populations of the Kazakh Whitehead breed (0.0509 and 0.0605). The high level of genetic similarity (according to Nei) was revealed between Russian beef cattle breeds and Hereford cattle: 0.839 (for the Kazakh Whitehead breed from Russia) and 0.769 (for the Kalmyk breed).

Russian Journal of Genetics. 2016;52(9):963-968
pages 963-968 views

DNA barcoding of fishes in Irtysh River China

Yang T., Meng W., Zhang R., Gao T., Cai L., Hai S., Zhou Q.

Abstract

DNA barcoding was a molecular diagnostic method that provided rapid and accurate species identification. The 650 bp-length cytochrome c oxidase subunit I (COI) gene of 33 species in Irtysh River China was sequenced and analyzed in this study. The average intra-species, -genus, -family, and -order of Kimura two parameter (K2P) distances were 0.003, 0.060, 0.163 and 0.240, respectively. The genetic distance between genus Barbatula and Cobitis was the largest whereas that between genus Hypophthalmichthys and Aristichthys was the smallest. The neighbour-joining tree constructed by all 44 haplotypes was divided into two major clusters: Cypriniformes fishes and other fishes. A cryptic species of Barbatula barbatula was detected according to 2% genetic threshold.

Russian Journal of Genetics. 2016;52(9):969-976
pages 969-976 views

Identification of novel SNPs of ovine PRL gene and their association with milk production traits

Ozmen O., Kul S.

Abstract

Prolactin (PRL) is a lactogenic hormone that plays a significant role in milk production; its depletion in sheep provokes a severe reduction of milk secretion. Thus, PRL also could be used as a positional marker gene associated with milk production and composition traits. Therefore, the purpose of the study was to identify genotype frequencies of single nucleotide polymorphisms the intron 2 in ovine PRL gene and its possible association genotypes with milk traits in dairy sheep breeds. The genetic structures of ovine PRL gene were examined by PCR-RFLP and DNA sequencing methods in three sheep populations. Four hundred and fifty blood and milk samples were used in the study, which included 150 samples from each of Sakiz, Akkaraman and Awassi ewes respectively. As a result, PRL genotype AA showed a strong association with milk yields content, whereas the animals carrying BB genotype had a higher fat percentage value in the three sheep breeds. Haplotype analysis of the obtained sequences showed the presence of 12 haplotypes in the PRL intron 2 region. In the present study, we have reported for the first time 48 SNPs of the PRL gene for intron 2 in dairy sheep breeds. These preliminary results indicate that the identified SNPs lend themselves readily for further research regarding physiological impacts such as milk production and reproductive traits in other dairy sheep populations.

Russian Journal of Genetics. 2016;52(9):977-984
pages 977-984 views

Mathematical Models and Methods

Mathematical models in genetics

Traykov M., Trenchev I.

Abstract

In this study, we present some of the basic ideas of population genetics. The founders of population genetics are R.A. Fisher, S. Wright, and J. B.S. Haldane. They, not only developed almost all the basic theory associated with genetics, but they also initiated multiple experiments in support of their theories. One of the first significant insights, which are a result of the Hardy–Weinberg law, is Mendelian inheritance preserves genetic variation on which the natural selection acts. We will limit to simple models formulated in terms of differential equations. Some of those differential equations are nonlinear and thus emphasize issues such as the stability of the fixed points and time scales on which those equations operate. First, we consider the classic case when selection acts on diploid locus at which wу can get arbitrary number of alleles. Then, we consider summaries that include recombination and selection at multiple loci. Also, we discuss the evolution of quantitative traits. In this case, the theory is formulated in respect of directly measurable quantities. Special cases of this theory have been successfully used for many decades in plants and animals breeding.

Russian Journal of Genetics. 2016;52(9):985-992
pages 985-992 views

Short Communications

Genetic determinants of resistance of hospital-associated strains of Klebsiella pneumoniae to β-lactam antibiotics isolated in neonates

Dubodelov D.V., Lubasovskaya L.A., Shubina E.S., Mukosey I.S., Korostin D.O., Kochetkova T.O., Bogacheva N.A., Bistritskiy A.A., Gordeev A.B., Trofimov D.Y., Priputnevich T.V., Zubkov V.V.

Abstract

According to the results of analysis of whole genome sequencing, the presence of genes having resistance to β-lactam antibiotics in hospital-associated strains of Klebsiella pneumoniae was studied. The strains were isolated from neonatal intensive care units. The data obtained were compared with the results of antimicrobial susceptibility testing of isolated microorganisms. Among other strains resistant to cephalosporins, the dominance of genes of CTX-M-type extended-spectrum β-lactamases was shown. It was revealed that one of eight strains phenotypically resistant and moderately resistant to carbapenems have the blaOXA-48 carbapenemase gene.

Russian Journal of Genetics. 2016;52(9):993-998
pages 993-998 views

Mutational landscape of prostate tumors revealed by whole-exome sequencing

Gilyazova I.R., Yankina M.A., Kunsbaeva G.B., Klimentova E.A., Izmaylov A.A., Pavlov V.N., Khusnutdinova E.K.

Abstract

The results of the whole-exome DNA sequencing of eight prostate adenocarcinoma patients are presented. DNA was isolated from the peripheral blood as well as healthy and tumor prostate tissue from each patient. Bioinformatics analysis was conducted and the most significant mutations in prostate cancer patients were revealed. The obtained data could be important for understanding of the molecular mechanisms of prostate cancer pathogenesis and facilitate development of new approaches for treatment of the disease.

Russian Journal of Genetics. 2016;52(9):999-1003
pages 999-1003 views

Estimation of association of CNTN6 copy number variation with idiopathic intellectual disability

Lopatkina M.E., Kashevarova A.A., Lebedev I.N.

Abstract

Analysis of the prevalence of copy number variations of the CNTN6 gene, recently selected as a new candidate gene for intellectual disorders, was performed. Real-time PCR did not detect any change in the number of CNTN6 gene copies in a group of 200 patients with impaired intellectual development. However, taking into account our data from the previous aCGH analysis and published data, the overall frequency of microdeletions and microduplications of CNTN6 was estimated as 1: 265 (0.4%). The common phenotypic features of 40 patients with microdeletions and microduplications of CNTN6 appeared to be the autism spectrum disorders, developmental delay, intellectual disability, seizures, cognitive impairment, cardiological defects, and behavioral problems.

Russian Journal of Genetics. 2016;52(9):1004-1006
pages 1004-1006 views