Vol 54, No 11 (2018)

The paradoxical effect of deletion of the Escherichia coli genes cysK and cysM encoding cysteine synthase enzymes has been studied: such cysteine auxotrophs actively degrade the excess of cysteine transported from the medium to form H₂S. We have shown that deletions of any of the known genes controlling the degradation of exogenous cysteine, including the genes aspC, mstA, cysK, cysM, tnaA, metC, and malY, as well as the newly discovered genes yciW, cyuA, cyuP, and cyuR, do not deprive the cysteine auxotrophs ΔcysK ΔcysM of the ability to degrade cysteine. Cysteine degradation in the ΔcysK ΔcysM mutant is positively regulated by the products of the cysB and cysE genes. It is significant that the ΔcysK ΔcysM mutant shows an increased transcription of the genes opposing the oxidative stress (sodA, catG, arcA, and cydD). We assume that oxidative stress in cells of the ΔcysK ΔcysM mutant is provoked by restriction of cysteine resynthesis, while cysB-dependent degradation of exogenous cysteine and generated H₂S provide protection against oxidative stress.

The nature of the formation of fertile progenies of F2 and F3 generations of wheat–rye hybrids obtained as a result of meiotic restitution using the Saratovskaya 29 variety (C29 x R) and the wheat–rye substituted line 1Rv(1A) (1Rv (1A) × R) was studied. The karyotype analysis of the F₂Triticum aestivum L. cv. Saratovskaya 29 × Secale cereale L. var. Onochoyskaya (S29 × R) hybrid using C-banding revealed 56 chromosomes; among them were 42 wheat chromosomes and 14 rye chromosomes. The karyotype of the F₂ 1Rv(1A) × R hybrid contained 46 chromosomes, of which three pairs of rye chromosomes 1R1R4R4R2RL2RL, 1R1R replaced the chromosomes 1A1A, and 2RL2RL and 4R4R were additional to the wheat chromosomes. In the F₃ generation of S29 × R hybrids, the octoploid number of chromosomes with aneuploidy of single rye and wheat chromosomes was preserved, while in the 1Rv(1A) × R hybrids the number of chromosomes varied from 42 to 49, but in most plants 2n = 46 was retained. Meiosis of the F₃ 1Rv(1A) × R hybrids was more stable than that of the S29 × R hybrids. The average number of univalents in 1Rv(1A) × R hybrids was 1.54 ± 0.17, which was significantly lower than in S29 × R hybrid progenies (2.56 ± 0.23 and 2.07 ± 0.27). The F₃ С29 × R and 1Rv(1A) × R hybrids were characterized by the variability in the seed set and 1000-grain weight. On average, С29 × R plants produced a smaller number of grains with but larger size than 1Rv(1A) × R plants, forming more grains with lower 1000-grain weight. The obtained results indicate the influence of the substitution 1Rv(1A) on the reorganization of hybrid genomes.

A significant length polymorphism (309–735 bp) and nucleotide variability were detected by the study of the intergenic spacer trnL-trnF of chloroplast DNA in Hedysarum species belonging to sections Hedysarum and Multicaulia (family Fabaceae). Among nineteen Hedysarum species analyzed, the region trnL-trnF was not investigated before for thirteen species. The trnL intron deletion was first discovered for four Hedysarum species: H. austrosibiricum, H. cisbaiсalense, H. theinum, and H. consanguineum. The chloroplast cytotype without deletion of the trnL intron was reported for the first time for H. alpinum. This paper proposes a hypothesis about the polyphyletic origin of the genus and mechanisms of the formation of the phenomena for the first time established for genus Hedysarum of heteroplasmy and chloroplast cytotype variability found in H. consanguineum and H. alpinum on the basis of the data of trnL intron labeling.

Most of eukaryotes is represented by unicellular organisms (protists) which are still poorly investigated owing to a number of methodological problems. In this study, single-cell PCR was used for the analysis of two unarmoured dinoflagellate species from Lake Baikal. DNA fragments from living and fixed Gymnodinium baicalense and Gyrodinium helveticum cells were successfully sequenced, which indicates the usefulness of the method in the study of uncultivated dinoflagellates. However, this approach is too laborious to be introduced into traditional monitoring studies. The data obtained confirmed the earlier suggestion that the Baikal population of heterotrophic species G. helveticum was wrongly described as an endemic species of Lake Baikal, Gymnodinium coeruleum. The sequences of 28S rRNA gene fragment and ITS2 region were determined for the first time for G. helveticum. Phylogenetic trees inferred from the 28S rRNA and 18S rRNA genes confirm the assignment of G. helveticum to the genus Gyrodinium. Analysis of 18S rRNA gene fragments from the GenBank database revealed a large number of unknown eukaryotes from different water reservoirs similar to G. helveticum. This is could be because of the existence of cryptic diversity within morphospecies G. helveticum, as well as the existence of yet unknown species from the genus Gyrodinium. Our analysis stimulates more intensive study of G. helveticum phylogeography in particular and the diversity of Gyrodinium in general.

Ladybird Harmonia axyridis is a widespread polyphage and forms stable populations in East Asia and spreading invasive populations in Europe and other regions with temperate and subtropical climate. A characteristic of haplotype diversity of native and invasive populations of H. axyridis at the atp6 gene is given. The characteristic differences in the composition and quantitative ratio of mitochondrial haplotypes between the western and eastern groups of populations of the native range are revealed. The genetic structure of invasive populations is uniform across the whole range in Europe, from the Atlantic coast to the Black Sea coast of the Caucasus, which is indicative of the spread of H. axyridis invasive populations in Europe from one center. In Europe, the mitochondrial gene pool of invasive populations is formed by two major haplotypes. This mitochondrial haplotype composition remains unchanged in the process of the invasion expansion. The possible role of mixed origin of invasive populations of H. axyridis in the formation of the invasive ability is discussed.

The genetic structure and differentiation level of the sable populations from the Altai mountain region and adjacent geographical regions were studied using nuclear and mitochondrial genome markers. The isolation of the sable from northwestern physicogeographical province of the Altai mountain region was confirmed. Previously, the taxonomic status of the sable from this region was determined according to morphological traits as the Martes zibellina averini Bashanov, 1943 subspecies. The results of the statistical analysis indicate the existence of two population groups in the Altai mountain region: northwestern (isolated by landscape peculiarities of the mountain region) and generated by the populations of the east and northeast of the Altai Mountains. The latter have sustainable migration links with the main area of sable of the Altai–Sayan Ecoregion and Central Siberian Plateau. The allelic diversity center of the studied part of the species area was detected.

Here, we determined basic differentially expressed genes in endometrial lesions compared to eutopic endometrium of the patients with endometriosis (EM). Endometrial biopsy and tissue sampling were performed in ten women with pelvic EM and nine controls at their mid-secretory phase. NGS of mRNA combined with bioinformatic analysis and complemented by RT-qPCR analysis were used for analysis. A list of differentially expressed genes of ectopic compared to eutopic endometrium during mid-secretary phase has been composed. Functional annotation has found profound gene expression perturbations in numerous EM pathways including extracellular matrix, complement and coagulation, as well as cell cycle-related gene family. The genes of alcohol dehydrogenase 1B (ADH1B) and fatty binding protein 4 (FABP4) were determined as the most dramatically up-regulated genes in ectopic endometrial lesions. Both genes were not reported before to be associated with EM. Several other genes including PLA2G2A were also up-regulated in ectopic EM lesions. Notably, FABP4 and ADH1B are key genes in basic metabolism and known for their function in adipose tissue. The results provide intriguing new avenue for endometriosis studies.

Major depressive disorder (MDD) is an important problem in psychophysical health and well-being of society in Russia and worldwide. In the present work, the role of apoptosis in the associative gene network of MDD was studied. The methods of prioritization were analyzed and candidate genes for further research were predicted. The quality of prioritization methods and their combinations was tested, and the most effective methods were selected. Prioritization was carried out for genes of the extrinsic apoptotic signaling pathway considering their role in the structure of the associative genetic network of MDD, since the external pathway of apoptosis is important for drug development. The methods of the ANDSystem, a computer system for reconstruction of gene networks on the basis of automatic data retrieval from scientific publications, were considered and used for gene prioritization. As a result of prioritization, some candidate genes were suggested for the further experimental study of their role in MDD pathogenesis, including CAV1, FYN, JAK2, PTEN, RAF1, RELA and SRC.

In the present work, we analyze the mutant strains of Streptomyces fradiae ATCC 19609 resistant to (33S)-azido-33-deoxyoligomycin A, obtained at a frequency of 10^–8 to 2.5 × 10^–9. Analysis of the obtained mutant strains allows them to be divided into two groups based on the level of resistance to oligomycin A. One mutant out of 15 obtained is resistant to oligomycin A at MIC of 0.1 nm/mL, and 14 mutant strains are resistant at MIC of 0.01 nm/mL of oligomycin A (MIC for the initial strain: 0.001 nm/mL). Using PCR analysis and subsequent sequencing by the Sanger method, three candidate genes the products of which are related to resistance to oligomycin A are analyzed: helicase IV, the C subunit and the A subunit of FoF1-ATP synthase. It is found that, in the genomes of all 15 mutants, there is a single nucleotide substitution (SNP) in the helicase IV gene, while in one strain most resistant to oligomycin A there is an additional SNP in the gene of the A subunit of FoF1-ATP synthase. This mutant strain will be used to obtain the mutants of resistance to oligomycin A with the aim of further sequencing of genomes and detection of biotarget of oligomycin A.