Том 60, № 8 (2024)

Fetal growth restriction is a complication of pregnancy that defined as the inability of the fetus to realize its genetically determined growth potential. Despite the high social and medical significance of this problem the exact pathogenesis of fetal growth restriction is not known by now. Therefore, the analysis of the molecular genetics mechanisms of this pathology within the framework of approaches using modern high-performance technologies of next generation sequencing is of undoubted interest. In this review we focused on the analysis of data obtained in studies of the fetal growth restriction genetics component. The authors of these researches used next generation sequencing technologies and carried out whole transcriptome profiling. The results of the genes expression genome-wide analysis in placental tissue allow us to identify 1430 differentially expressed genes between fetal growth restriction and normal pregnancy, of which only 1% were found in at least two studies. These differentially expressed genes are involved in the Wnt/β-catenin signaling pathway that plays an important role in cell migration, neural pattern formation and organogenesis during embryonic development. Common genes are associated with both obstetric and gynecological diseases, as well as with various somatic conditions from the groups of neurodegenerative, cardiovascular diseases and mental disorders, which probably reflects their involvement in the development of postnatal consequences of fetal growth restriction. The results of our work don‘t point only to potential molecular mechanisms and key genes underlying fetal growth restriction, but also indicate the important role of gene-gene communications in this pathology implementation: about 30% of all identified differentially expressed genes products interact with each other within the same gene network. In general, genome-wide RNA sequencing combined with the analysis of protein-protein interactions represents a promising direction in research on the development and functioning of the placenta, as well as the identification of placental insufficiency diseases genetics mechanisms, including fetal growth restriction.

The production of microorganisms that produce biologically active compounds for agriculture, the chemical, veterinary and pharmaceutical industries as well as for environmental protection continues to be an important direction of microbial biotechnology. One of the most effective approaches to the production of producers is chemical mutagenesis, which, in combination with the right breeding strategy, makes it possible to obtain highly productive strains. A significant disadvantage of chemical mutagenesis is the large number of induced mutations in the genomes of mutant strains, which makes it difficult to identify genes and, accordingly, biosynthetic pathways involved in the production of a given compound. The solution to this problem is modern technologies of genome sequencing and analysis, which make it possible to identify new genes and unknown biochemical pathways involved in the formation of biologically active compounds. The aim of the work was to analyse the genome of the mutant strain B-162/2 of the bacterium Pseudomonas chlororaphis subsp. aurantiaca, which is capable of increased production of biologically active compounds of the phenazine series and is resistant to hydrogen peroxide. When analysing the genome of strain B-162/2 in full size, 6482 coding sequences and 64 coding RNA sequences were identified. Comparison of the genome of the B-162/2 strain with the genome of the wild type B-162 allowed the identification of 39 mutations, 5 of which are localised in intergenic regions, and 34 affected coding sequences. Of the mutations detected, 14 led to a radical amino acid substitution in the proteins and 2 led to the formation of premature stop codons (methyl group sensor and MFS-type transporter). Several substitutions with high values of the Grantham coefficient were found, which could possibly lead to a change in the activity of the proteins concerned. The presence of three regions with phage genes in the genome of the B-162/2 strain was detected.

The inter- and intra-specific structure of genetic variability was studied using 18 microsatellite loci (nSSRs) in closely related roburoid oaks in the Crimean-Caucasian region.The seven most widespread Quercus taxa in the region were studied in 29 morphologically pure populations from different parts of the North Caucasus, Transcaucasia, Crimea and northeastern Europe. Most taxa were studied using nSSR markers for the first time. Among the 492 trees studied, Bayesian clustering method implemented in STRUCTURE identified clusters corresponding to the pedunculate oak Quercus robur, the Hartwiss oak Q. hartwissiana, the Caucasian oak Q. macranthera, the downy oak Q. pubescens and three subspecies of sessile oak: Q. petraea ssp. petraea, Q. petraea ssp. iberica, Q. petraea ssp. medwediewii. Geographic structure was identified within Q. robur, Q. pubescens and Q. p. ssp. petraea. The 18 nSSR loci used are efficient in the taxonomic assignment of individuals, and identifying hybrids. The close relationship between the “long-pedunculate” roburoid oaks (Q. robur and Q. hartwissiana) is shown, with a greater difference from other species. For one of the subspecies of sessile oak, widespread in the North Caucasus and Crimea Q. petraea ssp. medwediewii (syn. Q. calcarea), or limestone oak, significant differences from other taxa were found, reaching the species level. The assumption of a possible hybrid origin of this taxon as a result of hybridization of Q. petraea and Q. pubescens is not confirmed by genetic analysis. The other two subspecies of Q. petraea (Q. p. ssp. petraea and Georgian oak Q. p. ssp. iberica) are differentiated to a lesser extent and are related to each other, which confirms the legitimacy of distinguishing two geographically isolated taxa at the rank of subspecies. The highest variability was observed in Q. pubescens (H_e = 0.777). In Q. p. ssp. medwediewii variability was lower than in other widespread taxa (H_e = 0.652), and was approximately at the level of variability of Q. hartwissiana (H_e = 0.633) and Q. macranthera (H_e = 0.659). Clear differentiation of taxa by nuclear markers shows the limited introgression in closely related oak species in the Caucasus and Crimea. The identified genetic clusters can be used as reference groups for further population genetic studies of oaks in the Crimean-Caucasian region.

Marker-based selection on reindeer meat productivity is in the early stages of development, which requires the study of variability in candidate genes for meat productivity. We chose the calpastatin and androgen receptor genes as such genes to study. Polymorphisms and indels in the androgen receptor gene have been associated with height and weight characteristics in different domesticated animal species. Variation in the region of the calpastatin CAST gene, according to the results of many studies, has been associated with meat quality and meat productivity of livestock. Principal component analysis of CAST variability has grouped together wild and domestic deer from Yakutia, as well as wild and domestic deer from the Amur region, which implies gene flow between local breeds of domesticated deer and wild populations. Moreover, in the case of three microsatellite loci found in this study in the intron of the androgen receptor, principal component analysis separated wild and domestic deer.

Using the next-generation sequencing (NGS) technology, the nucleotide polymorphism in a fragment of the ND4 gene encoding NADH dehydrogenase subunit 4 was studied in 14 samples from three populations of the bird cherry-oat aphid (Rhopalosiphum padi L.) and the range of nucleotide polymorphism was determined. The insects were collected in 2021 and 2022 in the North-West of Russia (in the vicinity of St. Petersburg) and in the northern Caucasus (Krasnodar Territory and Dagestan). Mitochondrial DNA haplotypes were identified, which have 97.95–99.80% sequence identity with the reference GenBank accession number KT447631.1. The level of intraspecific polymorphism of a 438 bp ND4 gene fragment in Rh. padi varied from 0.2 to 4.3%. In the two-year experiments, 33 polymorphic sites (17 transitions and 16 transversions) were found in the ND4 sequences, which made it possible to identify 30 mitochondrial DNA haplotypes. The Rh. padi populations collected simultaneously on different host plants or at different times on bird cherry (spring) and cereals (summer) differed in the proportion of the main haplotype, as well as in the composition of unique minor haplotypes. Analysis of the ratio of mitochondrial DNA haplotypes suggests the important role of the host plant genotype in the formation of the structure of Rh. padi populations.

The thermoreceptor gene TRPM8 (rs7593557) was analyzed in 15 population samples living in different regions of Northern Asia, the Altai-Sayan Highlands and Canada. High frequencies of a rare genotype and allele in the populations of the Altai-Sayan region and Western Siberia were found among the Telengits (22.2 and 41.7% respectively), low among the Siberian Tatars (3.2 and 18.5% respectively), and among the peoples of northern Siberia and the Far East high frequency was found among the Nanais (22.2 and 42.6% respectively), and a low frequency among the Yakuts (1.1 and 10.9% respectively). No deviations from Hardy-Weinberg equilibrium were found. Trans-association of TRPV1, TRPA1 and TRPM8 gene polymorphisms was carried out using the regression analysis method in 14 populations. Of all three pairs, only one, TRPV1/TRPM8, showed a positive correlation (0.55, df = 13, P-value – 0.032). In pairs TRPV1/TRPA and TRPA1/TRPM8, a negative correlation was revealed (–0.545, df = 13, P-value – 0.048) and (–0.46, df = 13, P-value – 0.097) respectively. Our data indicate that the studied polymorphisms of the TRPV1 and TRPM8 genes are correlated with each other and negatively correlated with TRPA. The results obtained may indicate the coevolution of these genes. It was previously shown that TRPV1, as well as TRPM8, was detected with high frequency in the Nanai population. It is likely that these two mutations came simultaneously from East Asia and spread throughout Russia, which explains the fact that they are positively correlated with each other.

Using phylogenetic analysis of mitochondrial whole genome nucleotide sequences (mtDNA), allowing the study of genetic changes over many generations, a spectrum of nucleotide substitutions (along the L-strand of mtDNA) was reconstructed in European populations. The spectra of mtDNA nucleotide substitutions observed in a heteroplasmic state (at ≥1% and ≥5% levels) in first generation children were also analyzed. It was found that the spectra of nucleotide substitutions reconstructed over one and many generations do not differ practically in the main parameters: the distribution of pyrimidine and purine substitutions (with predominance of transitions T>C), the ratio of the number of transitions and transversions. Analysis of the phylogenetic tree of mtDNA haplotypes in Europeans clearly revealed the influence of negative (purifying) selection on mitochondrial gene pools. It is suggested that the selective processes guiding the mtDNA evolution in one and many generations are of a similar nature, i.e., caused by negative selection. The problem of how mutations occur and spread in mitochondria of germ line cells is discussed.

Triticum militinae Zhuk. et Migusch. – tetraploid wheat with the GAA genome, considered a natural naked mutant of T. timopheevii Zhuk. Previously, experiments were conducted for this wheat to study the karyotype and crossing characteristics. To clarify its origin and relationship with other representatives of the wheat family, analysis of the chloroplast genome of T. militinae, which has previously remained unstudied, is of great interest. The sucrose gradient method was used to isolate chloroplast DNA from leaves. Sequencing was performed using the FASTASeq 300 Sequencing Kit V1.0 100 M reads/flow cell on a Genolab M sequencer (GeneMind, China). For the first time, we sequenced and annotated the complete chloroplast genome of T. militinae with a size of 135898 bp. In the structure of the plastid genome there are a pair of inverted repeats with a size of 21552 bp each, the region of a small single copy (SSC) – 12791 bp and the region of a large single copy (LSC) – 80003 bp. As part of the chloroplast genome of T. militinae 132 structural genes were found, of which 85 protein-coding genes, 31 tRNAs and 4 rRNA genes.