Том 53, № 10 (2017)

This paper considers molecular mechanisms of DNA methylation and histone modifications in plants. The role of these epigenetic processes in plant development is discussed.

The study of RNA functions in a cell is based on the methods of labeling and detection of these molecules. In recent years, considerable progress has been made in this area, making it possible to detect RNA in amounts to one molecule with high specificity and to track the dynamics of spatial RNA distribution in the living cell. The review presents the latest developments in the field of in situ hybridization and the use of RNA aptamers and RNA-binding proteins. The prospects of using the CRISPR-Cas system for RNA visualization are discussed in detail.

It is shown by single-cell gel electrophoresis that the exposure of CBA mouse males with pheromone 2,5-dimethylpyrazine for 4 or 24 h increases DNA damage level in interphase nuclei of bone marrow cells. The results may reflect the work of a mechanism of formation of chromosomal aberrations and other mitotic disturbances, detected at the stage of ana-telophase, the frequency of which in dividing bone marrow cells increased after similar pheromonal exposure. The comparison of the damaged cell distribution types is proposed as an approach to analysis of comet assay data. The significance of the revealed effects on the immune system in the recipient organism is discussed.

Phototrophic non-sulfur bacteria (PNSB) are close relatives of cytokinin producing agrobacteria. However, the sequenced PNSB genomes do not have the ipt gene sequences involved in the cytokinin biosynthesis. In Rhodobacter sphaeroides and Rhodopseudomonas palustris transformed with ipt, we previously observed the substantial morphological and physiological changes as compared with the original strains. To find associations of these changes with the ipt function, its expression levels were studied in the transformants and the parent strains. Unexpectedly, the ipt transcripts were found in both the transformants and the original untransformed strains, although the ipt expression level was several orders of magnitude lower in the latter compared to the transformants. The ipt sequences amplified from the plasmid DNA templates of the parent PNSB strains showed that they were identical to each other but differed in four nucleotides compared to the ipt gene from the Agrobacterium tumefaciens pTiBo 542 plasmid. The in silico comparison amongst the ipt sequences from the other agrobacterial plasmids, as well as from the other PNSB, demonstrated their low identity. This is the first report on the active gene of the cytokinin biosynthesis found in Rba. sphaeroides and Rps. palustris which is homologous to the agrobacterial ipt gene.

The indigenous Russian Spangled Orloff chicken breed (Large Fowl) was for the first time studied by analyzing polymorphism in loci associated with viral diseases caused by Marek’s disease viruses (MDV), avian leukosis viruses (ALV), and avian influenza viruses. Samples for the analysis were collected at the farms of the All-Russian Poultry Research and Technological Institute (VNITIP), the All-Russian Institute of Farm Animal Genetics and Breeding (VNIIGRZh), and the Moscow Zoo. Assessed populations have a distinction in the frequency and range of allelic variants and as a consequence the frequencies of genotypes LEI0258. The highest level of polymorphism was found in microsatellite loci LEI0258 and Mx1 gene (VNIIGRZh population). The evaluation of polymorphism in genes Mxl (interferon-induced guanosine triphosphatase) and TVB (Tumor Virus В locus, cell receptors) showed, that the Mxl and TVB genotypes associated with resistance to avian influenza viruses and ALV, respectively, were not found in studied populations of Spangled Orloff chicken breed. The most homogeneous group for all studied loci is from the VNITIP population. These data are in good agreement with the results from the study of mtDNA polymorphism in the same samples. The observed polymorphism in nuclear and mitochondrial markers demonstrates that most representative population (about 700 individuals) of Spangled Orloff chicken breed from VNITIP during its long conservation breeding program has remained almost unchanged in the investigated molecular markers and is a source of valuable alleles 357 from microsatellite loci LEI0258 associated with Marek’s disease resistance. The observed frequency of the genotype 357/357 was 48%. The obtained data on the genetic homogeneity of Spangled Orloff chicken breed in all analyzed loci show the need for measures to maintain and increase the number of local groups of Orloffs with the aim of its further preservation.

On the basis of sequence analysis of the mitochondrial DNA (mtDNA) control region (CR), cytochrome b (Cytb), and cytochrome oxidase-1 (CoI) genes, the relationships of endemic species Salvelinus andriashevi Berg, 1948, represented by the only population from Lake Estikhed (Chukotka), were estimated. The data on the genealogical analysis of mtDNA haplotypes supported phylogenetic closeness of S. andriashevi and S. taranetzi. It was also demonstrated that the specimens of Chukchi charr, along with Salvelinus sp. 4 (Lake Nachikinskoe), S. krogiusae (Lake Dal’nee), S. boganidae and S. elgyticus (Lake El’gygytgyn), and S. a. erythrinus from Canada’s Northwest Territories (NWT) belonged to the Arctic group of Taranetz charr. The problem of coincidence of taxonomic differentiation of charrs of the genus Salvelinus based on morphological and genetic analyses is discussed.

The frequency and spectrum of mutations in RAS/BRAF genes were studied in rectal adenomas, carcinomas in situ, and adenocarcinomas. It is shown that the frequency of KRAS mutations decreases from adenoma to adenocarcinoma; most adenomas and carcinomas in situ are heterogeneous and consist of several subclones. Possible models of colorectal cancer pathogenesis are discussed.

The effect of strong hypomorphic mutation of the insulin-like protein DILP6 (dilp6), synthesized in the fat body, on the expression intensity of insulin-like protein DILP3 synthesized in median neurosecretory cells (MNCs) of D. melanogaster brain was examined. The intensity of DILP3 expression in the MNCs of the larvae was evaluated immunohistochemically using antibodies against DILP3 and confocal microscopy. For the first time, it was demonstrated that, in dilp6⁴¹ mutants, there was a sharp increase in DILP3 expression level. The data obtained indicate the existence of negative feedback coordinating the expression of insulin-like proteins synthesized in MNCs and peripheral tissues of Drosophila.