Vol 18, No 4 (2020)

Background. Agrobacterium rhizogenes rol genes cause not only hairy root syndrome in plants, but also affect their secondary metabolism. There are cases of increasing of nicotine content in transgenic tobacco roots expressing rolC alone or in combination with other rol genes. In this work, we evaluated the change in the expression of nicotine synthesis genes and their regulators in response to the induction of expression of rolC.

Materials and methods. Plant material was represented by three Nicotiana tabacum genotypes: cv. Samsun and two transgenic lines, derived from this cultivar and containing rolC under dexamethasone inducible promoter: A. rhizogenes rolC (Pdex-A4rolC) and N. tabacum rolC (Pdex-trolC) correspondingly. Fluidigm Biomark RT-PCR was used for evaluation of expression of QPT1, QPT2, A622, ODC, ADC, PMT1, PMT2, PMT3, PMT4, MPO1, MPO2, BBL, MATE1, MATE2, ARF6, ERF168, ERF189, A4rolC, NtrolC, and reference gene gapdh. HPLC-MS / MS analysis was used to determine content of nicotine and its derivatives in plant tissues.

Results. Expression of PMT genes for the synthesis of the pyrrolidine ring, as well as the genes, controlling enzyme for final stages of nicotine synthesis, was higher in transgenic lines without induction of rolC expression. Regulatory genes were activated by dexamethasone in both transgenic and control lines, indicating the inapplicability of rolC dexamethasone induction for their study. The level of expression of PMT and MPO genes increased over time in transgenic dexamethasone-induced lines. Nicotine content decreased in transgenic dexamethasone-induced plants.

Conclusions. The rolC gene does not play a primary role in the regulation of nicotine synthesis genes. The mechanism of regulation of different nicotine biosynthesis genes and TFs varies.

The present and literature data showed that Meromyza flies developed on grasses from 5 tribes: Poeae, Triticeae, Bromeae, Nardeae, Arundinarieae. The preference of host plants for 25, mainly Western Palaearctic species of Meromyza flies was analyzed: 11 species developed on grasses of the tribe Poeae, 4 – on Triticeae, 9 – on grasses from different tribes, 1 species developed on bamboo. A phylogenetic tree based on the mtDNA CO1 gene locus was constructed in the BEAST for 28 species of Meromyza flies, for 8 species of Drosophila and Campiglossa pygmaea. The host plants were known for 19 species Meromyza flies out of 28 studied species. An overview of the evolution of grasses is given. By the possible time of the genus Meromyza origin (not earlier than the middle of the Miocene), based on the known evolutionary scale of Drosophila, the Pooideae grasses had already been isolated and division into tribes occured. The features of non-specialized phytophage-oligophage (except M. acuminata) confirmed by the wide spectrum of host plants have been supposed for species close to ancestral haplotypes (M. nigriseta, M. pratorum, M. saltatrix, M. variegata) or representing independent branches in their clusters (M. acuminata, M. mosquensis, M. nigriventris). The differentiation of Meromyza genus with formation of new species with narrow oligophagy or monophagy was associated with adaptation to other wild grasses following the formation and increase in the abundance of “core pooids” (Triticodae + Poodae) grasses and the spread of herbal biomes in the Miocene. Oligophages M. nigriventris, M. nigriseta, M. variegata and monophages M. acuminata, M. grandifemoris damage cereal cultivars.

We studied changes in the stability of the genome in cells of two brain regions (prefrontal cortex and hippocampus), as well as in the bone marrow of rats with a hereditary high and low thresholds of excitability of the nervous system (strains HT and LT, respectively) after prolonged exposure with emotional-pain stressor. To study the reactivity of the brain cells genome, phosphorylated histone γ-H2AX (γ-H2AX phospho Ser139) was used. The level of mitotic disturbances in bone marrow cells was also assessed. Between the animals of the control groups, there were no interstrain differences in the studied parameters. Stress exposure increases the immunoreactivity to γ-H2AX phospho Ser139 of the prefrontal cortex cells and the level of chromosomal aberrations in bone marrow cells in animals of both strains. In cells of the dentate gyrus of the hippocampus, a specific increase in immunoreactivity to γ-H2AX phospho Ser139 was revealed in rats of the low-excitable HT strain. The relationship between the reaction of cells of this zone of hippocampus to the stressor exposure with the hereditary level of excitability of the nervous system of animals is discussed.

Background. The study of the long-term effects of chronic radiation exposure on plants and animals, which are still the subject of scientific discussion, is necessary to understand the consequences of radiation accidents. After the Fukushima nuclear power plant accident, some of the young pines and spruces showed an increased frequency of apical dominance cancelling. The most probable cause of the observed morphoses is associated with damage to the apical meristem of coniferous plants by radiation in the first year of the accident, when they received the highest absorbed doses. If this hypothesis is true, then even 8 years after the accident it will be possible with high degree of probability to detect an increased level of cytogenetic abnormalities in the intercalary meristem of needles of plants from these populations.

The aim of this work was to verify this hypothesis.

Materials and methods. Five populations of Japanese red pine from territories contaminated with radionuclides as a result of the accident at the Fukushima nuclear power plant were investigated. The frequency and spectrum of cytogenetic abnormalities in the intercalary meristem of needles were determined by the ana-telophase analysis.

Results. The frequency of aberrant cells in the needles intercalary meristem of Japanese red pine from the contaminated with radionuclides territory statistically significantly exceeds the control level in all impact sites and increases along with the dose rate. Although there is no correlation between the frequency of cytogenetic abnormalities in needles and the presence of cancellation of apical dominance in plants, all pine populations from radioactively contaminated territories are characterized by an increased frequency of both cytogenetic abnormalities and morphoses associated with the cancellation of apical dominance.

Conclusion. Radiation damage to the apical meristems of conifers in the first year of the accident, when they received the highest absorbed doses, is the most likely cause of the increased frequency of cancellation of apical dominance in the studied populations of Japanese red pine from the zone affected by the accident at the Fukushima nuclear power plant.

This review article is dedicated to a relatively young, actively developing approach to biodiversity assessment – analysis of environmental DNA (or eDNA). Current views on the nature of eDNA, a brief overview of the history of this approach and methods of eDNA analysis are presented. Major research directions, utilizing eDNA techniques, and perspectives of their application to the study of biodiversity are described. Key issues in development of eDNA approach, its advantages and drawbacks are outlined.