No abstract

Fifty-three species of perennial grasses in the genus Elymus L. (Poaceae), which are widespread in Russia, are generally assumed to have three haplome combinations: StH, StY and StHY. The StH-genome species, endemic to Russia, remain the least studied. R. Mason-Gamer and co-authors have previously shown in a series of studies that a molecular phylogenetic analysis of the low-copy gene waxy (GBSS1) sequences significantly complements cytogenetic data on the genomic constitution and evolutionary relationships among both North American and Asian species of the genus Elymus. To determine the species’ genomic constitution and to evaluate the level of phylogenetic differentiation, we examined the GBSS1 gene in 18 species of Elymus from Siberia and the Russian Far East, including the fol- lowing 14 endemics: E. charkeviczii, E. jacutensis, E. kamczadalorum, E. komarovii, E. kronokensis, E. lenensis, E. macrourus, E. margaritae, E. subfibrosus, E. sajanensis, E. transbaicalensis, E. peschkovae, E. uralensis, and E. viridiglumis.PCR amplification products of GBSS1 gene fragments (including exons 9–14) were cloned and 6–8 clones per accession were sequenced. It appears that all the species studied have St and H subgenomic gene variations. The most significant differences between the subgenomic variants St and H were found in intron 13. The H subgenome contains a 21-bp-long deletion in intron 13 in all Elymus genotypes, probably derived from a common ancestor of the H and P genomes. Instead of this deletion, all St subgenomes have a relatively conservative sequence similar to that of the genus Pseudoroegneria, whose ancestor is considered to be the donor of the modern St subgenome for all Elymus species. Cluster phylogenetic analysis revealed differentiation in St and H subgenome sequences into two evolutionary variants: St1 vs. St2 and H1 vs. H2, respectively. Variants of the St and H subgenomes were found homologous to various modern species of the ancestral genera Pseudoroegneria and Hordeum: St1 to P. strigosa, St2 to P. spicata, H1 to H. jubatum, and H2 to H. californicum. The details of the relationships between Russian and North American species of the genus, as well as a number of microevolutionary interconnections in the group of boreal endemic species of Siberia and the Russian Far East were revealed. The new results obtained here are essential for the development of a phylogenetically oriented taxonomic system for the genus Elymus.

The hairy root culture of halophyte was obtained by transforming seedlings' primary leaves with a wild strain of 15834 SWISS (transformation frequency was 78.7%). The roots' transgenic status was confirmed by PCR analysis using -,-, -,-specific primers. There was a significant increase in the biosynthesis of flavonoids, hydroxycinnamic acids, pectins, protopectins and saponins in hairy roots as compared to intact plant roots. The сatechin content in ethanol extracts of transformed roots' exceeded this factor 3.8 times in plant roots, but the compound composition of catechin nature in the latter was more diverse. The high antiviral activity of hairy root ethanol extracts against influenza virus subtypes A(H5N1), A(H3N2) was revealed, which manifested itself in the effective inhibition of infectivity both in vitroin MDCK cell culture-and in vivo increasing survival of mice infected with them.

A comparative study was made of the sequence of the GBSS1 gene fragment in accessions of closely related species Elymus margaritae, E. komarovii, and E. transbaicalensis from Siberia. Nucleotide sequences of the gene were determined for St subgenomes in accessions of E. margaritae and E. komarovii from classical habitats. The StH-genomic constitution was confirmed, and microevolutionary relationships between species were evaluated by constructing the NJ dendrogram. It was shown that variants of the St subgenomes in accessions E. margaritae GUK-1009 and E. komarovii AUK-9803 belong to the North American ancestral line St2 of the genus Pseudoroegneria, in contrast to accessions E. margaritae AUK-0650 and E. komarovii GAR-0501. The latter belong to the Asian branch of St1, together with variants of subgenomes in the species E. gmelinii and E. pendulinus. That is, according to the differentiation levels of the St subgenome, accession E. margaritae GUK-1009 (holotype) is significantly distanted from the accession AUK-0650 (paratype), which in turn is close to accessions of E. komarovii and E. transbaicalensis from East Sayan. According to levels of differentiation of the H subgenome, the studied species did not show noticeable differences.