[Characterization of the Bellevalia sarmatica (Georgi) woronow populations from Volgograd oblast using molecular genetic identification].

DNA samples obtained from the populations of the Red Data Book species Bellevalia sarmatica (Georgi) Worovow from Volgograd oblast were examined using RAPD and AFLP analyses. DNA marking revealed considerable differences in the levels of interpopulation and intraspecific polymorphism of the Bellevalia. Furthermore, RAPD analysis, despite of lower levels of interpopulation polymorphism identified, made it possible to obtain clearer data on population subdivision upon statistical treatment of the results. The results of this study can be used in developing conservation strategies for maintenance of the species abundance and genetic diversity, as well as in elaborating the criteria for construction genetic collections.

Discovery of novel antimicrobial peptides with unusual cysteine motifs in dandelion Taraxacum officinale Wigg. flowers.

Three novel antimicrobial peptides designated ToAMP1, ToAMP2 and ToAMP3 were purified from Taraxacum officinale flowers. Their amino acid sequences were determined. The peptides are cationic and cysteine-rich and consist of 38, 44 and 42 amino acid residues for ToAMP1, ToAMP2 and ToAMP3, respectively. Importantly, according to cysteine motifs, the peptides are representatives of two novel previously unknown families of plant antimicrobial peptides. ToAMP1 and ToAMP2 share high sequence identity and belong to 6-Cys-containing antimicrobial peptides, while ToAMP3 is a member of a distinct 8-Cys family. The peptides were shown to display high antimicrobial activity both against fungal and bacterial pathogens, and therefore represent new promising molecules for biotechnological and medicinal applications.

[Comparative analysis of tobacco and Arabidopsis insertional mutants, transformed with equal vector constructions].

We have analyzed genetic effects of heterologous plant genes insertion on genome functioning of higher plants, belonging to different systematic groups (tobacco, Arabidopsis). Plants of different species were responding differently to the insertion of the same transgene, which is likely to be associated with the location of alien DNA insertion and could manifest in morphological changes spectrum and target gene expression level.

[Genetic monitoring of populations of Matthiola fragrans (Bunge) using RAPD and AFLP analysis].

The possibility of using RAPD and AFLP methods for genetic monitoring of populations of Matthiola fragrans (Bunge), a species included in the Red Book of the USSR, was shown for the first time. An analysis of inter- and intrapopulation and interspecies genome polymorphism was performed. Differences in the genetic structure of Matthiola populations from various geographical collection points were revealed. A simple method of performing RAPD analysis and the great number of unique markers found in each population compared with the AFLP analysis, as well as the good division of populations under statistical treatment, allow us to draw the conclusion that using the RAPD method in genetic monitoring of rare and insufficiently studied species is well founded.

[Analysis of the marker and target genes inheritance in the seed and vegetative progeny of transgenic tobacco with buckwheat serine protease inhibitor gene].

The stability of marker nptII and target gene of serine proteinase inhibitor BWI-1a (ISP) from buckwheat seeds after agrobacterial transformation in the new foreign genome environment has been investigated. Different vegetative and seed progeny of self-pollinated transgenic tobacco was studied after propagation at selective and nonselective in vitro conditions. For the majority of transgene lines among random sample of independent transgenic plants stable and coupled PCR confirmed inheritance and expression of marker and target genes has been demonstrated after prolonged microcutting and repeated regeneration cycles at unselective in vitro conditions. The most of the seed T1 progeny of the studied lines has maintained Km-resistance too. However all transgenic lines had significant disturbances at the microsporogenesis process. Two lines (C5 and C12) after 4-year microcutting lacked Km-resistance and at the same time lacked their antibacterial activity. Two other clones revealed variability in Km-resistance (line C7) and seedlings size (line C22) that might be a result of more than 1 transgene insertion or chimaeric nature of transgene plants so it require an additional study.

Use of buckwheat seed protease inhibitor gene for improvement of tobacco and potato plant resistance to biotic stress.

The possibility to use agrobacterial transformation of leaf discs to produce resistance to bacterial infections in tobacco and potato plants by introduction of a single gene encoding the serine proteinase inhibitor BWI-1a (ISP) from buckwheat seeds is shown. All studied PCR-positive transgenic plants exhibited antibacterial activity in biotests. It was shown that the presence of just a single gene of serine proteinase inhibitor provides sufficient protection at least against two bacterial phytopathogens, Pseudomonas syringae pv. tomato and Clavibacter michiganensis sbsp. michiganensis. The biotest including tobacco plant infection by the white wings butterfly in the green house has also demonstrated the existence of protective effect in transgenic tobacco plants. Significant genotypic variations in the protection efficiency were found between members of different genera of the same family (potato and tobacco) as well as between different lines of the same species. Northern blot analysis of four transgenic potato lines and three tobacco lines transformed by a vector plasmid containing the ISP gene of serine proteinases BWI-1a from buckwheat seeds has shown the presence of the expected size mRNA transcript.

[RAPD and ISSR analyses of species and populations of the genus Stachys].

Molecular analysis of the genome was performed for 14 species of the genus Stachys. RAPD and ISSR analyses of the Stachys genome revealed 574 polymorphic fragments, including genus- and species-specific markers. Based on the patterns, UPGMA and the Jacquard coefficient were used to estimate the genetic distances between Stachys species and populations and to construct dendrograms reflecting the phylogenetic relationships among the Stachys species. Molecular analysis of the Stachys genome refined the phylogenetic positions of some species and revealed synonymous species.

[Variation and morphogenetic characteristics of different Stachys species during microclonal propagation]. / Osovennosti morfogeneza i poiavlenie variatsii pri mikroklonal'nom razmnozhenii raznykh vidov stakhisa.

Morphogeneses of Stachys different species introduced in culturing in vitro have been compared. The frequency of altered forms have been demonstrated to be related to the plant genotype. All regenerants of S. sieboldii, which reproduces in vivo only vegetatively, are phenotypically normal, irrespective of the concentrations of plant growth regulators at which they have been obtained. Only changes in isozyme patterns have been observed in the regenerants grown in media containing at least 10 mg/l benzyl aminopurine (BAP); most of these changes are the absence of a particular component of the pattern. The cross-pollinating species Stachys ocymastrum, which typically reproduces by seeds, has yielded morphologically altered forms even in phytohormone-free media; its isozyme patterns often contained a new component. Analysis of the isoperoxidase patterns of regenerants of both Stachys species obtained with the use of high phytohormone concentrations has demonstrated qualitative and quantitative changes suggesting the appearance of somaclonal variants even in the course of plant regeneration directly from nodal segments, bypassing callus formation. Changes have also been found in Stachys plants regenerating from the callus tissue.

[The detection of genome polymorphism in Stachys species using RAPD]. / Ispol'zovanie RAPD analiza dlia vydeleniia genomnogo polimorfizma u predstavitelei roda Stachys.

Molecular genome analysis was for the first time carried out in the genus Stachys. RAPD analysis proved to be suitable for identifying the species-specific markers, studying the interspecific DNA polymorphism, and detecting the genetic changes that arise during in vitro culturing of Stachys sieboldii. In addition, RAPD was used for screening genetic variation in S. sieboldii regenerants obtained at various phytohormone concentrations. High cytokinin concentrations and multiple regeneration were shown to induce genetic changes detectable with RAPD patterns. High DNA polymorphism was demonstrated for two types of S. sieboldii callus cultures and for plants regenerated from a callus culture.

[Isoenzyme variants of acid peroxidase in the genus Stachys]. / Izofermentnye spektry kislykh peroksidaz u predstavitelei roda Stachys.

Isozyme patterns of acid peroxidases and their dependence on plant age, cultivation conditions, and tissue type have been investigated by electrophoresis in polyacrylamide gel in four species of the genus Stachys. The most stable peroxidase patterns have been found in the plant roots. Acid peroxidases have been shown to be species-specific, which allows their use in taxonomic studies. Cultivation in vitro and in vivo produces different isozyme patterns in various tissues of plants of various ages.

[Change in the spectra of peroxidases in regenerants of Stachys sieboldii (Miq.) as a result of hormonal and mutagenic effects]. / Izmenenie spektrov peroksidaz u regenerantov Stachys sieboldii (Miq.) vrezul'tate gormonal'nykh i mutagennykh bozdeistvii.

Nodular segments of the stem and stolons of aseptic Stachys sieboldii (Miq.) plants were cultured in Gamborg's B5 medium at a low (1-2 mg/l) or high (10 mg/l) concentration of cytokinins (6-BAP, kinetin) and produced many adventitious buds and shoots without callus formation. Several plants were regenerated from nodular segments incubated at various concentrations of 5-bromodeoxiuridine (5-BdU) for three weeks. All regenerants were morphologically normal. Peroxidase electrophoretic patterns were analyzed to determine whether somaclonal variation is associated with S. sieboldii micropropagation. The frequency of changed forms was higher with the high 6-BAP concentration and with mutagenic treatment. Changed forms were not detected in plants grown at a low cytokinin concentration, which can be used in S. sieboldii micropropagation.