The objects of investigation were leaves and inflorescences of Alchemilla subcrenata and Veronica chamaedry, growing on the left bank at 700 m from the edge of Lake Baikal. Some small differences in the quantity and quality of phenolic compounds in the leaves of one species even selected at one time were found. At the same time, differences in the quantity and quality of phenolic compounds in the different times of the day were even more pronounced. In the inflorescences of Alchemilla subcrenata the dynamics of flavonoids has been characterized by more stable composition than in its leaves. For Veronica 's tissues and organs more variety of seasonal cocktail of phenolic compounds than in the Alchemilla subcrenata was detected . Expect a variety of phenolic compounds in inflorescences of Veronica seasonal cocktail was higher than in the leaves.
Growth Behavior of Phytopathogen Clavibacter michiganensis ssp. sepedonicus Treated with Selenium Biocomposites of Mushroom Origin
Perfileva A.I., Tsivileva O.M., Koftin O.V.
The results of studying the effect of biologically obtained selenium nanocomposites on the bacterium Clavibacter michiganensis ssp. sepedonicus ( Cms ) are presented. Cms is a Gram-positive bacterium, which causes one of the most dangerous potato diseases, ring rot. The effective alongside ecologically safe methods for combating Cms are lacking. As the agents feasible for use in this purpose, we examined the selenium nanocomposites obtained from the macrobasidiomycetes' submerged cultures. For exploring the bionanocomposites effect on Cms , the methods of agar well diffusion, the suspension turbidity measurement, and the colony-forming units count were applied. The results showed that all the nanocomposites under study lowered the bacterial suspension's absorption values compared to the reference specimen, that testified to the observation of bacteriostatic effect of the agents tested. The suppression action of nanocompostes was elucidated by means of both agar well diffusion assay and colony-forming units count. Thus, the results obtained demonstrate the occurrence of bacteriostatic and bactericidal effects of the substances under study, and favor the supposition on advisability of further research into the selenium nanocomposites as the agents for agricultural recovery from the bacterial pathogens.
Dynamics of Vitamins and Phenols of Alchemilla subcrenata by Diurnal Variation of Temperature in October
Zhivetev M.A., Rudikovskaya E.G., Dudareva L.V., Graskova I.A., Voinikov V.K.
Round-the-clock dynamics of phenols and vitamins В6, С, РР in the leaves of Alchemilla subcrenata with a glance of thermal change was shown. The maximum of content of phenols and vitamin С was observed at 6 a.m. Dynamics of vitamins В6 and PP, actively participating in the exchange of proteins and amino acids, also is subject to fluctuation during the day. More higher content of these coenzymes are the coolest time of the day. To confirm the possible involvement of vitamins В6 and PP in the biosynthesis of proteins were examined stress proteins in leaves of Alchemilla subcrenata during those same days. Heat-shock protein HSP 17.6 was detected only a day at its maximum daily temperatures and сold regulated protein COR14b was detected in the morning, when the temperature was minimal.
Changes in Composition of Vacuolar Membrane Lipid Fatty Acids Under Osmotic Stress
Ozolina N.V., Nesterkina I.S., Dudareva L.V., Nurminsky V.N., Gurina V.V.
The investigation of influence of different types of osmotic stress (hypo- and hyperosmotic) on the composition of vacuolar membrane lipid fatty acids of red beet root ( Beta vulgaris L., variety Bordeaux) was conducted. Noteworthy, the plenty intensive stress impact didn't cause the conventional nonspecific reaction bound up with the increase of the unsaturated fatty acid content regulating membrane microviscosity. This feature apparently can be explained with the location and biochemical characteristics of the scrutinized membrane. Some reliable increases of the content of the short-chain saturated fatty acids (C14:0+C15:0) under hyper osmotic stress and the content of the long-chain saturated fatty acids (C22:0+C23:0) under both types of osmotic stress were found. It was proposed that the revealed changes in the content of saturated fatty acids of red beet root vacuolar membrane lipids were specific, and corresponding fatty acids were the most sensitive to osmotic stress impact.
The Composition of Fatty Acids of Pinus sylvestris L. of Olkha Village Surrounding Forests
Romanova I.M., Sеmenova N.V., Zhivetev M.A., Graskova I.A.
Palmitic, linoleic and α-linolenic acids are the major fatty acids during the study. Analyzing seasonal dynamics derived fatty acids revealed that for acids with 18 carbon atoms are characterized by virtually similar dynamics during the year in all years. Investigation of composition of fatty acids in the needles of different ages throughout the growing season revealed that in different periods of vegetation dominated by those or other fatty acids.
Injections of Encapsulated pH Sensor SNARF-1 do not Induce Apparent Stress Reaction in Larvae of Endemic Baikal Caddisflies Baicalina thamastoides
Gurkov A.N., Belousova I.A., Shchapova E.P., Baduev B.K., Vereshchagina K.P., Timofeyev M.A.
In the present study possibility of stress reaction of larvae of caddisflies Baicalina thamastoides Martynov, 1914 to injection of encapsulated sensors has been assessed. Activities of superoxide dismutase, non-specific esterases and glutathione S-transferase, which play important roles in functioning of organism defense systems, were used as biomarkers. Enzyme activities were measured after injections of empty microcapsules and microcapsules with fluorescent pH sensor SNARF-1, as well as after inlections of physiological saline solution and solution of the fluorescent sensor. Obtained results argue for nontoxicity and no apparent stress reaction of antioxidant and detoxification systems of B. thamastoides larvae to both types of microcapsules.
Glutathione Reductase of Vacuole. Comparison of Glutathione Reductase Activity of Vacuole and Tissue Extract of Red Beet Root (Beta vulgaris L.)
Pradedova E.V., Nimaeva O.D., Salyaev R.K.
Glutathione reductase (GR, EC 126.96.36.199) is the enzyme that reduces oxidized glutathione (GSSG) and thus regulates the redox state of glutathione (GSH/GSSG). GR has been studied in most plants. This enzyme has been identified in chloroplasts and cytosol, so these cellular compartments are considered to be the main place of the enzyme localization. In the same time, just a little is known about GR vacuoles. There are no conclusive evidences to prove the presence or absence of this enzyme in the vacuoles. GR activity was found in the vacuoles of red beet root cells ( Beta vulgaris L.). The level of activity, the optimum pH and isoenzyme composition of GR were compared in the vacuoles and tissue extract of beet root. Vacuolar GR activity was quite high, it was 1.5-2 times higher than the activity of the tissue extract. Enzyme pH optimum of all the objects were identical. pH-optimum depend on the pyridine nucleotide nature: pH 7.0-8.0 was an optimal range with NADPH; pH 5.0 - with NADH. GR activity of the vacuoles and tissue extracts decreased in the presence of a noncompetitive inhibitor 1-chloro-2.4-dinitrobenzene (CDNB), indicating the specificity of this enzymatic reaction. Two bands with glutathione reductase activity have been identified in the vacuoles and tissue extracts using zymography method to determine the enzymatic activity in PAAG after electrophoresis of proteins. Belonging to the GR isoforms of these bands was confirmed by enzyme immunoassay (Western blotting). The electric mobility of isoforms of the study objects did not differ significantly. It is concluded that the biochemical characteristics of vacuolar glutathione reductase were substantially identical to the biochemical characteristics of other localization GR.
Preliminary Analysis of Hemocyanins in Hemolymph Plasma of Baikal Endemic Amphipods
Bedulina D.S., Gurkov A.N., Baduev B.K., Borvinskaya E.V., Dimova M.D., Timofeyev M.A.
Two-dimensional electrophoresis of plasma hemolymph proteins of several endemic Baikal amphipod species showed that, as expected, hemocyanin is the main major protein in plasma of these species. Isoforms of hemocyanins form on the gels maximum 24 separate spots with slightly different molecular weight and vary by their isoelectric points. Analysis of transcript sequences of hemocyanin of Baikal amphipod Eulimnogammarus verrucosus revealed a broad heterogeneity of hemocyanin isoforms, which were divided into 2 distinct groups on their amino acid sequence and have different calculated molecular weight and isoelectric points. The β-type of hemocyanin has been described for percarid crustaceans for the first time.
Determination of Glutathione and Its Redox Status in Isolated Vacuoles of Red Beetroot Cells
Pradedova E.V., Nimaeva O.D., Putilina T.E., Semenova N.V., Sobenin A.M., Salyaev R.K.
The glutathione of the red beetroot vacuoles ( Beta vulgaris L.) was measured using three well-known methods: the spectrofluorimetric method with orthophthalic aldehyde (OPT); the spectrophotometric method with 5.5'-dithiobis-2-nitrobenzoic acid (DTNB); the high-performance liquid chromatography (HPLC). The content of reduced (GSH) and oxidized glutathione (GSSG) differed depending on the research method. With OPT the concentration of glutathione was: GSH - 0.059 µmol /mg protein; GSSG - 0.019 µmol/mg protein and total glutathione (GSHtotal) - 0.097 µmol/mg protein. In the case of determining with DTNB the concentration of glutathione was: GSH - 0.091 µmol/mg protein; GSSG - 0.031 µmol/mg protein; GSHtotal - 0.153 µmol/mg protein. HPLC-defined concentration of glutathione was lower: GSH - 0.039 µmol/mg protein; GSSG - 0.007 µmol/mg protein; GSHtotal - 0.053 µmol/mg protein. Redox ratio of GSH/GSSG was also dependent on the method of determination: with OPT - 3.11; with DTNB - 2.96 and HPLC - 5.57. Redox ratio of glutathione in vacuoles was much lower than the tissue extracts of red beetroot, which, depending on the method of determination, was: 7.23, 7.16 and 9.22. The results showed the vacuoles of red beetroot parenchyma cells contain glutathione. Despite the low value of the redox ratio GSH/GSSG, in vacuoles the pool of reduced glutathione prevailed over the pool of oxidized glutathione.
Effect of Photosynthetic Electron Transport Inhibition in vivo on the Chloroplast Genes Transcription in Arabidopsis
Yakovleva T.V., Zubo Y.O., Tarasenko V.I., Garnik E.Yu., Borner T., Konstantinov Yu.M.
Transcription of both nuclear and plastid genes encoding components of photosynthetic apparatus is subjected to redox regulation. The origin of redox signals mediating this regulation is chloroplast electron transport chain (ETC) itself. Up to date the effects of redox state of individual ETC components on chloroplast genes transcription rate was demonstrated only for some selected genes and only in isolated chloroplasts. In the present work ETC inhibitors was used for in vivo modulations in plastoquinone pool redox state, and run-on transcription approach was used to evaluate the plastid genes transcription rate. We have demonstrated that plastoquinone redox state has impact on transcription rate of wide range of Arabidopsis chloroplast genes. Treatment by DCMU (plastoquinone pool is oxidized) lead to increase of transcription rate of the majority of plastid genes studied. Treatment by DBMIB (plastoquinone pool is reduced) lead to decrease of plastid genes transcription rate. Simultaneous treatment by one of photosynthesis inhibitors and KCN (mitochondrial respiratory complex IV inhibitor) lead to decrease of transcription rate of plastid genes in most cases. Our results suggest probable participation of mitochondria in the redox regulation of plastid genes expression in Arabidopsis .