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Evaluation of the Effect of Oxidative Stress on Roots of Red Beet ( Beta vulgaris L.)
стр.5-12
Kolesnikova E.V., Ozolina N.V., Nurminsky V.N., Nesterkina I.S., Sitneva L.A., Lapteva T.I.
In the present investigation it was studied the effect of oxidative stress on root of red beet. The degree of stress exposure was assessed at various levels: in tissues of roots (conductometric method), on isolated vacuoles (time-lapse video recording), and transport activity of the vacuolar membrane enzyme, namely V-H +-ATPase (fluorescent probe method). The obtained results allow to conclude the negative effects of oxidative stress on different levels of plant organization. There were several effects: a significant increase of outflow of electrolytes from the tissues of roots, a great decrease of half-life time of isolated vacuoles, and a 2 times reduction of transport activity of tonoplast H +-ATPase. Thus, the methods used in this study can be applied to assess the intensity of the impact of oxidative stress on plants.
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Activation of Cell Death in the Sugar Cane Suspension Culture by the Exposure to High Temperature
стр.13-24
Lyubushkina I.V., Fedyaeva A.V., Pobezhimova T.P., Stepanov A.V., Rikhvanov E.G.
The process of cell death in a sugar cane suspension culture after exposure to high temperature (45, 50, 55 and 60 °C) during 10 min has been studied. It has been revealed that treatment of cell culture at 50 °C did not cause an immediate cell death, but 50% of the cells were dying for the next 48 h. Exposure of cell culture to more high temperature (55 - 60 °C) caused a massive cell death occurred instantly after treatment. The development of cell death after the treatment at 50 °C was accompanied by the protoplast condensation, increased generation of reactive oxygen species and hyperpolarization of the mitochondrial inner membrane. Obtained results indicate on the active character of the cell death process, induced by the moderate heat shock in sugar cane suspension culture.
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Physiology and ion relations in response to salinity in Trifolium isthmocarpum Brot. and Lotus ornithopodioides L
стр.25-41
Bennani Kawtar, Bendaou Najib
Salt tolerance in Moroccan ecotypes of Trifolium isthmocarpum Brot. and Lotus ornithopodioides L. were evaluated in greenhouse and compared with Australian cultivar : Trifolium michelianum Savi. Paradana. Plants were submitted to three irrigation treatments using concentrations 0 mM, 80 mM and 200 mM of NaCl, during 3 months. The effects of salinity on growth, dry matter production, shoot and root ion relations, leaf water relation and gas exchange were determined. Under stressed conditions, T. isthmocarpum was more salt tolerant (p < 0.001) with a greater survival rate and good dry matter production, than L. ornithopodioides and T. michelianum . To tolerate salinity, T. isthmocarpum and T. michelianum developed a Cl - exclusion mechanism. However, the Cl - exclusion was more important in T. isthmocarpum than T. michelianum . On the other hand, T. isthmocarpum accumulated more K + in shoots than other species, when exposed to moderate and high levels of salinity. At 200 mM of NaCl, the stress promoted a substantial degree of stomatal regulation; but, in spite of this, L. ornithopodioides showed signs of leaf tissue dehydration, decreases in relative water content and osmotic potential values.
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Seasonal Dynamics of Stress Proteins in Leaves of Medicinal Plants in a Natural Environment of Irkutsk and on the Shores of the Lake Baikal
стр.42-50
Zhivetiev M.A., Kolesnichenko A.V., Graskova I.A., Voinikov V.K.
We study leafs of five plant species, growing in Irkutsk city and on the southeastern shore of Lake Baikal. These species are Achillea asiatica Serg., Taraxacum officinale Wigg., Plantago major L., Veronica chamaedrys L. and Alchemilla subcrenata Buser. In its leafs we identify some types of stress-induced proteins. In autumn, the accumulation of stress proteins in leafs of plants both from shores of Lake Baikal and from Irkutsk have been registered.
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Diurnal Influence on Phenol Compound Dynamic into Leaves of Medicinal Plants
стр.51-55
Zhivetiev M.A., Rudicovskaja E.G., Dudareva L.V., Graskova I.A., Voinikov V.K.
Content of phenol compound in leaves of Alchemilla subcrenata , Achillea asiatica , Taraxacum officinale , Veronica chamaedrys was researched. The dynamics of alteration of phenol compound content during round the clock in October was determined. The dependence of phenol compound substance in leaves of study plants on temperature was shown.
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Rhythmical changes of a level nitric oxide (NO) in roots etiolated seedlings of pea ( Pisum sativum L.) and influence of exogenous calcium
стр.56-66
Glyan’ko A.K., Ischenko A.A.
Studied time dynamics (during 60 mines) a level oxide nitric (NO) in cross cuts of roots 2 - day etiolated seedlings of pea sowing ( Pisum sativum L.) by use of fluorescent probe DAF-2DA and a fluorescent microscope depending on action exogenous calcium (Ca 2+). During an exposition of seedlings on water, solution CaCl 2 are shown fluctuation in level NO in roots - his increase and decrease that testifies to the certain rhythm in generation NO. Exogenous factors (Ca 2+) change time dynamics of level NO in comparison with variant “water”. Ca 2+chelate EGTA removes action exogenous calcium on rhythmical change of a level NO in roots. Results are discussed in aspect of close interference of signaling systems and molecules (Ca 2+, NO, Н 2О 2).
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The Role of Sugar-related Regulation in the Light-dependent Alterations of Arabidopsis Glutamate Dehydrogenase Genes Expression
стр.67-76
Garnik E.Yu., Belkov V.I., Tarasenko V.I., Konstantinov Yu.M.
Expression of gdh1 and gdh2 genes of Arabidopsis thaliana increases in the dark and decreases in the light. The reason of such alteration seems to be a glucose rising in photosynthetic cell in the light, but this hypothesis needs to be confirmed. In this work we investigate the role of glucose and hexokinase 1 in the light-dependent regulation of the gdh1 and gdh2 expression. A comparison of expression profiles of apl3 , gdh1 , gdh2 genes in presenсe of exogenous sucrose in the dark and in the light has demonstrated that sugar-related repression of gdh1 and gdh2 genes is insufficient to provide the high decrease of their transcripts in the light. Using Arabidopsis mutant gin2-1 with a defect in hxk1 gene we demonstrated that such a decrease is not depended on the regulatory function of hexokinase 1. We presume that light- dependent alterations of gdh1 and gdh2 expression are mediated by some chloroplast-to-nucleus regulatory signals.
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Studying of Import Mechanisms for Different Length and Structure DNA into Plant Mitochondria
стр.77-84
Klimenko E.S., Koulintchenko M.V., Grebnev P.A., Dietrich A., Konstantinov Yu.M.
We investigated import of DNA of various length (109 bp, 269 bp, 717 bp, 1540 bp, 2732 bp, 9000 bp and 11600 bp) into the isolated mitochondria from potato tubers ( Solanum tuberosum ) and turnip root crops ( Brassica rapa ). The results imply existence of multiple pathways of DNA transfer into plant mitochondria. It was established that the transport of short length DNA (100-300 bp) is apparently carried out with the participation of several protein carriers which nature is currently unknown. We identified factors that influence the process of DNA transfer into mitochondria. These factors include the length of nucleic acid molecule and the presence of terminal inverted repeats that are specific to linear mitochondrial plasmids of plants.
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Mitochondria as a Possible Place for Initial Stages of Steroid Biosynthesis in Plants
стр.85-97
Shematorova Elena K., Slovokhotov Ivan Yu., Khaliluev Marat R., Berdichevets Irina N., Baranova Ekaterina N., Babak Olga G., Shpakovski Dmitry G., Spivak Svetlana G., Shpakovski George V.
With the aim of thorough comparison of steroidogenic systems of plants and animals, transgenic plants of Solanaceae family expressing CYP11A1 cDNA encoding cytochrome P450 SCC of mammalian mitochondria were further analysed. Positive effect of CYP11A1 on resistance of the transgenic tobacco plants to the infection by fungal phytopathogene Botrytis cinerea was for the first time detected. Subtle changes in mitochondria of the transgenic Nicotiana tabacum plants expressing mammalian CYP11A1 cDNA were demonstrated by transmissive electron microscopy. Genetic determinants (cDNAs and genes) encoding main components of the electron transfer chain of plant mitochondria were for the first time cloned and characterized. It was established that plants from the Solanaceae family (tomato, tobacco and potato) contain two different genes with similar exon-intron structures (all contain 8 exons) encoding mitochondrial type ferredoxins (MFDX), and one gene for mitochondrial ferredoxin reductase (MFDXR). The results obtained point out on profound relatedness of electron transfer chains of P450-dependent monooxygenases in mammalian and plant mitochondria and support our previous findings about functional compatability of steroidogenic systems of Plantae and Animalia.
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Redox Enzymes of Red Beetroot Vacuoles ( Beta vulgaris L.)
стр.98-117
Pradedova E.V., Nimaeva O.D., Salyaev R.K.
Years of research have shown that some of the redox elements (enzymes, coenzymes, and co-substrate) are isolated from each other kinetic and spatial manner (compartmentalization) in the eukaryotic cells. The redox elements forming the "highly" and "widely" specialized redox system are found in all cell structures: mitochondria, plastids, peroxisomes, apoplast, nucleus etc. In recent years the active involvement of the central vacuole in the maintenance of the plant cell redox homeostasis is discussed, actually the information about the vacuolar redox system is very small. The high-priority redox processes and "redox-specialization" of the vacuolar compartment are not known. We have begun a study of red beet-root vacuole redox systems ( Beta vulgaris L.) and have identified redox enzymes such as: phenol peroxidase (EC 1.11.1.7), superoxide dismutase (EC 1.15.1.1) and glutathione reductase (EC 1.8.1.7). This paper presents some of the characteristics of these enzymes and considers the probable ways of their functioning in vacuolar redox chains.
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The Influence of Carbohydrate Status and Low Temperature on the Respiratory Metabolism of Mitochondria from Etiolated Leaves of Winter Wheat
стр.118-130
Borovik Olga A., Grabelnych Olga I., Koroleva Nina A., Pobezhimova Tamara P., Voinikov Victor K.
The separate and combined effect of sucrose (12%, 7 days) and low temperature (2 °С, 7 days) on the growth of plants, the content of carbohydrates in the leaves and oxidative activity of mitochondria isolated from them has been studied on the etiolated plants of winter wheat ( Triticum aestivum L.). It has been shown that sucrose and low temperature cause inhibition of the growth and increasing of the carbohydrates content. Using the different oxidation substrates (malate, malate + rotenone, succinate, NADH and NADPH) have been identified changes in the mitochondrial oxidative activity and the functioning of alternative oxidase and rotenone-insensitive NAD(P)H dehydrogenases. It has been determined that activity of the alternative oxidase and “external” rotenone-insensitive NAD(P)H dehydrogenases in the mitochondria of etiolated leaves depends on the carbohydrate status of the plant, regardless of the growth temperature.
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The Particularities of Nonspecific Stress Resistance Mechanism Activation in Baikalian Endemic Amphipod Gmelinoides fasciatus (Stebb, 1899) under Thermal Stress
стр.131-138
Vereshchagina K.P., Lubyaga Y.A., Axenov-Gribanov D.V., Gurkov A.N., Kondratieva E.S., Shchapova E.P., Prokosov S.O., Shatilina Z.M.
The aim of the current study was to determine the characteristics of the stress response in endemic species Gmelinoides fasciatus (Stebb., 1899) in comparison with other Baikal amphipods under short term thermal stress. It were shown the changes in activity of antioxidant enzymes, and lactate dehydrogenase correlated with content of lactate under the influence of gradual temperature increase. While the changes of cellular stress - markers content or their activity in other Baikal species occurred beyond the thermal preference zone, the cellular stress - markers content or their activity in G. fasciatus associated with entrance into the thermal preference zone. Thus we concluded that the relict endemic species G. fasciatus has a different stress resistance mechanism to elevated temperature than other Baikal amphipods species.
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The Impact of Experimental Hypoxia and Subsequent Normoxia on the Content of Some Ions and Markers of Physiological Stress-adaptation in Gastropod Species Lymnaea stagnalis
стр.139-146
Lubyaga J.А., Axenov-Gribanov D.V., Madyarova E.V., Gurkov А.N., Lozovoy D.V., Homich A.S., Golubev A.P., Timofeyev М.А.
The purpose of this study was to evaluate the influence of hypoxia and subsequent normoxiaon the maintenance of respiratory pigment hemocyanin, total protein, lactate and some ions (Na +, K +, Ca 2+, NH 4 +, Mg 2+) in the mantle liquid in palaearctic gastropod species Lymnaea stagnalis . It was shown that short-term experimental hypoxia leads to the activation of the physiological mechanisms of stress adaptation in widespread Palaearctic eurybiotic gastropod species and does not lead to the activation of the stress-resistance mechanisms on the biochemical and molecular levels.
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