Comparative Analysis of Some Hsps Synthesis in Leaves of Transgenic Potato Plants with Gene gox at High Temperature
Borovskii G.B., Lyubushkina I.V., Borovik O.A., Grabelnych O.I., Sauchyn D.V., Urbanovich O.Yu.
Analysis of control and transgenic plants of potato, grade Skarb, transformed by vector constructions pBI-L-GOX ( LP line) и pBI-F-GOX ( F line) with native gene of glucose oxidase gox and pBI-GOX-mod ( MOD line) with modified gene gox-mod has been carried out. It has been shown that transgenic potato plants with genes gox and gox-mod did not have fenotypic differences with control plants. It has been confirmed that there was an increase of level of hydrogen peroxide produced by reaction, which catalized by glucose oxidase in leaves of transgenic plants. Using antibodies against some heat shock proteins (Hsp101, Hsp70/Hsc70, Hsp60, Hsp17.6C-1, Hsp17.6C-2) we revealed an increased content of Hsp17.6C-2 in leaves of LP line and Hsp60 in leaves of LP and F in the morning. It has been shown that rise in temperature in the daytime (from 18-23°С to 35-43°С) led to intensification of all studied Hsps synthesis both in control and in transformed lines.
Salt Tolerance Evaluation in Cotton ( Gossypium hirsutum L.) Using RAPD Marker
The aim of this study was to evaluate four upland cotton ( Gossypium hirsutum L.) [Niab78 (N78), Deir-Ezzor22 (DE22), Deltapine50 (DP50) and Aleppo118 (A118)] varieties towards salt stress (0 and 200 mM NaCl) for 7 weeks based on RAPD marker. Our data showed that the highest total polymorphic bands identified by the 26 RAPD tested primers were 150 bands generated by N78 variety, while the lowest ones were recorded for DP50 (29 bands). Otherwise, unique (negative and positive) markers characterized the two tolerant varieties (N78 and DE22) were 22 and 29 markers, respectively. Our data indicated that the highest polymorphism level was detected in N78 variety (68.5%) followed by DE22 (60.9%), whereas, the lowest one was recorded for DP50 (21.3%). Our data obtained herein indicates that RAPD marker provided molecular markers for salinity tolerance screening in early stage in genetic improvement programs.
Comparative Evaluation of Physiological and Biochemical Parameters and Survival of Chickens for the Effects of Microclimatic Stress Factors During Incubation
The article presents the results of two experiments, the findings reflect the influence of temperature and moisture stress on physiological and biochemical parameters of chickens and their viability during embryonic and early postembryonic development. It was found that under the influence of stress factors recorded increase in lipid peroxidation in chickens from day-old experimental groups; showed a reduction in the intensity of the major metabolism of young chickens day old.
Initial Morphogenetic Features of Proteome of Suprastructures of Interphase Chromatin for Germination of Mature Germs in Conditions of Adapting to Winter in Wheat
Ivanova E.A., Vafina G.H., Ivanov R.S.
Investigation morphogenetic processes of how the plant is formed for interact with the environment is necessary to understanding the fundamental problems of stress supramolecular biochemistry of developmental biology. Modern biology has established that the molecular morphogenesis belonging to the number of morphological processes. The molecular morphogenesis is directly based on well-studied forces of intermolecular interactions. From the progress of understanding the morphogenesis, expected not only purely cognitive, but also the practical results of biotechnology. The purpose of work was the analysis of shielding of morphogenetic suprastructures of interphase chromatin matrix to the localization of sensitivity of the regulatory proteolysis in the conditions of adaptation of vegetative phase of wheat to the stressful environmental factors. It was shown that in the conditions of long-term adaptation to cold shock of wheat at the level of suprastructures, of the packaged in the nuclear matrix of the genome, is carried out the total shielding of arginine-X zones for the regulatory protease-processing. Perhaps these zones represents one of the elements of the LCR (locus control regions), which are related to the histone code and influence the architectural organization of the cell nucleus, which is formed to facilitate life of organism in complex environmental conditions. These data will be useful for those who involved in the development of mathematical-logic schemes of theory and practice of biological specificity and may be included in the ontology of stages of plant growth and development.
Phenotypic Traits of Arabidopsis Plants Deficient in RpoTmp RNA Polymerase
Tarasenko V.I., Garnik E.Yu., Koulintchenko M.V., Konstantinov Yu.M.
In Arabidopsis, three nuclear-encoded RNA polymerases participate in the transcription of organellar genes. RpoTmp is a RNA polymerase that localizes both in mitochondria and chloroplasts, but is involved predominantly in the control of gene expression in mitochondria. Insertion mutant rpotmp is characterized by a number of phenotypic and molecular-biological peculiarities including decreased activities of the mitochondrial respiratory complexes I and IV. In the present study we compared growth characteristics of the rpotmp mutant and fro1 mutant which is characterized by the absence of functional complex I. We showed that in spite of the similar molecular defects and phenotypic appearance, the investigated mutants can be distinguished by the growth rate under different photoperiod as well as by the age of leaf senescence onset. Moreover, the growth rate of suspension cell culture of the rpotmp line is extremely retarded which clearly distinguished it from the fro1 suspension cell culture. We propose that unique properties of the rpotmp mutant are associated with the decreased level of respiratory complex IV activity.
Activity of the Respiratory Chain Enzymes of Blood Leucocytes’ Mitochondria Under the Conditions of Toxic Hepatitis Induced Against the Background Alimentary Deprivation of Protein
Voloshchuk O.N., Kopylchuk G.P.
Full functioning of the leucocytes’ energy supply system is one of the essential factors for the immune surveillance system effective work. The pivotal enzymes of the leucocytes’ energy biotransformation system are NADH-ubiquitin reductase, a marker of the Complex I of respiratory chain activity, and succinate dehydrogenase, key enzyme of the Complex II of respiratory chain. The aim of research - to study the NADH-ubiquitin reductase and succinate dehydrogenase activity of the blood leucocytes’ mitochondria under the conditions of toxic hepatitis induced against the background alimentary deprivation of protein. It is shown, that under the conditions of acetaminophen-induced hepatitis a reduction of the NADH-ubiquitin reductase enzymatic activity is observed on the background activation of the succinate-dependent way of the mitochondrial oxidation. Conclusion was made that alimentary deprivation or protein is a factor, aggravating the misbalance of the energy biotransformation system in the leucocytes of rats with toxic hepatitis. Established activity changes of the leucocytes’ mitochondria respiratory chain key enzymes may be considered as one of the mechanisms, directed on the maintenance of leucocytes energy supply on a level, sufficient for their functioning. Research results may be used for the biochemical rationale of the therapeutic approaches to the elimination and correction of the leucocytes’ energy metabolism disturbances consequences under the conditions of acetaminophen-induced hepatitis, aggravated by the alimentary protein deprivation.
Effects of Light Intensity on Development and Chlorophyll Content in the Arabidopsis Mutant Plants with Defects in Photosynthesis
Garnik E.Yu., Deeva D.V., Belkov V.I., Tarasenko V.I., Konstantinov Yu.M.
The developmental stages and adaptability to different light intensity (150 µmol*m -2*s -1 and 100 µmol*m -2*s -1) in Arabidopsis mutant lines with defects of photosynthetic apparatus were analyzed. Plant development in the mutant lines depended on the light intensity to varying degrees. Lines ch1-1 (lack of the chlorophyllide a oxygenase) and rtn16 (decreased chlorophyll a and b amounts) were the most susceptible to the light decrease. No one of the investigated lines demonstrated chlorophyll a/b rate alteration under the different light conditions. The depleted chlorophyll content has had the major effect on the mutant plants development under the different light conditions. The different chlorophyll a/b rate correlated with the different adaptability of mutant plants to low light.
Implications of Plants Genetic Transformation Assessed by Geneticist, Biochemist and Physiologist
Enikeev A.G., Kopytina T.V., Maximova L.A., Nurminskaya J.V., Shvetsov S.G.
Transgenic plants creation methodology developed for several decades has gained significant advances. However, problems of unanticipated effects of transgenosis, stability of GMO characteristics and establishing criteria of their safety evaluation remain unresolved. The analysis of different approaches to assessing the impact of plant genetic transformation is presented. It is concluded that the profound studies on the physiology of plant-agrobacterial symbiosis as a methodological basis of plants genetic engineering can answer many unresolved issues of genetic engineering.
Absorption of Power Plants СО 2 Emissions by Coniferous Tree Stands
The article reviews the ability of coniferous (common pine, siberian larch and siberian spruce) stands growing in 9 municipal districts of the Irkutsk region to absorb СО 2 technogenic emission of heat power plants. (EIGAF) index is suggested to characterize gas-absorbing (СО 2-absorbing) activity; the index reflects proportion between СО 2 technogenic emission and photosynthetic productivity (GPP) of coniferous tree stands. СО 2-absorbing capacity in 8 of the monitored districts has been shown to significantly exceed the amount of carbon dioxide emission from heat power sector. The index values EIGAF=0.01-0.97 demonstrate that СО 2 technogenic emission amounts to 1-97% of coniferous stands photosynthetic productivity in the areas under study. At the same time, the most industrially developed Angarsk district shows СО 2 photosynthetic absorption to be 8-12 times lower than technogenic СО 2 emission. Reasons of low gas-absorbing capacity of coniferous tree stands of this area are discussed.
Effect of Temperature on Triticum aestivum L. Seedlings Growth and Phytohormone Balance
Kosakivska I.V., Voytenko L.V., Likhnyovskiy R.V.
He study aimed to determine the effect of short-term heat and cold stresses on growth, accumulation and distribution of abscisic acid (ABA) and indole-3-acetic acid (IAA) in 7 and 14-day-old Triticum aestivum L. seedlings of the frost resistant cultivar Volodarka. A high performance liquid chromatography method has been used to provide a qualitative and quantitative analysis of hormones in leaves and roots. Biometric analysis showed that at the early stage of vegetation (7 days) seedlings were more resistant to temperature stresses, especially to cold, than at the later ones (14 and 21 days), that correlates with the frost resistance of cultivar. The results indicated that at the early stages of growth, after a short-term cold stress (2 h., +2 0C), the amount of free ABA substantially increased in roots, while at the later ones, it occurred after a short-term heat stress (2 h., +40 0C) in leaves. After a heat stress the leaves of 7-day-old seedlings accumulated a conjugated form of IAA. Heat stress caused the accumulation of free IAA in roots of 14-day-old seedlings. Cross stress (cold+heat) caused some increase in the pool of endogenous ABA both in roots and leaves while the amount of free IAA increased only in leaves. Сhanges in the accumulation of the free and conjugated forms of ABA and IAA depended both on the organs and the age of seedlings, and the type of stress and correlated with frost resistance of the cultivar Volodarka.
The Effect of Herbicides on Hydrogen Peroxide Generation in Isolated Vacuoles of Red Beet Root ( Beta vulgaris L.)
Pradedova E.V., Nimaeva O.D., Karpova A.B., Salyaev R.K.
Influence of herbicides on the hydrogen peroxide generation in vacuolar extracts of red beet root ( Beta vulgaris L.) was investigated. Belonging to different chemical classes of herbicide compounds have been used. Herbicides differ from each other in the mechanism of effects on plants. Clopyralid (aromatic acid herbicide, derivative of picolinic acid) and 2.4-D (phenoxyacetic herbicide), characterized by hormone-like effects, contributed to the formation of H 2O 2 in vacuolar extracts. Fluorodifen (nitrophenyl ether herbicide) and diuron (urea herbicide) also have increased contents H 2O 2. These compounds inhibit the electron transport, photosynthesis, and photorespiration in sensitive plants. Herbicidal effect of glyphosate (organophosphorus herbicide) is due to the inhibition of amino acid synthesis in plant cells. Glyphosate did not affect the content of H 2O 2 in vacuolar extracts. Herbicide dependent H 2O 2-generation did not occur with oxidoreductase inhibitors, potassium cyanide and sodium azide. The results suggest that the formation of ROS in the vacuoles due to activity of oxidoreductases, which could interact with herbicides.
The Daily Dynamics of Phenolic Compounds Content and Guaiacol-Peroksidase Activity in Leaves and Flowers of Alchemilla subcrenata and Veronica chamaedrys
Zhivetev M.A., Dudareva L.V., Rudikovskaya E.G., Graskova I.A., Voinikov V.K.
The daily dynamics of peroxidase activity and the total content of phenolic compounds in leaves and inflorescences of Alchemilla subcrenata Buser and Veronica chamaedrys L. were studied. Significant changes in the content of phenolic compounds and activity peroksidase depending on the time of day was shown.
The Tebuconazole-based Protectant of Seeds “Bunker” Induces the Synthesis of Dehydrins During Cold Hardening and Increases the Frost Resistance of Wheat Seedlings
Korsukova A.V., Borovik O.A., Grabelnych O.I., Voinikov V.K.
Triazole derivatives are widely used in agriculture for seed protectant of cereals against seed and soil infection. Triazole derivatives can have an effect on the biochemical and physiological functions of plants. The tebuconazole-based protectant of seeds «Bunker» (content of tebuconazole 60 grams per liter, g/L) is a systemic fungicide of preventive and therapeutic action. The effect of the seed treatment by «Bunker» preparation on the shoot growth and cell viability coleoptile, synthesis of dehydrins in shoots and frost resistance etiolated winter and spring wheat seedlings has been studied. It has been shown that treatment of winter and spring wheat seed by «Bunker» preparation induces similar concentration-dependent inhibition of the coleoptiles length. At the recommended dose (0,5 liter per tonne of seeds, L/t) growth inhibition was 28 - 30%, at a concentration of 1 L/t - 33 - 36%, at a concentration of 1,5 L/t - 40 - 42%, at a concentration of 3 L/t - 43 - 47%, at a concentration of 4 L/t - 48 - 51% and at 5 L/t - 53 - 56%. The treatment of wheat seed by «Bunker» preparation had no phytotoxic effect on coleoptile cells in any of the studied concentrations, on the contrary, with increasing concentration of preparation observed the increase in cell viability, as measured by recovery of 2,3,5-triphenyltetrazolium chloride. We can assume that having retardant properties, tebuconazole not only inhibits the growth of plants, but also delays their aging. The treatment of seed protectant at a concentration of 1.5 L/t induced synthesis of the dehydrins with molecular masses about 19, 21, 22, 25 and 27 kD in winter wheat shoots and 18,6, 27 and 28,5 kD in spring wheat shoots during cold hardening. Among identified dehydrins the dehydrin of 27 kD is most significantly induced both in winter and spring wheat. The treatment of seed protectant «Bunker» in the same concentration increased the frost resistance of winter and spring wheat seedlings. It is supposed that increasing of cold and frost resistance of winter and spring wheat caused by seed treatment of the tebuconazole-based protectant may be associated with increase of the abscisic acid content - one of the triggers of the low-temperature adaptation of plants. It is concluded that tebuconazole-based protectant of seeds «Bunker» increases the resistance of wheat to low temperature, affecting the growth processes and the synthesis of stress proteins.
Changing of the HSP70 Content in the Baikal Endemic Sponges Lubomirskiidae Under Conditions of Hyperthermia
Itskovich V.B., Shigarova A.M., Glyzina O.Y., Kaluzhnaya O.V., Kupchinsky A.B., Borovskii G.B.
Baikal endemic sponges (Lubomirskiidae) make up the bulk of the benthos biomass of the lake. For the first time the changes in the content of HSP70 in response to elevated environment temperature were analyzed in three endemic species of Baikal sponges: Baikalospongia bacillifera (Dybowski, 1880), B. intermedia (Dybowski, 1880) and Swartschewskia papyracea (Dybowski, 1880). Interspecific variability of constitutive HSP70 level was revealed for representatives of the three analyzed Lubomirskiidae species. After exposure at 13 °С for 3 and 7 days opposite changes were noted in the amount of HSP70. Under conditions of hyperthermia the protein level decrease at Baikalospongia species, while at the S. papyracea HSP70 content slightly increased. The differences in the mechanisms of stress adaptation probably affect the thermal resistance of the species, as well as are evidence supporting their specific status.
Salinity Stress Effects on Growth and Nitrate Assimilation in Bean Seedlings Likely to be Mediated via Nitric Oxide
Dhamgaye S., Gadre R.
Background : Salinity stress usually imposes adverse effects on plant systems, but the severity depends upon plant species, growth status and genotype, nutritional and environmental conditions etc. Present study analyses salinity effects on growth and in vivo nitrate reductase activity (NRA) in Phaseolus vulgaris seedlings to work out the mechanism. Results : Supply of 10-200 mM NaCl with 10 mM KNO 3 for 24 h in continuous light reduced the overall growth of the bean seedlings, with perfect -ve correlation for seedling weight, root length and shoot length. Salinity effect with 10 mM NH 4Cl was lesser and with 10 mM NH 4NO 3 was intermediary. NaCl treatment with 10 mM KNO 3 reduced the fresh wt of the root as well as shoot tissue but increased in vivo NRA exerting strong correlation and more prominent effect in the root tissue. Very high NaCl concentration prominently increased NRA at 1, 10 and 50 mM KNO 3 showing inverse gradation in effect. Salt treatment with NH 4NO 3, reduced the in vivo NRA of the root tissue, but the stress parameters, like proline and peroxidase were increased. Conclusions : The salinity stress effects on NRA are less severe with NH 4 +, more prominent for root and more effective at low NO 3 - concentration. Inverse correlation between decrease in fresh mass and increase in NRA with salinity suggests the involvement of NR in the synthesis of nitric oxide and the observed effect of stress is the balance of two factors reduced assimilatory effect and increased nitric oxide stress.