Оценка антиоксидантного потенциала лекарственных растений при действии Уф-B-облучения

The UV-B irradiation action (6 -36,9 dg/m2) on medical plants Artemisia lerchiana L., Nigella sativa L., Ocimum basilicum L., grown in water culture, was studying. In plant leaves and roots the content of malondialdehyde, pigments, phenols, flavonoids, proline, and activities of antioxidant enzymes were determined. Investigated plants were able to resist to action of UV-B-irradiation (low and middle dose) without any power input for increasing of biosynthesis of low and high molecular component of antioxidant defense system. The damage level under stress conditions was low. At first hours plants used the constitutive level of antioxidants and after 24 h it was needed the antioxidant pool refilling. Carotinoids, flavonoids and proline were used by plants in 4 h after irradiation, the antocian synthesis increased after 12 h.В статье рассматривается методика изучения действия УФ-B-облучения (6,0, 12,3 и 36,9 Дж/кв. м) на лекарственные растения: полынь Artemisia lerchiana L., чернушку Nigella sativa L., базилик Ocimum basilicum L., выращенные в водной культуре в условиях фитотрона. Исследованные растения способны адаптироваться к действию низкой и средней доз УФ-B-радиации без значительных энергозатрат на усиление биосинтеза как низкомолекулярных компонентов антиоксидантной защиты, так и высокомолекулярных. Степень повреждения растений при действии данного стрессора незначительна. В первые часы растения в основном использовали конститутивный уровень антиоксидантов, и лишь через 24 часа требовалось пополнение их пула. Каротиноиды, флавоноиды и пролин используются растениями уже через 4 часа после облучения, синтез антоцианов усиливается через 12 ч

Участие антиоксидантной защитной системы базилика в кросс-адаптации при комплексном действии УФ-В и засоления

The 10 min UV-B irradiation (12 dj/m2) and 100mM NaCl action on medical plants Ocimum basilicum L., grown in water culture, was studying. The plants treated by UV-B irradiation were more resistent to salinity. There were two groups of compounds. One - proline, carotinoids, phenols are accumulated under salinity conditions. Second group - pigments - antocyans and flavones. Its biosynthesis is stimulated by UV-B irradiation. Under complex stressor action pigments were increased and this increasing helped plants to overcome the high salinity. The antioxidant enzyme superoxid dismutase did not play essential role in adaptive response. The low molecular weight antioxidants were more effective for formation of adaptation response. The peroxidase activity had not correlation with superoxid dismutase activity and we supposed that H2O2 could be result of other process and served as signal molecul.На основании полученных данных можно сделать главный вывод о том, что обработка УФ-В в течение 10 мин. способствовала адаптации растений базилика к действию 100 мМ NaCl. Улучшение адаптационной способности базилика при совместном действии УФ-В и засоления и связано со стимулированием биосинтеза пигментов и низкомолекулярных антиоксидантов УФ-В облучением

The involvement of low-molecular antioxidants in cross-adaptation of medicine plants to successive action of UV-B radiation and salinity

Artemisia (Artemisia lercheana Web.), common basil (Ocimum basilicum L.), and black cumin (Nigella sativa L.) plants grown in water culture until the stage of 4-5 true leaves were subjected to 10-min UV-B irradiation, treated with 100 mM NaCl, or subjected to the successive action of both stressors. The contents of proline, anthocyanins, flavonoids, soluble phenols, and carotenois were measured. Superoxide dismutase activity was also assayed. Experimental plants could tolerate UV-B irradiation due to the accumulation of phenolic compounds (anthocyanins, soluble phenols, and flavonoids). Anthocyanins contributed mostly in the defnse effect; their content in black cumin and common basil increased 3-5-fold after irradiation. Dynamics of the anthocyanin content in tested plants of all treatments indicates the activation of their biosynthesis by UV-B irradiation and suppression by salinity. Successive action of stressors resulted in synergism of their effects on accumulation of low-molecular compounds in artemisia and common cumin plants. When these plants were irradiated with UV-B and then treated with NaCl, they accumulated more low-molecular compounds than after separate treatment with these stressors, especially in Artemisia. Plant pretreatment with UV-B reduced the adverse action of salinity; this was manifested in the turgor maintenance in salt-treated plants after preliminary irradiation. For basil, salinity was the stronger stressor than UV-B, which was manifested in a decrease in the content of low-molecular compounds. © 2012 Pleiades Publishing, Ltd

Exogenous proline modifies differential expression of superoxide dismutase genes in UV-B-irradiated Salvia officinalis plants

Grown in water culture 6-week-old Salvia officinalis plants with 4-5 true leaves were exposed to irradiation with UV-B (10 min, 12. 3 kJ/m2), subjected to 5 mM exogenous proline in the nutrient solution, and treated with a combination of both factors. The plants responded to short UV-B irradiation by the appearance of oxidative stress, which was manifested in elevated content of malondialdehyde in leaves. Exogenous proline added 24 h before the irradiation inhibited lipid peroxidation. The total activity of superoxide dismutase (SOD) was analyzed in plant leaves, and three SOD isoforms-Mn-SOD, Fe-SOD, and Cu/Zn-SOD-were identified. Activities of these isoforms were measured over time, and the expression of their respective genes was analyzed by reverse transcription polymerase chain reaction (RT-PCR). It is shown that the addition of proline, UV-B irradiation, or combination of both treatments regulated in a differential manner the activities of SOD isoforms localized in various cell compartments. The activity of the cytosolic Cu/Zn-SOD isoform was limited by the presence of its mRNA, the content of which was regulated by mRNA synthesis or decay rate. By contrast, the activity of plastidic Fe-SOD isoform was regulated on the substrate (allosteric) level, not on the level of FSD gene expression. The activity of mitochondrial Mn-SOD isoform was insensitive to UV-B irradiation, addition of proline, or combination of both treatments, even though the level of MSD gene transcripts increased significantly after UV-B irradiation. The results indicate that MSD gene transcripts induced by UV-B were not completely processed to produce mature mRNA or mature mRNA was not capable of translation. It cannot be excluded that the synthesized macromolecule, the Mn-SOD precursor did not undergo posttranslational maturation to produce biologically active enzyme molecules. It appears that proline is involved in the differentially regulated complex expression of various SOD isoforms. This regulation is largely based on various extents of oxidative stress in different cell compartments. © 2011 Pleiades Publishing, Ltd

The involvement of low-molecular antioxidants in cross-adaptation of medicine plants to successive action of UV-B radiation and salinity

Artemisia (Artemisia lercheana Web.), common basil (Ocimum basilicum L.), and black cumin (Nigella sativa L.) plants grown in water culture until the stage of 4-5 true leaves were subjected to 10-min UV-B irradiation, treated with 100 mM NaCl, or subjected to the successive action of both stressors. The contents of proline, anthocyanins, flavonoids, soluble phenols, and carotenois were measured. Superoxide dismutase activity was also assayed. Experimental plants could tolerate UV-B irradiation due to the accumulation of phenolic compounds (anthocyanins, soluble phenols, and flavonoids). Anthocyanins contributed mostly in the defnse effect; their content in black cumin and common basil increased 3-5-fold after irradiation. Dynamics of the anthocyanin content in tested plants of all treatments indicates the activation of their biosynthesis by UV-B irradiation and suppression by salinity. Successive action of stressors resulted in synergism of their effects on accumulation of low-molecular compounds in artemisia and common cumin plants. When these plants were irradiated with UV-B and then treated with NaCl, they accumulated more low-molecular compounds than after separate treatment with these stressors, especially in Artemisia. Plant pretreatment with UV-B reduced the adverse action of salinity; this was manifested in the turgor maintenance in salt-treated plants after preliminary irradiation. For basil, salinity was the stronger stressor than UV-B, which was manifested in a decrease in the content of low-molecular compounds. © 2012 Pleiades Publishing, Ltd

Effect of common sage plant treatment with polyamines on the contents of proline and free and conjugated polyamines under oxidative stress

Common sage (Salvia officinalis L.) plants grown in water culture to the stage of 4-5 true leaves were treated with paraquat (PQ) (1 ml of the solution containing 0.1 μM PQ in 0.05% Tween 80), putrescine (Put), spermidine (Spd), or Spermine (Spm) (all polyamines (PA) at the concentration of 0.5 mM in 0. 05% Tween 80) or PA plus PQ for 6, 12, 24, 36, or 48 h. Under normal conditions, treatment with individual PA did not induce any changes in the content of free proline. Under oxidative stress induced by PQ, oxidation of proline and free PA by ROS resulted in their spending and demanded restoration of free PA due to hydrolysis of their insoluble conjugates. As distinct from treatment with PQ only, its combination with PA in the light was accompanied by proline accumulation. Treatment with Put plus PQ induced accumulation of intracellular Put and its soluble and insoluble conjugates. Treatments with Spd and Spm in combination with PQ resulted similarly in the increase in the levels of their intracellular soluble and insoluble conjugates. In treatments with these high-molecular PA, polyamine oxidase was activated and diaminopropan formation was observed. It seems likely that, for restoration of high-molecular PA homeostasis, their degradation by polyamine oxidase or production of insoluble conjugates is induced. Thus, the amount of free and conjugated particular PA is under strict control and is maintained at a definite level. A decrease in the content of a particular free PA induces primarily a decrease in the content of its soluble and then insoluble conjugates. The data obtained demonstrate the effects of exogenous PA on the content of intracellular proline under oxidative stress but do not allow a conclusion about direct regulation of its content by PA. © 2011 Pleiades Publishing, Ltd

Proline and functioning of the antioxidant system in Thellungiella salsuginea plants and cultured cells subjected to oxidative stress

The effects of proline on the functioning of antioxidant enzymes - superoxide dismutase (SOD) and ascorbate peroxidase (APO) - in Thellungiella salsuginea plants and cultured cells under normal conditions of culturing and under the influence of hydrogen peroxide (500 μM) were studied. Proline addition (0.2, 2, or 5 mM) to the medium for suspension culture or nutrient medium for plant growing resulted in the increase in the content of intracellular proline in both cultured cells and intact plant leaves and also in the activation of proline dehydrogenase, i. e., the enzyme degrading proline. Under normal conditions, treatment with proline exerted prooxidant action on both cellular and organismal levels. This was manifested in MDA accumulation and changes in APO and SOD activities. The amino acid alanine, used as a control, did not exert similar strong effect as proline. Application of 500 μM H2O2 on plant leaves resulted in the development of oxidative stress, whereas hydrogen peroxide addition into the culture medium - to the death of 50% of suspension cells. When plants and cultured cells were treated with 2 mM proline and than with H2O2, the number of dead cells in suspension was 35%, the content of MDA was decreased, APO was activated, and SOD activity was decreased in both cell culture and plant leaves. Thus, an increase in the intracellular proline concentration changed the redox balance and induced functioning of APO and SOD at both normal conditions of plant growing and cell culturing and under stress. © 2013 Pleiades Publishing, Ltd

Effect of common sage plant treatment with polyamines on the contents of proline and free and conjugated polyamines under oxidative stress

Common sage (Salvia officinalis L.) plants grown in water culture to the stage of 4-5 true leaves were treated with paraquat (PQ) (1 ml of the solution containing 0.1 μM PQ in 0.05% Tween 80), putrescine (Put), spermidine (Spd), or Spermine (Spm) (all polyamines (PA) at the concentration of 0.5 mM in 0. 05% Tween 80) or PA plus PQ for 6, 12, 24, 36, or 48 h. Under normal conditions, treatment with individual PA did not induce any changes in the content of free proline. Under oxidative stress induced by PQ, oxidation of proline and free PA by ROS resulted in their spending and demanded restoration of free PA due to hydrolysis of their insoluble conjugates. As distinct from treatment with PQ only, its combination with PA in the light was accompanied by proline accumulation. Treatment with Put plus PQ induced accumulation of intracellular Put and its soluble and insoluble conjugates. Treatments with Spd and Spm in combination with PQ resulted similarly in the increase in the levels of their intracellular soluble and insoluble conjugates. In treatments with these high-molecular PA, polyamine oxidase was activated and diaminopropan formation was observed. It seems likely that, for restoration of high-molecular PA homeostasis, their degradation by polyamine oxidase or production of insoluble conjugates is induced. Thus, the amount of free and conjugated particular PA is under strict control and is maintained at a definite level. A decrease in the content of a particular free PA induces primarily a decrease in the content of its soluble and then insoluble conjugates. The data obtained demonstrate the effects of exogenous PA on the content of intracellular proline under oxidative stress but do not allow a conclusion about direct regulation of its content by PA. © 2011 Pleiades Publishing, Ltd

Proline and functioning of the antioxidant system in Thellungiella salsuginea plants and cultured cells subjected to oxidative stress

The effects of proline on the functioning of antioxidant enzymes - superoxide dismutase (SOD) and ascorbate peroxidase (APO) - in Thellungiella salsuginea plants and cultured cells under normal conditions of culturing and under the influence of hydrogen peroxide (500 μM) were studied. Proline addition (0.2, 2, or 5 mM) to the medium for suspension culture or nutrient medium for plant growing resulted in the increase in the content of intracellular proline in both cultured cells and intact plant leaves and also in the activation of proline dehydrogenase, i. e., the enzyme degrading proline. Under normal conditions, treatment with proline exerted prooxidant action on both cellular and organismal levels. This was manifested in MDA accumulation and changes in APO and SOD activities. The amino acid alanine, used as a control, did not exert similar strong effect as proline. Application of 500 μM H2O2 on plant leaves resulted in the development of oxidative stress, whereas hydrogen peroxide addition into the culture medium - to the death of 50% of suspension cells. When plants and cultured cells were treated with 2 mM proline and than with H2O2, the number of dead cells in suspension was 35%, the content of MDA was decreased, APO was activated, and SOD activity was decreased in both cell culture and plant leaves. Thus, an increase in the intracellular proline concentration changed the redox balance and induced functioning of APO and SOD at both normal conditions of plant growing and cell culturing and under stress. © 2013 Pleiades Publishing, Ltd

Exogenous proline modifies differential expression of superoxide dismutase genes in UV-B-irradiated Salvia officinalis plants

Grown in water culture 6-week-old Salvia officinalis plants with 4-5 true leaves were exposed to irradiation with UV-B (10 min, 12. 3 kJ/m2), subjected to 5 mM exogenous proline in the nutrient solution, and treated with a combination of both factors. The plants responded to short UV-B irradiation by the appearance of oxidative stress, which was manifested in elevated content of malondialdehyde in leaves. Exogenous proline added 24 h before the irradiation inhibited lipid peroxidation. The total activity of superoxide dismutase (SOD) was analyzed in plant leaves, and three SOD isoforms-Mn-SOD, Fe-SOD, and Cu/Zn-SOD-were identified. Activities of these isoforms were measured over time, and the expression of their respective genes was analyzed by reverse transcription polymerase chain reaction (RT-PCR). It is shown that the addition of proline, UV-B irradiation, or combination of both treatments regulated in a differential manner the activities of SOD isoforms localized in various cell compartments. The activity of the cytosolic Cu/Zn-SOD isoform was limited by the presence of its mRNA, the content of which was regulated by mRNA synthesis or decay rate. By contrast, the activity of plastidic Fe-SOD isoform was regulated on the substrate (allosteric) level, not on the level of FSD gene expression. The activity of mitochondrial Mn-SOD isoform was insensitive to UV-B irradiation, addition of proline, or combination of both treatments, even though the level of MSD gene transcripts increased significantly after UV-B irradiation. The results indicate that MSD gene transcripts induced by UV-B were not completely processed to produce mature mRNA or mature mRNA was not capable of translation. It cannot be excluded that the synthesized macromolecule, the Mn-SOD precursor did not undergo posttranslational maturation to produce biologically active enzyme molecules. It appears that proline is involved in the differentially regulated complex expression of various SOD isoforms. This regulation is largely based on various extents of oxidative stress in different cell compartments. © 2011 Pleiades Publishing, Ltd