干旱胁迫下H_2S对板栗幼苗根系抗氧化特性及呼吸相关酶活性的影响
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摘要
为探究干旱胁迫下H2S对板栗幼苗根系特性的影响,以黄棚板栗幼苗为试验材料,通过PEG模拟干旱并进行H2S供体硫氢化钠(NaHS)和清除剂次牛磺酸(HT)处理,分析板栗幼苗根系超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)等抗氧化酶活性及丙二醛(MDA)、H2O2等抗氧化物质含量,检测苹果酸脱氢酶(MDH)、磷酸果糖激酶(PFK)和葡萄糖-6-磷酸脱氢酶(G-6-PDH)等呼吸代谢相关酶活性。结果表明,0. 5 mmol·L~(-1)NaHS+15%PEG协同处理24 h后,与15%PEG处理相比,根系SOD、POD、CAT、APX活性分别提高19. 64%、24. 67%、56. 91%、16. 29%,根系脯氨酸含量增加15. 25%,根系MDA和H2O2含量分别降低26. 52%和20. 86%,MDH、PFK和G-6-PDH活性分别提高27. 28%、23. 14%和15. 36%;而添加H2S清除剂HT则逆转了上述变化,说明适宜浓度的NaHS能够减轻干旱胁迫对板栗根系膜脂过氧化伤害和对呼吸代谢的抑制,从而有利于提高板栗幼苗对干旱逆境的适应能力。本研究结果为进一步探索H2S缓解板栗树干旱胁迫的机理提供了理论依据。
In order to investigate effect of H2 S on root characteristics of chestnut( Castanea mollissima) seedlings under drought stress,the chestnut seedlings of Huangpeng were used as experimental material,the antioxidant enzyme( SOD,POD,CAT,APX) activities,the content of MDA and H2 O2,and the respiration metabolism enzymes( MDH,PFK,G-6-PDH) activities were analyzed under drought stress induced by 15% PEG and treated with the H2 S donor sodium hydrosulfide( NaHS) and scavenger hypothiosulfate( HT) in this study. The results showed that under the treatment of0. 5 mmol·L~(-1) NaHS+15% PEG for 24 h,compared to 15% PEG treatment,the activity of SOD,POD,CAT and APX in root was increased by 19. 64%,24. 67%,56. 91% and 16. 29%,respectively; the proline content increased by15. 25%; the contents of MDA and H2 O2 were significantly decreased by 26. 52% and 20. 86%,respectively; and the activities of MDH,PFK and G-6-PDH were significantly increased by 27. 28%,23. 14%,15. 36%,respectively.However,the addition of H2 S scavenger HT reversed the above effects. The results showed that appropriate concentration of exogenous H2 S was favorable to reduce membrane lipid peroxidation damage and alleviate inhibition of respiratory metabolism in the root of chestnut seedlings under drought stress,thus improving the adaptability of chestnut seedlings to drought stress. This study provided a theoretical basis for further exploring the mechanism of H2 S alleviating drought stress in chestnut trees.
In order to investigate effect of H2 S on root characteristics of chestnut( Castanea mollissima) seedlings under drought stress,the chestnut seedlings of Huangpeng were used as experimental material,the antioxidant enzyme( SOD,POD,CAT,APX) activities,the content of MDA and H2 O2,and the respiration metabolism enzymes( MDH,PFK,G-6-PDH) activities were analyzed under drought stress induced by 15% PEG and treated with the H2 S donor sodium hydrosulfide( NaHS) and scavenger hypothiosulfate( HT) in this study. The results showed that under the treatment of0. 5 mmol·L~(-1) NaHS+15% PEG for 24 h,compared to 15% PEG treatment,the activity of SOD,POD,CAT and APX in root was increased by 19. 64%,24. 67%,56. 91% and 16. 29%,respectively; the proline content increased by15. 25%; the contents of MDA and H2 O2 were significantly decreased by 26. 52% and 20. 86%,respectively; and the activities of MDH,PFK and G-6-PDH were significantly increased by 27. 28%,23. 14%,15. 36%,respectively.However,the addition of H2 S scavenger HT reversed the above effects. The results showed that appropriate concentration of exogenous H2 S was favorable to reduce membrane lipid peroxidation damage and alleviate inhibition of respiratory metabolism in the root of chestnut seedlings under drought stress,thus improving the adaptability of chestnut seedlings to drought stress. This study provided a theoretical basis for further exploring the mechanism of H2 S alleviating drought stress in chestnut trees.
引文
[1]杨鹏辉,李贵全,郭丽,吴慎杰.干旱胁迫对不同抗旱大豆品种花荚期质膜透性的影响[J].干旱地区农业研究,2003,21(3):127-130
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[3] Guo W H,Li B,Huang Y M,Zhao H X,Zhang X S. Effects ofdifferent water stresses on eco-physiological characteristics ofHippophae rhamnoides seedlings[J]. Acta Botanica Sinica,2003,45(10):1238-1244
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[9]崔为体,沈文飚.植物中硫化氢的生理功能及其分子机理[J].生命的化学,2012,32(4):385-389
[10] Jin Z P,Pei Y X. Physiological implications of hydrogen sulfide inplant:pleasant exploration behind its unpleasant odour[J].Oxidative Medicine and Cellular Longevity,2015:397502-397507
[11]田雲,蒋景龙,李丽,余妙,任绪明.信号分子硫化氢调控植物抗逆性研究进展[J].核农学报,2017,31(11):2279-2287
[12] García-Mata C, Lamattina L. Hydrogen sulfide, a novelgasotransmitter involved in guard cell signaling[J]. New Phytologist,2010,188(4):977-984
[13] Zhang H,Ye Y K,Wang S H,Luo J P,Tang J,Ma D F. Hydrogensulfide counteracts chlorophyll loss in sweet potato seedling leavesand alleviates oxidative damage against osmotic stress[J]. PlantGrowth Regulation,2009,58(3):243-250
[14] Jin Z P,Shen J J,Qiao Z J,Yang G,Wang R,Pei Y X. Hydrogensulfide improves drought resistance in Arabidopsis thaliana[J].Biochemical and Biophysical Research Communications,2011,414(3):481-486
[15]刘晶,张鹤婷,殷悦,陈惠萍.外源硫化氢对干旱胁迫下萌发水稻种子抗氧化代谢的影响[J].南方农业学报,2017,48(1):31-37
[16]单长卷,赵元增.外源硫化氢对干旱胁迫下玉米幼苗水分生理特性的影响[J].干旱地区农业研究,2015,33(5):81-85
[17]张蜀秋,李云,武维华.植物生理学实验技术教程[M].北京:科学出版社,2011
[18]赵世杰,史国安,董新纯.植物生理学实验指导[M].北京:中国农业科学技术出版社,2002
[19]李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2003
[20] Miller G,Shulaev V,Mittler R. Reactive oxygen signaling andabiotic stress[J].Physiologia Plantarum,2008,133(3):481-489
[21]薛鑫,张芊,吴金霞.植物体内活性氧的研究及其在植物抗逆方面的应用[J].生物技术通报,2013(10):6-11
[22]谢志玉,张文辉,刘新成.干旱胁迫对文冠果幼苗的生长和生理生化特征的研究[J].西北植物学报,2010,30(5):948-954
[23]高相彬,赵凤霞,沈向,胡艳丽,郝云红,杨树泉,苏立涛,毛志泉.肉桂酸对平邑甜茶幼苗根系呼吸速率及相关酶活性的影响[J].中国农业科学,2009,42(12):4308-4314
[24] Nathan C,Ding A. Snapshot:reactive oxygen intermediates(ROI)[J]. Cell,2010,140(6):951
[25] Kornyeyev D,Logan B A,Payton P,Allen R D,Holaday A S.Enhanced photochemical light utilization and decreased chilling-induced photoinhibition of photosystemⅡin cotton overexpressinggenes encoding chloroplast-targeted antioxidant enzymes[J].Physiologia Plantarum,2001,113(3):323-331
[26]李玲,李俊,张春雷,张树杰,马霓,李光明.外源ABA和BR在提高油菜幼苗耐渍性中的作用[J].中国油料作物学报,2012,34(5):489-495
[27] Zhang H,Tan Z Q,Hu L Y,Wang S H,Luo J P,Jones R L.Hydrogen sulfide alleviates aluminum toxicity in germinating wheatseedlings[J]. Journal of Integrative Plant Biology,2010,52(6):556-567
[28]孟丹,刘玲,陈露,周思敏,安敏敏,罗玉明,杨立明.外源硫化氢对铝胁迫下水稻幼苗生长及生理生化的影响[J].江苏农业科学,2014,42(6):63-67
[29] Cui W,Chen H,Zhu K,Jin Q,Xie Y,Cui J,Xia Y,Zhang J,Shen W B. Cadmium-induced hydrogen sulfide synthesis is involvedin cadmium tolerance in Medicago sativa by reestablishment ofreduced(homo)glutathione and reactive oxygen specieshomeostases[J]. PLoS One,2014,9(10):e109669
[30] Zhang H,Hu L Y,Hu K D,He Y D,Wang S H,Luo J P.Hydrogen sulfide promotes wheat seed germination and alleviatesoxidative damage against copper stress[J]. Journal of IntegrativePlant Biology,2008,50(12):1518-1529
[31]孙晓莉,田寿乐,沈广宁,许林.外源Na HS对盐胁迫下板栗幼苗的缓解效应[J].山东农业科学,2017,49(4):51-54
[32] Wang Y Q,Li L,Cui W T,Xu S,Shen W B,Wang R. Hydrogensulfide enhances alfalfa(Medicago sativa)tolerance against salinityduring seed germination by nitric oxide pathway[J]. Plant and Soil,2012,351(1/2):107-119
[33]李永生,方永丰,李玥,慕平,王芳,张同祯,闫慧萍,王威,张金文,王汉宁.外源硫化氢对PEG模拟干旱胁迫下玉米种子萌发及幼苗生长的影响[J].核农学报,2016,30(4):813-821
[34] Zhang H,Jiao H,Jiang C X,Wang S,Wei Z J,Luo J P,JonesRL. Hydrogen sulfide protects soybean seedlings against drought-induced oxidative stress[J]. Acta Physiologiae Plantarum,2010,32(5):849-857
[35] Wu S,Hu C,Tan Q,Li L,Shi K,Zheng Y,Sun X. Drought stresstolerance mediated by zinc-induced antioxidative defense andosmotic adjustment in cotton(Gossypium Hirsutum)[J]. ActaPhysiologiae Plantarum,2015,37(8):167-175
[36]汤佩松.高等植物呼吸代谢途径的调节控制和代谢与生理功能间的相互制约[J].植物学报,1979,21(2):93-106
[37]李志霞,秦嗣军,吕德国,聂继云.植物根系呼吸代谢及影响根系呼吸的环境因子研究进展[J].植物生理学报,2011,47(10):957-966
[38]张鹏,秦嗣军,周文杰,吕德国,马怀宇.荧光假单胞菌对樱桃根系呼吸和幼苗生长的影响[J].中国农业科学,2014,47(19):3857-3865
[2] Costa J H,Jolivet Y,Hasenfratz-Sauder M P,Orellano E G,LimaaM G S,Dizengremel P,Fernandes D. Alternative oxidase regulationin roots of Vigna unguiculata cultivars differing in drought/salttolerance[J]. Plant Physiology,2007,164(6):718-727
[3] Guo W H,Li B,Huang Y M,Zhao H X,Zhang X S. Effects ofdifferent water stresses on eco-physiological characteristics ofHippophae rhamnoides seedlings[J]. Acta Botanica Sinica,2003,45(10):1238-1244
[4]田寿乐,沈广宁,许林,孙晓莉.不同节水灌溉方式对干旱山地板栗生长结实的影响[J].应用生态学报,2012,23(3):639-644
[5]孙晓莉,张鑫荣,田寿乐,沈广宁,许林.外源硫化氢处理对板栗幼苗干旱胁迫抗性的影响[J].北方园艺,2017(15):7-12
[6] Wang R. Physiological implications of hydrogen sulfide:a whiffexploration that blossomed[J]. Physiological Reviews,2012,92(2):791-896
[7]孙晓莉,姜倩倩,田寿乐,沈广宁,许林.新型气体信号分子H2S在植物中的研究进展[J].山东农业科学,2016,48(9):151-156,161
[8]金竹萍,裴雁曦.植物H2S气体信号分子的生理功能研究进展[J].中国细胞生物学学报,2013,35(6):880-886
[9]崔为体,沈文飚.植物中硫化氢的生理功能及其分子机理[J].生命的化学,2012,32(4):385-389
[10] Jin Z P,Pei Y X. Physiological implications of hydrogen sulfide inplant:pleasant exploration behind its unpleasant odour[J].Oxidative Medicine and Cellular Longevity,2015:397502-397507
[11]田雲,蒋景龙,李丽,余妙,任绪明.信号分子硫化氢调控植物抗逆性研究进展[J].核农学报,2017,31(11):2279-2287
[12] García-Mata C, Lamattina L. Hydrogen sulfide, a novelgasotransmitter involved in guard cell signaling[J]. New Phytologist,2010,188(4):977-984
[13] Zhang H,Ye Y K,Wang S H,Luo J P,Tang J,Ma D F. Hydrogensulfide counteracts chlorophyll loss in sweet potato seedling leavesand alleviates oxidative damage against osmotic stress[J]. PlantGrowth Regulation,2009,58(3):243-250
[14] Jin Z P,Shen J J,Qiao Z J,Yang G,Wang R,Pei Y X. Hydrogensulfide improves drought resistance in Arabidopsis thaliana[J].Biochemical and Biophysical Research Communications,2011,414(3):481-486
[15]刘晶,张鹤婷,殷悦,陈惠萍.外源硫化氢对干旱胁迫下萌发水稻种子抗氧化代谢的影响[J].南方农业学报,2017,48(1):31-37
[16]单长卷,赵元增.外源硫化氢对干旱胁迫下玉米幼苗水分生理特性的影响[J].干旱地区农业研究,2015,33(5):81-85
[17]张蜀秋,李云,武维华.植物生理学实验技术教程[M].北京:科学出版社,2011
[18]赵世杰,史国安,董新纯.植物生理学实验指导[M].北京:中国农业科学技术出版社,2002
[19]李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2003
[20] Miller G,Shulaev V,Mittler R. Reactive oxygen signaling andabiotic stress[J].Physiologia Plantarum,2008,133(3):481-489
[21]薛鑫,张芊,吴金霞.植物体内活性氧的研究及其在植物抗逆方面的应用[J].生物技术通报,2013(10):6-11
[22]谢志玉,张文辉,刘新成.干旱胁迫对文冠果幼苗的生长和生理生化特征的研究[J].西北植物学报,2010,30(5):948-954
[23]高相彬,赵凤霞,沈向,胡艳丽,郝云红,杨树泉,苏立涛,毛志泉.肉桂酸对平邑甜茶幼苗根系呼吸速率及相关酶活性的影响[J].中国农业科学,2009,42(12):4308-4314
[24] Nathan C,Ding A. Snapshot:reactive oxygen intermediates(ROI)[J]. Cell,2010,140(6):951
[25] Kornyeyev D,Logan B A,Payton P,Allen R D,Holaday A S.Enhanced photochemical light utilization and decreased chilling-induced photoinhibition of photosystemⅡin cotton overexpressinggenes encoding chloroplast-targeted antioxidant enzymes[J].Physiologia Plantarum,2001,113(3):323-331
[26]李玲,李俊,张春雷,张树杰,马霓,李光明.外源ABA和BR在提高油菜幼苗耐渍性中的作用[J].中国油料作物学报,2012,34(5):489-495
[27] Zhang H,Tan Z Q,Hu L Y,Wang S H,Luo J P,Jones R L.Hydrogen sulfide alleviates aluminum toxicity in germinating wheatseedlings[J]. Journal of Integrative Plant Biology,2010,52(6):556-567
[28]孟丹,刘玲,陈露,周思敏,安敏敏,罗玉明,杨立明.外源硫化氢对铝胁迫下水稻幼苗生长及生理生化的影响[J].江苏农业科学,2014,42(6):63-67
[29] Cui W,Chen H,Zhu K,Jin Q,Xie Y,Cui J,Xia Y,Zhang J,Shen W B. Cadmium-induced hydrogen sulfide synthesis is involvedin cadmium tolerance in Medicago sativa by reestablishment ofreduced(homo)glutathione and reactive oxygen specieshomeostases[J]. PLoS One,2014,9(10):e109669
[30] Zhang H,Hu L Y,Hu K D,He Y D,Wang S H,Luo J P.Hydrogen sulfide promotes wheat seed germination and alleviatesoxidative damage against copper stress[J]. Journal of IntegrativePlant Biology,2008,50(12):1518-1529
[31]孙晓莉,田寿乐,沈广宁,许林.外源Na HS对盐胁迫下板栗幼苗的缓解效应[J].山东农业科学,2017,49(4):51-54
[32] Wang Y Q,Li L,Cui W T,Xu S,Shen W B,Wang R. Hydrogensulfide enhances alfalfa(Medicago sativa)tolerance against salinityduring seed germination by nitric oxide pathway[J]. Plant and Soil,2012,351(1/2):107-119
[33]李永生,方永丰,李玥,慕平,王芳,张同祯,闫慧萍,王威,张金文,王汉宁.外源硫化氢对PEG模拟干旱胁迫下玉米种子萌发及幼苗生长的影响[J].核农学报,2016,30(4):813-821
[34] Zhang H,Jiao H,Jiang C X,Wang S,Wei Z J,Luo J P,JonesRL. Hydrogen sulfide protects soybean seedlings against drought-induced oxidative stress[J]. Acta Physiologiae Plantarum,2010,32(5):849-857
[35] Wu S,Hu C,Tan Q,Li L,Shi K,Zheng Y,Sun X. Drought stresstolerance mediated by zinc-induced antioxidative defense andosmotic adjustment in cotton(Gossypium Hirsutum)[J]. ActaPhysiologiae Plantarum,2015,37(8):167-175
[36]汤佩松.高等植物呼吸代谢途径的调节控制和代谢与生理功能间的相互制约[J].植物学报,1979,21(2):93-106
[37]李志霞,秦嗣军,吕德国,聂继云.植物根系呼吸代谢及影响根系呼吸的环境因子研究进展[J].植物生理学报,2011,47(10):957-966
[38]张鹏,秦嗣军,周文杰,吕德国,马怀宇.荧光假单胞菌对樱桃根系呼吸和幼苗生长的影响[J].中国农业科学,2014,47(19):3857-3865
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