外源NO对Ca(NO_3)_2胁迫下番茄叶片活性氧损伤的缓解效应
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摘要
为探讨外源一氧化氮(NO)对次生盐渍胁迫下植物抗氧化系统的调节作用,以番茄品种‘秦丰保冠’为试材,在营养液栽培条件下研究叶面喷施外源NO供体硝普钠(SNP)对80mmol·L~(-1)Ca(NO_3)_2胁迫下番茄幼苗生长、叶片光合、活性氧物质、抗氧化酶活性和抗坏血酸-谷胱甘肽(AsA-GSH)循环的影响。结果表明,Ca(NO_3)_2胁迫下喷施SNP处理的番茄幼苗叶片超氧阴离子(O_2~(·-))的产生速率以及过氧化氢(H_2O_2)、丙二醛(MDA)、脱氢抗坏血酸(DHA)、氧化型谷胱甘肽(GSSG)的含量和电解质渗漏率显著降低,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)、谷胱甘肽还原酶(GR)、脱氢抗坏血酸还原酶(DHAR)和单脱氢抗坏血酸还原酶(MDHAR)的活性显著升高或得以维持,同时叶片抗坏血酸(AsA)、谷胱甘肽(GSH)含量及其还原力(AsA/DHA、GSH/GSSG值)显著升高,叶片活性氧损伤得到有效缓解,叶绿素降解和光合速率的下降得到有效抑制,进而促进了植株的生长发育,提高了番茄幼苗的耐盐能力。
【Objective】In relatively enclosed greenhouses, soil salts tend to accumulate gradually in topsoil duo to the special greenhouse environment of no rain water leaching, high temperature and resultant high evaporation, thus leading to serious secondary salinization of the greenhouse soil. According to relevant reports, the high level of Ca(NO_3)_2 accumulation was one of the main causes of high soil salinity in greenhouses. Nitric oxide(NO) is a kind of micromolecule active material that generally exists inside plants.As a plant hormone and signaling molecule, NO extensively participates in regulating of plants' responses to various adversity stress. The aim of this paper is to investigate roles of exogenous NO in regulating the antioxidant system of plants under Ca(NO_3)_2 stress. 【Method】A hydroponic experiment was conducted to investigate effects of foliar spray of sodium nitroprusside(SNP) as exogenous NO donor on growth of tomato(‘Qin Feng Bao Guan') seedlings, and photosynthetic, reactive oxygen, anti-oxidase activities,and ascorbate-glutathione cycle in their leaves under the stress of 80 mmol·L~(-1) Ca(NO_3)_2. 【Result】In the leaves of the tomato seedlings under Ca(NO_3)_2 stress, foliar spray SNP significantly lowered O_2~(·-) production rate, H_2O_2, malondialdehyde, dehydroascorbic acid and oxidized glutathione in content and electrolyte leakage rate, but heightened or maintained superoxide dismutase, peroxidase, catalase, ascorbate peroxidase,glutathione reductase, dehydroascorbate reductase and monodehydroascorbate reductase in activity; besides,it significantly increased the contents of ascorbic acid and glutathione and their reducing power. As a result, active oxygen damage of the leaves was effectively alleviated, and chlorophyll degradation and photosynthetic rate declining trend were effectively suppressed. 【Conclusion】Exogenous NO treatment plays an important role in maintaining or improving the activity of antioxidant enzymes and promoting the operation of aseorbate-glutathione(AsA-GSH) cycle in tomato under the stress of Ca(NO_3)_2, thus relieving the plants of oxidative damage significantly, and improving their leaf membrane system in stability, their photosynthetic function, their tolerance to salt and eventually their growth.
【Objective】In relatively enclosed greenhouses, soil salts tend to accumulate gradually in topsoil duo to the special greenhouse environment of no rain water leaching, high temperature and resultant high evaporation, thus leading to serious secondary salinization of the greenhouse soil. According to relevant reports, the high level of Ca(NO_3)_2 accumulation was one of the main causes of high soil salinity in greenhouses. Nitric oxide(NO) is a kind of micromolecule active material that generally exists inside plants.As a plant hormone and signaling molecule, NO extensively participates in regulating of plants' responses to various adversity stress. The aim of this paper is to investigate roles of exogenous NO in regulating the antioxidant system of plants under Ca(NO_3)_2 stress. 【Method】A hydroponic experiment was conducted to investigate effects of foliar spray of sodium nitroprusside(SNP) as exogenous NO donor on growth of tomato(‘Qin Feng Bao Guan') seedlings, and photosynthetic, reactive oxygen, anti-oxidase activities,and ascorbate-glutathione cycle in their leaves under the stress of 80 mmol·L~(-1) Ca(NO_3)_2. 【Result】In the leaves of the tomato seedlings under Ca(NO_3)_2 stress, foliar spray SNP significantly lowered O_2~(·-) production rate, H_2O_2, malondialdehyde, dehydroascorbic acid and oxidized glutathione in content and electrolyte leakage rate, but heightened or maintained superoxide dismutase, peroxidase, catalase, ascorbate peroxidase,glutathione reductase, dehydroascorbate reductase and monodehydroascorbate reductase in activity; besides,it significantly increased the contents of ascorbic acid and glutathione and their reducing power. As a result, active oxygen damage of the leaves was effectively alleviated, and chlorophyll degradation and photosynthetic rate declining trend were effectively suppressed. 【Conclusion】Exogenous NO treatment plays an important role in maintaining or improving the activity of antioxidant enzymes and promoting the operation of aseorbate-glutathione(AsA-GSH) cycle in tomato under the stress of Ca(NO_3)_2, thus relieving the plants of oxidative damage significantly, and improving their leaf membrane system in stability, their photosynthetic function, their tolerance to salt and eventually their growth.
引文
[1]Yuan L,Shu S,Sun J,et al.Effects of 24-epibrassinolide on the photosynthetic characteristics,antioxidant system,and chloroplast ultrastructure in Cucumis sativus L.under Ca(NO3)2 stress.Photosynthesis Research,2012,112(3):205-214
[2]孙德智,韩晓日,彭靖,等.外源NO对Ca(NO3)2胁迫下番茄幼苗PSII功能及光能分配利用的影响.核农学报,2016,30(12):2451-2459Sun D Z,Han X R,Peng J,et al.Effect of exogenous nitric oxide on PSII function and distribution and utilization of luminous energy in tomato seedlings under stress of Ca(NO3)2(In Chinese).Journal of Nuclear Agricultural Sciences,2016,30(12):2451-2459
[3]Delian E,Bǎdulescu L,Dobrescu A,et al.A brief overview of seed priming benefits in tomato.Romanian Biotechnological Letters,2017,22(3):12505-12513
[4]Santolini J,AndréF,Jeandroz S,et al.Nitric oxide synthase in plants:Where do we stand?Nitric Oxide,2017,63:30-38
[5]Hasanuzzaman M,Nahar K,Alam M M,et al.Exogenous nitric oxide alleviates high temperature induced oxidative stress in wheat(Triticum aestivum L.)seedlings by modulating the antioxidant defense and glyoxalase system.Australian Journal of Crop Science,2012,6(8):1314-1323
[6]Liang L,Deng Y,Sun X,et al.Exogenous nitric oxide pretreatment enhances chilling tolerance of anthurium.Journal of the American Society for Horticultural Science,2018,143(1):3-13
[7]Shan C,Zhou Y,Liu M.Nitric oxide participates in the regulation of the ascorbate-glutathione cycle by exogenous jasmonic acid in the leaves of wheat seedlings under drought stress.Protoplasma,2015,252(5):1397-1405
[8]Esringu A,Aksakal O,Tabay D,et al.Effects of sodium nitroprusside(SNP)pretreatment on UV-B stress tolerance in lettuce(Lactuca sativa L.)seedlings.Environmental Science and Pollution Research,2016,23(1):589-597
[9]Ahmad P,Ahanger M A,Alyemeni M N,et al.Exogenous application of nitric oxide modulates osmolyte metabolism,antioxidants,enzymes of ascorbate-glutathione cycle and promotes growth under cadmium stress in tomato.Protoplasma,2018,255(1):79-93
[10]Ahmad P,Abdel Latef A A,Hashem A,et al.Nitric oxide mitigates salt stress by regulating levels of osmolytes and antioxidant enzymes in chickpea.Frontiers in Plant Science,2016(7):347,DOI:10.3389/fpls.2016.00347
[11]李合生.植物生理生化实验原理和技术.北京:高等教育出版社,2000:164-165Li H S.The experiment principle and technique for plant physiology and biochemistry(In Chinese).Beijing:Higher Education Press,2000:164-165
[12]王爱国,罗广华.植物的超氧物自由基与羟胺反应的定量关系.植物生理学通讯,1990(6):55-57Wang A G,Luo G H.Quantitative relation between the reaction of hydroxylamine and superoxide anion radicals in plants(In Chinese).Plant Physiology Communications.1990(6):55-57
[13]林植芳,李双顺,林桂珠,等.衰老叶片和叶绿体中H2O2的积累与膜脂过氧化的关系.植物生理学报,1988,14(1):16-22Lin Z F,Li S S,Lin G Z,et al.The accumulation of hydrogen peroxide in senescencing leaves and chloroplasts in relation to lipid peroxidation(In Chinese).Plant Physiology Journal,1988,14(1):16-22
[14]Dhindsa R S,Plumb-Dhindsa P,Thorpe T A.Leaf senescence:Correlated with increased levels of membrane permeability and lipid peroxidation,and decreased levels of superoxide dismutase and catalase.Journal of Experimental Botany,1981,32(1):93-101
[15]张宪政.作物生理研究方法.北京:农业出版社,1992:197-212Zhang X Z.Study method for crop physiology(In Chinese).Beijing:Agriculture Press,1992:197-212
[16]Nakano Y,Asada K.Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts.Plant and Cell Physiology,1981,22(5):867-880
[17]Foyer C H,Halliwell B.The presence of glutathione and glutathione reductase in chloroplasts:A proposed role in ascorbic acid metabolism.Planta,1976,133(1):21-25
[18]Hossain M A,Nakano Y,Asada K.Monodehy-droascorbate reductase in spinach chloroplasts and its participation in regeneration of ascorbate for scavenging hydrogen peroxide.Plant and Cell Physiology,1984,25(3):385-395
[19]Arakawa N,Tsutsumi K,Sanceda N G,et al.A rapid and sensitive method for the determination of ascorbic acid using4,7-diphenyl-l,10-phenanthroline.Agricultural and Biological Chemistry,1981,45(5):1289-1290
[20]Griffith O W.Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine.Analytical Biochemistry,1980,106:207-212
[21]杜长霞,邵俏赛,樊怀福,等.外源NO对Ca(NO3)2胁迫下黄瓜幼苗生长和抗氧化系统的影响.核农学报,2013,27(6):854-860Du C X,Shao Q S,Fan H F,et al.Effects of exogenous nitric oxide on growth and antioxidant system in cucumber seedlings under Ca(NO3)2 stress(In Chinese).Journal of Nuclear Agricultural Sciences,2013,27(6):854-860
[22]Zhang G W,Liu Z L,Zhou J G,et al.Effects of Ca(NO3)2stress on oxidative damage,antioxidant enzymes activities and polyamine contents in roots of grafted and non-grafted tomato plants.Plant Growth Regulation,2008,56(1):7-19
[23]Wei G P,Yang L F,Zhu Y L,et al.Changes in oxidative damage,antioxidant enzyme activities and polyamine contents in leaves of grafted and non-grafted eggplant seedlings under stress by excess of calcium nitrate.Scientia Horticulturae,2009,120(4):443-451
[24]Nakabayashi R,Saito K.Integrated metabolomics for abiotic stress responses in plants.Current Opinion in Plant Biology,2015,24:10-16
[25]Gallie D R.The role of L-ascorbic acid recycling in responding to environmental stress and in promoting plant growth.Journal of Experimental Botany,2013,64(2):433-443
[26]Pukacka S,Ratajczak E.Antioxidative response of ascorbate-glutathione pathway enzymes and metabolites to desiccation of recalcitrant Acer saccharinum seeds.Journal of Plant Physiology,2006,163(12):1259-1266
[27]Shu S,Gao P,Li L,et al.Abscisic acid-Induced H2O2 accumulation enhances antioxidant capacity in pumpkin-grafted cucumber leaves under Ca(NO3)2 stress.Frontiers in Plant Science,2016(7):1489,DOI:10.3389/fpls.2016.01489
[28]Li L,Shu S,Xu Q,et al.NO accumulation alleviates H2O2-dependent oxidative damage induced by Ca(NO3)2stress in the leaves of pumpkin-grafted cucumber seedlings.Physiologia Plantarum,2017,160(1):33-45
[29]焦娟,王秀峰,杨凤娟,等.外源一氧化氮对硝酸盐胁迫下黄瓜幼苗生长及抗氧化酶活性的影响.应用生态学报,2009,20(12):3009-3014Jiao J,Wang X F,Yang F J,et al.Effects of exogenous NO on the growth and antioxidant enzyme activities of cucumber seedlings under NO3-stress(In Chinese).Chinese Journal of Applied Ecology,2009,20(12):3009-3014
[30]杨全勇,王秀峰,韩宇睿,等.硝普钠对黄瓜幼苗缺镁和硝酸盐胁迫的缓解效应.植物营养与肥料学报,2015,21(5):1269-1278Yang Q Y,Wang X F,Han Y R,et al.Effects of sodium nitroprusside on alleviating magnesium deficiency of cucumber seedlings under NO3-stress(In Chinese).Journal of Plant Nutrition and Fertilizer,2015,21(5):1269-1278
[31]Gro?F,Durner J,Gaupels F.Nitric oxide,antioxidants and prooxidants in plant defence responses.Frontiers in Plant Science,2013(4):419,DOI:10.3389/fpls.2013.00419
[32]刘建新,王鑫,李博萍.外源一氧化氮供体SNP对NaCl胁迫下黑麦草幼苗叶片抗坏血酸-谷胱甘肽循环的影响.草业学报,2010,19(2):82-88Liu J X,Wang X,Li B P.Effects of exogenous nitric oxide donor SNP on AsA-GSH cycle in ryegrass seedlings leaves under salt stress(In Chinese).Acta Prataculturae Sinica,2010,19(2):82-88
[33]Kocsy G,Galiba G,Brunold C.Role of glutathione in adaptation and signalling during chilling and cold acclimation in plants.Physiologia Plantarum,2001,113(2):158-164
[34]樊怀福,郭世荣,段九菊,等.外源NO对NaCl胁迫下黄瓜(Cucumis sativus L.)幼苗生长和谷胱甘肽抗氧化酶系统的影响.生态学报,2008,28(6):2511-2517Fan H F,Guo S R,Duan J J,et al.Effects of nitric oxide on the growth and glutathione dependent antioxidative system in cucumber(Cucumis sativus L.)seedlings under NaCl stress(In Chinese).Acta Ecologica Sinica,2008,28(6):2511-2517
[35]Miyake C,Schreiber U,Hormann H,et al.The FAD-enzyme monodehydroascorbate radical reductase mediates photoproduction of superoxide radicals in spinach thylakoid membranes.Plant and Cell Physiology,1998,39(8):821-829
[2]孙德智,韩晓日,彭靖,等.外源NO对Ca(NO3)2胁迫下番茄幼苗PSII功能及光能分配利用的影响.核农学报,2016,30(12):2451-2459Sun D Z,Han X R,Peng J,et al.Effect of exogenous nitric oxide on PSII function and distribution and utilization of luminous energy in tomato seedlings under stress of Ca(NO3)2(In Chinese).Journal of Nuclear Agricultural Sciences,2016,30(12):2451-2459
[3]Delian E,Bǎdulescu L,Dobrescu A,et al.A brief overview of seed priming benefits in tomato.Romanian Biotechnological Letters,2017,22(3):12505-12513
[4]Santolini J,AndréF,Jeandroz S,et al.Nitric oxide synthase in plants:Where do we stand?Nitric Oxide,2017,63:30-38
[5]Hasanuzzaman M,Nahar K,Alam M M,et al.Exogenous nitric oxide alleviates high temperature induced oxidative stress in wheat(Triticum aestivum L.)seedlings by modulating the antioxidant defense and glyoxalase system.Australian Journal of Crop Science,2012,6(8):1314-1323
[6]Liang L,Deng Y,Sun X,et al.Exogenous nitric oxide pretreatment enhances chilling tolerance of anthurium.Journal of the American Society for Horticultural Science,2018,143(1):3-13
[7]Shan C,Zhou Y,Liu M.Nitric oxide participates in the regulation of the ascorbate-glutathione cycle by exogenous jasmonic acid in the leaves of wheat seedlings under drought stress.Protoplasma,2015,252(5):1397-1405
[8]Esringu A,Aksakal O,Tabay D,et al.Effects of sodium nitroprusside(SNP)pretreatment on UV-B stress tolerance in lettuce(Lactuca sativa L.)seedlings.Environmental Science and Pollution Research,2016,23(1):589-597
[9]Ahmad P,Ahanger M A,Alyemeni M N,et al.Exogenous application of nitric oxide modulates osmolyte metabolism,antioxidants,enzymes of ascorbate-glutathione cycle and promotes growth under cadmium stress in tomato.Protoplasma,2018,255(1):79-93
[10]Ahmad P,Abdel Latef A A,Hashem A,et al.Nitric oxide mitigates salt stress by regulating levels of osmolytes and antioxidant enzymes in chickpea.Frontiers in Plant Science,2016(7):347,DOI:10.3389/fpls.2016.00347
[11]李合生.植物生理生化实验原理和技术.北京:高等教育出版社,2000:164-165Li H S.The experiment principle and technique for plant physiology and biochemistry(In Chinese).Beijing:Higher Education Press,2000:164-165
[12]王爱国,罗广华.植物的超氧物自由基与羟胺反应的定量关系.植物生理学通讯,1990(6):55-57Wang A G,Luo G H.Quantitative relation between the reaction of hydroxylamine and superoxide anion radicals in plants(In Chinese).Plant Physiology Communications.1990(6):55-57
[13]林植芳,李双顺,林桂珠,等.衰老叶片和叶绿体中H2O2的积累与膜脂过氧化的关系.植物生理学报,1988,14(1):16-22Lin Z F,Li S S,Lin G Z,et al.The accumulation of hydrogen peroxide in senescencing leaves and chloroplasts in relation to lipid peroxidation(In Chinese).Plant Physiology Journal,1988,14(1):16-22
[14]Dhindsa R S,Plumb-Dhindsa P,Thorpe T A.Leaf senescence:Correlated with increased levels of membrane permeability and lipid peroxidation,and decreased levels of superoxide dismutase and catalase.Journal of Experimental Botany,1981,32(1):93-101
[15]张宪政.作物生理研究方法.北京:农业出版社,1992:197-212Zhang X Z.Study method for crop physiology(In Chinese).Beijing:Agriculture Press,1992:197-212
[16]Nakano Y,Asada K.Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts.Plant and Cell Physiology,1981,22(5):867-880
[17]Foyer C H,Halliwell B.The presence of glutathione and glutathione reductase in chloroplasts:A proposed role in ascorbic acid metabolism.Planta,1976,133(1):21-25
[18]Hossain M A,Nakano Y,Asada K.Monodehy-droascorbate reductase in spinach chloroplasts and its participation in regeneration of ascorbate for scavenging hydrogen peroxide.Plant and Cell Physiology,1984,25(3):385-395
[19]Arakawa N,Tsutsumi K,Sanceda N G,et al.A rapid and sensitive method for the determination of ascorbic acid using4,7-diphenyl-l,10-phenanthroline.Agricultural and Biological Chemistry,1981,45(5):1289-1290
[20]Griffith O W.Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine.Analytical Biochemistry,1980,106:207-212
[21]杜长霞,邵俏赛,樊怀福,等.外源NO对Ca(NO3)2胁迫下黄瓜幼苗生长和抗氧化系统的影响.核农学报,2013,27(6):854-860Du C X,Shao Q S,Fan H F,et al.Effects of exogenous nitric oxide on growth and antioxidant system in cucumber seedlings under Ca(NO3)2 stress(In Chinese).Journal of Nuclear Agricultural Sciences,2013,27(6):854-860
[22]Zhang G W,Liu Z L,Zhou J G,et al.Effects of Ca(NO3)2stress on oxidative damage,antioxidant enzymes activities and polyamine contents in roots of grafted and non-grafted tomato plants.Plant Growth Regulation,2008,56(1):7-19
[23]Wei G P,Yang L F,Zhu Y L,et al.Changes in oxidative damage,antioxidant enzyme activities and polyamine contents in leaves of grafted and non-grafted eggplant seedlings under stress by excess of calcium nitrate.Scientia Horticulturae,2009,120(4):443-451
[24]Nakabayashi R,Saito K.Integrated metabolomics for abiotic stress responses in plants.Current Opinion in Plant Biology,2015,24:10-16
[25]Gallie D R.The role of L-ascorbic acid recycling in responding to environmental stress and in promoting plant growth.Journal of Experimental Botany,2013,64(2):433-443
[26]Pukacka S,Ratajczak E.Antioxidative response of ascorbate-glutathione pathway enzymes and metabolites to desiccation of recalcitrant Acer saccharinum seeds.Journal of Plant Physiology,2006,163(12):1259-1266
[27]Shu S,Gao P,Li L,et al.Abscisic acid-Induced H2O2 accumulation enhances antioxidant capacity in pumpkin-grafted cucumber leaves under Ca(NO3)2 stress.Frontiers in Plant Science,2016(7):1489,DOI:10.3389/fpls.2016.01489
[28]Li L,Shu S,Xu Q,et al.NO accumulation alleviates H2O2-dependent oxidative damage induced by Ca(NO3)2stress in the leaves of pumpkin-grafted cucumber seedlings.Physiologia Plantarum,2017,160(1):33-45
[29]焦娟,王秀峰,杨凤娟,等.外源一氧化氮对硝酸盐胁迫下黄瓜幼苗生长及抗氧化酶活性的影响.应用生态学报,2009,20(12):3009-3014Jiao J,Wang X F,Yang F J,et al.Effects of exogenous NO on the growth and antioxidant enzyme activities of cucumber seedlings under NO3-stress(In Chinese).Chinese Journal of Applied Ecology,2009,20(12):3009-3014
[30]杨全勇,王秀峰,韩宇睿,等.硝普钠对黄瓜幼苗缺镁和硝酸盐胁迫的缓解效应.植物营养与肥料学报,2015,21(5):1269-1278Yang Q Y,Wang X F,Han Y R,et al.Effects of sodium nitroprusside on alleviating magnesium deficiency of cucumber seedlings under NO3-stress(In Chinese).Journal of Plant Nutrition and Fertilizer,2015,21(5):1269-1278
[31]Gro?F,Durner J,Gaupels F.Nitric oxide,antioxidants and prooxidants in plant defence responses.Frontiers in Plant Science,2013(4):419,DOI:10.3389/fpls.2013.00419
[32]刘建新,王鑫,李博萍.外源一氧化氮供体SNP对NaCl胁迫下黑麦草幼苗叶片抗坏血酸-谷胱甘肽循环的影响.草业学报,2010,19(2):82-88Liu J X,Wang X,Li B P.Effects of exogenous nitric oxide donor SNP on AsA-GSH cycle in ryegrass seedlings leaves under salt stress(In Chinese).Acta Prataculturae Sinica,2010,19(2):82-88
[33]Kocsy G,Galiba G,Brunold C.Role of glutathione in adaptation and signalling during chilling and cold acclimation in plants.Physiologia Plantarum,2001,113(2):158-164
[34]樊怀福,郭世荣,段九菊,等.外源NO对NaCl胁迫下黄瓜(Cucumis sativus L.)幼苗生长和谷胱甘肽抗氧化酶系统的影响.生态学报,2008,28(6):2511-2517Fan H F,Guo S R,Duan J J,et al.Effects of nitric oxide on the growth and glutathione dependent antioxidative system in cucumber(Cucumis sativus L.)seedlings under NaCl stress(In Chinese).Acta Ecologica Sinica,2008,28(6):2511-2517
[35]Miyake C,Schreiber U,Hormann H,et al.The FAD-enzyme monodehydroascorbate radical reductase mediates photoproduction of superoxide radicals in spinach thylakoid membranes.Plant and Cell Physiology,1998,39(8):821-829