过氧化氢提高燕麦幼苗耐碱性的活性氧代谢和渗透调节
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摘要
为探明过氧化氢(H_2O_2)提高燕麦耐碱性的作用,以‘定莜6号’幼苗为材料,采用珍珠岩栽培方法,在幼苗三叶一心期根部浇灌75mmol·L~(-1) NaHCO_3或添加二甲基硫脲(DMTU,H_2O_2淬灭剂)或抗坏血酸(ASA,H_2O_2清除剂)模拟碱胁迫,通过叶面喷施0.01mmol·L~(-1) H_2O_2来观测H_2O_2对碱胁迫下幼苗生长、活性氧代谢和渗透溶质积累的影响。结果表明:喷施H_2O_2能够缓解NaHCO_3胁迫对燕麦幼苗生长的抑制,降低幼苗叶片O_2·~-、H_2O_2和丙二醛含量,提高超氧化物歧化酶、抗坏血酸过氧化物酶等抗氧化酶活性和非酶抗氧化剂抗坏血酸、谷胱甘肽含量;使过氧化氢酶、过氧化物酶活性及渗透溶质可溶性糖、游离氨基酸和脯氨酸含量显著降低,有机酸和叶绿素含量明显提高,而可溶性蛋白质含量则变化不大;添加DMTU和ASA后有效逆转了喷施H_2O_2对NaHCO_3胁迫燕麦生长受抑和生理响应的调节。采用主成分和隶属函数分析显示,喷施H_2O_2显著提高了NaHCO_3胁迫下燕麦幼苗的综合评价值,添加DMTU和ASA完全或部分逆转了H_2O_2的作用。因此,外源H_2O_2是通过调控活性氧代谢和渗透调节来缓解碱胁迫导致的氧化伤害和生长抑制,从而增强燕麦幼苗的耐碱性。
Hydrogen peroxide(H_2O_2)is a signaling molecule with diverse physiological functions in plants.The objective of this study was to determine whether exogenous H_2O_2 could improve the alkali tolerance of oat(Avena nuda)seedlings.Seedlings of oat cv.Dingyou No.6 at the three-leaf stage were irrigated with 75 mmol·L~(-1) NaHCO_3 solution containing dimethylthiourea(DMTU,aquencher of H_2O_2)or ascorbate(ASA,a scavenger of H_2O_2),and the leaves were sprayed with 0.01 mmol·L-1 H_2O_2.Seedlings irrigated with water served as the control.After 7 days of treatment,seedling growth,chlorophyll content,active oxygen metabolism,and osmotic accumulation in leaves were measured.The results showed that treatments with exogenous H_2O_2 remarkably relieved the growth inhibition caused by NaHCO_3 stress,and significantly decreased the contents of O2·-,H_2O_2,malondialdehyde,soluble sugars,free amino acids,proline,and activities of catalase and peroxidase.Treatments with exogenous H_2O_2 also increased the activities of superoxide dismutase and ascorbate peroxidase,and increased the contents of ascorbic acid,glutathione,organic acids,and chlorophyll in leaves of oat seedlings.However,none of the treatments affected the soluble protein content.The regulating effects of exogenous H_2O_2 on alkali stress could be reversed by treatment with DMTU or ASA.A comprehensive evaluation based on principal components and subordinate function analyses revealed that foliar spraying with H_2O_2 could improve the alkali stress adaptability of oat seedlings,and the promoting effect of H_2O_2 could be reversed by DMTU or ASA.Together,the results showed that H_2O_2 can increase alkali resistance in oat seedlings by regulating active oxygen metabolism and osmotic accumulation in leaves oat plants under alkali stress.
Hydrogen peroxide(H_2O_2)is a signaling molecule with diverse physiological functions in plants.The objective of this study was to determine whether exogenous H_2O_2 could improve the alkali tolerance of oat(Avena nuda)seedlings.Seedlings of oat cv.Dingyou No.6 at the three-leaf stage were irrigated with 75 mmol·L~(-1) NaHCO_3 solution containing dimethylthiourea(DMTU,aquencher of H_2O_2)or ascorbate(ASA,a scavenger of H_2O_2),and the leaves were sprayed with 0.01 mmol·L-1 H_2O_2.Seedlings irrigated with water served as the control.After 7 days of treatment,seedling growth,chlorophyll content,active oxygen metabolism,and osmotic accumulation in leaves were measured.The results showed that treatments with exogenous H_2O_2 remarkably relieved the growth inhibition caused by NaHCO_3 stress,and significantly decreased the contents of O2·-,H_2O_2,malondialdehyde,soluble sugars,free amino acids,proline,and activities of catalase and peroxidase.Treatments with exogenous H_2O_2 also increased the activities of superoxide dismutase and ascorbate peroxidase,and increased the contents of ascorbic acid,glutathione,organic acids,and chlorophyll in leaves of oat seedlings.However,none of the treatments affected the soluble protein content.The regulating effects of exogenous H_2O_2 on alkali stress could be reversed by treatment with DMTU or ASA.A comprehensive evaluation based on principal components and subordinate function analyses revealed that foliar spraying with H_2O_2 could improve the alkali stress adaptability of oat seedlings,and the promoting effect of H_2O_2 could be reversed by DMTU or ASA.Together,the results showed that H_2O_2 can increase alkali resistance in oat seedlings by regulating active oxygen metabolism and osmotic accumulation in leaves oat plants under alkali stress.
引文
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[15]Chen J X,Wang X F.Plant Physiology Experiment Instruction.Guangzhou:South China University of Technology Press,2006:68-77.陈建勋,王晓峰.植物生理学实验指导.广州:华南理工大学出版社,2006:68-77.
[16]Wang L,Duan X Y,Tang M,et al.Comprehensive evaluation of physiol ogical indices of cold resistance in cymbidium under lanthanum treatment.Plant Physiology Journal,2014,50(8):1177-1183.王玲,段晓宇,唐敏,等.镧处理下大花蕙兰耐寒性生理指标的综合评价.植物生理学报,2014,50(8):1177-1183.
[17]Fedina S,Nedeva D,Cicek N.Pre-treatment with H2O2induces salt tolerance in Barley seedlings.Biologia Plantarum,2009,53(2):321-324.
[18]Gao Y,Guo Y K,Lin S H,et al.Hydrogen peroxide pretreatment alters the activity of antioxidant enzymes and protects chloroplast ultrastructure in heat-stressed cucumber leaves.Scientia Horticulturae,2010,126(1):20-26.
[19]Zhang B,Zhang H G.Regulation of exogenous hydrogen peroxide on wheat seedling salinity tolerance.Acta Botanica BorealiOccidentalia Sinica,2007,27(12):2491-2495.张波,张怀刚.外源H2O2对小麦幼苗耐盐性的调节作用.西北植物学报,2007,27(12):2491-2495.
[20]Lu W,Li S,Li G F,et al.Effect of H2O2on stability of rice Rubisco.Plant Physiology Journal,2002,38(3):213-216.陆巍,李森,李国富,等.H2O2对水稻Rubisco稳定性的影响.植物生理学报,2002,38(3):213-216.
[21]Azevedo N A D,Prisco J T,Enéas-Filho J,et al.Hydrogen peroxide pre-treatment induces salt-stress acclimation in maize plants.Journal of Plant Physiology,2005,162(10):1114-1122.
[2]Yang C,Shi D,Wang D.Comparative effects of salt stress and alkali stress on growth,osmotic adjustment and ionic balance of an alkali resistant halophyte Suaeda glauca Bge.Plant Growth Regulation,2008,56(2):179-190.
[3]Gong B,Wen D,VandenLangenberg K,et al.Comparative effects of NaCl and NaHCO3stress on photosynthetic parameters,nutrient metabolism,and the antioxidant system in tomato leaves.Scientia Horticulturae,2013,157(3):1-12.
[4]Drzikova B,Dongowski G,Gebhardt E.Dietary fibre-rich oat-based products affect serum lipids,microbiota,formation of short-chain fatty acids and steroids in rats.British Journal of Nutrition,2005,94(6):1012-1025.
[5]Liu J X,Wang J C,Wang R J,et al.Exogenous nitric oxide elevated alkali tolerance of Avena nudaseedlings.Acta Prataculturae Sinica,2015,24(8):110-117.刘建新,王金成,王瑞娟,等.外源一氧化氮提高裸燕麦幼苗的耐碱性.草业学报,2015,24(8):110-117.
[6]Uchida A,Jagendorf A T,Hibino T,et al.Effects of hydrogen peroxide and nitric oxide on both salt and heat stress tolerance in rice.Plant Science,2002,163(3):515-523.
[7]An G Y,Li B Z,Wu G L,et al.H2O2Could act as root source signal to mediate stomatal closure induced by salt stress of Vicia faba L.Plant Physiology Journal,2012,48(3):265-271.安国勇,李保珠,武桂丽,等.H2O2作为根源信号介导盐胁迫诱导的蚕豆气孔关闭反应.植物生理学报,2012,48(3):265-271.
[8]Abdul W,Mubaraka P,Sadia G,et al.Pretreatment of seed with H2O2improves salt tolerance of wheat seedlings by alleviation of oxidative damage and expression of stress proteins.Journal of Plant Physiology,2007,164(3):283-294.
[9]Liu Z J,Guo Y K,Lin S H,et al.Effects of exogenous hydrogen peroxide on ultra structure of chloroplasts and activities of antioxidant enzymes in greenhouse-ecotype cucumber under drought stress.Acta Horticulturae Sinica,2009,36(8):1140-1146.刘忠静,郭延奎,林少航,等.外源过氧化氢对干旱胁迫下温室黄瓜叶绿体超微结构和抗氧化酶的影响.园艺学报,2009,36(8):1140-1146.
[10]Li X D,Hou L X,Liu X,et al.Involvement of H2O2in regulating the expression of VvIPK2in response to low temperature stress in leaves of Vitis.Acta Horticulturae Sinica,2011,38(6):1052-1062.李希东,侯丽霞,刘新,等.H2O2与葡萄VvIPK2基因表达及其低温胁迫响应的关系.园艺学报,2011,38(6):1052-1062.
[11]Bai X J,Liu L J,Zhang C H,et al.Effect of H2O2pretreatment on Cd tolerance of different rice cultivars.Chinese Journal of Rice Science,2010,24(4):391-397.白晓娟,刘丽娟,张春华,等.H2O2预处理对不同水稻品种Cd耐性的影响.中国水稻科学,2010,24(4):391-397.
[12]Gao J F.Plant Physiology Experiment Guide.Beijing:Higher Education Press,2006:74-77,199-222.高俊凤.植物生理学实验指导.北京:高等教育出版社,2006:74-77,199-222.
[13]Sergiev I,Alexieva V,Karanov E.Effect of spermine,atrazine and combination between them on some endogenous protective systems and stress markers in plants.Comptes Rendus de I’Academie Bulgare des Sciences,1997,51(2):121-124.
[14]Li H S.The Experiment Principle and Technique on Plant Physiology and Biochemistry.Beijing:Higher Education Press,2000:184-196,258-261.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,2000:184-196,258-261.
[15]Chen J X,Wang X F.Plant Physiology Experiment Instruction.Guangzhou:South China University of Technology Press,2006:68-77.陈建勋,王晓峰.植物生理学实验指导.广州:华南理工大学出版社,2006:68-77.
[16]Wang L,Duan X Y,Tang M,et al.Comprehensive evaluation of physiol ogical indices of cold resistance in cymbidium under lanthanum treatment.Plant Physiology Journal,2014,50(8):1177-1183.王玲,段晓宇,唐敏,等.镧处理下大花蕙兰耐寒性生理指标的综合评价.植物生理学报,2014,50(8):1177-1183.
[17]Fedina S,Nedeva D,Cicek N.Pre-treatment with H2O2induces salt tolerance in Barley seedlings.Biologia Plantarum,2009,53(2):321-324.
[18]Gao Y,Guo Y K,Lin S H,et al.Hydrogen peroxide pretreatment alters the activity of antioxidant enzymes and protects chloroplast ultrastructure in heat-stressed cucumber leaves.Scientia Horticulturae,2010,126(1):20-26.
[19]Zhang B,Zhang H G.Regulation of exogenous hydrogen peroxide on wheat seedling salinity tolerance.Acta Botanica BorealiOccidentalia Sinica,2007,27(12):2491-2495.张波,张怀刚.外源H2O2对小麦幼苗耐盐性的调节作用.西北植物学报,2007,27(12):2491-2495.
[20]Lu W,Li S,Li G F,et al.Effect of H2O2on stability of rice Rubisco.Plant Physiology Journal,2002,38(3):213-216.陆巍,李森,李国富,等.H2O2对水稻Rubisco稳定性的影响.植物生理学报,2002,38(3):213-216.
[21]Azevedo N A D,Prisco J T,Enéas-Filho J,et al.Hydrogen peroxide pre-treatment induces salt-stress acclimation in maize plants.Journal of Plant Physiology,2005,162(10):1114-1122.