葡萄中褪黑素对NaHCO_3胁迫的响应及外源褪黑素缓解NaHCO_3胁迫的作用机制
|
摘要
以一年生‘福客’(‘Frontenac’)葡萄(Vitis sp.)盆栽扦插苗为试材,研究85 mmol·L~(-1) NaHCO_3胁迫处理对葡萄不同器官褪黑素、5-羟色胺和2-羟基褪黑素含量的影响,并进行外源浇灌褪黑素处理,研究外源褪黑素对碱胁迫下有机渗透调节物质和离子含量及离子转运相关基因表达的影响。结果表明,NaHCO_3胁迫后,葡萄根、茎和叶中的褪黑素含量降低,以根和茎中的降低幅度较大,葡萄根系中5-羟色胺的含量显著增加,褪黑素的代谢产物2-羟基褪黑素在根中大幅降低,却在叶片中大量积累;NaHCO_3胁迫下,外源褪黑素显著降低了叶片O_2~(-·)产生速率和H_2O_2含量,增加了有机渗透调节物质可溶性糖、可溶性蛋白和脯氨酸含量,降低了叶片中Na~+含量,但增加了叶片中K~+含量。褪黑素处理提高了NaHCO_3胁迫下葡萄叶片中VvNHXP和VvHKT家族基因的表达。综上,碱胁迫下根系合成褪黑素代谢前体物质5-羟色胺,并向地上部运输,主要在叶片中清除自由基,调节有机渗透调节物质含量,并通过上调离子转运蛋白VvNHXP和VvHKT家族基因的表达,维持胞内离子平衡,进而缓解NaHCO_3对膜系统的伤害。
The one-year-old potted ‘Frontenac' grapevines(Vitis riparia) were used as materials to investigate the effect of 85 mmol·L~(-1) NaHCO_3 stress on melatonin,serotonin and 2-hydroxy melatonin in different organs of grapevines,and exogenous melatonin was irrigated to study the effect of melatonin on the contents of organic osmotic adjustment substances and ions as well as the expression of ion transport-related genes under NaHCO_3 stress.The results show that melatonin content was reduced in leaves and especially in roots and stems under NaHCO_3 stress.In contrast,the content of serotonin in roots were significantly increased under NaHCO_3 treatment,2-hydroxy melatonin in the roots were significantly reduced,and 2-hydroxy melatonin in leaves were increased.Under NaHCO_3 stress,O_2~(-·) production rate and H_2O_2 content were significantly reduced,while contents of organic osmotic adjustment substances,soluble sugar,soluble protein,proline and K+,were increased by exogenous melatonin application;Na~+ content was significantly decreased;the expression levels of VvNHXPand VvHKT family genes were significantly increased in leaves by exogenous melatonin application.In conclusion,serotonin was produced in root and thereafter transported to aerial organs to scavenge free radicals and increase organic osmotic adjustment substances,additionally,the expression of ion transport-related genes,VvNHXP and VvHKT,were up-regulated to maintain ion balance and reduce the membrane injury under NaHCO_3 treatment.
The one-year-old potted ‘Frontenac' grapevines(Vitis riparia) were used as materials to investigate the effect of 85 mmol·L~(-1) NaHCO_3 stress on melatonin,serotonin and 2-hydroxy melatonin in different organs of grapevines,and exogenous melatonin was irrigated to study the effect of melatonin on the contents of organic osmotic adjustment substances and ions as well as the expression of ion transport-related genes under NaHCO_3 stress.The results show that melatonin content was reduced in leaves and especially in roots and stems under NaHCO_3 stress.In contrast,the content of serotonin in roots were significantly increased under NaHCO_3 treatment,2-hydroxy melatonin in the roots were significantly reduced,and 2-hydroxy melatonin in leaves were increased.Under NaHCO_3 stress,O_2~(-·) production rate and H_2O_2 content were significantly reduced,while contents of organic osmotic adjustment substances,soluble sugar,soluble protein,proline and K+,were increased by exogenous melatonin application;Na~+ content was significantly decreased;the expression levels of VvNHXPand VvHKT family genes were significantly increased in leaves by exogenous melatonin application.In conclusion,serotonin was produced in root and thereafter transported to aerial organs to scavenge free radicals and increase organic osmotic adjustment substances,additionally,the expression of ion transport-related genes,VvNHXP and VvHKT,were up-regulated to maintain ion balance and reduce the membrane injury under NaHCO_3 treatment.
引文
Arnao MB,Hernández-Ruiz J(2009a).Protective effect of melatonin against chlorophyll degradation during the senescence of barley leaves.J Pineal Res,46(1):58-63
Arnao MB,Hernández-Ruiz J(2009b).Chemical stress by different agents affects the melatonin content of barley roots.J Pineal Res,46(3):295-299
Arnao MB,Hernández-Ruiz J(2015).Functions of melatonin in plants:a review.J Pineal Res,59(2):133-150
Baxter A,Mittler R,Suzuki N(2014).ROS as key players in plant stress signalling.J Exp Bot,65(5):1229-1240
Byeon Y,Lee HY,Lee K,Park S,Back K(2014).Cellular localization and kinetics of the rice melatonin biosynthetic enzymes SNATand ASMT.J Pineal Res,56(1):107-114
Du YP,Jin XJ,Guo SH,Fu QQ,Zhai H(2015).Effects of different salt and alkali stresses on ion distribution in Red globe/Beta grapevines.Chin J Appl Ecol,26(6):1801-1806(in Chinese with English abstract)[杜远鹏,晋学娟,郭淑华,付晴晴,翟衡(2015).不同盐碱类型胁迫对红地球/贝达葡萄植株离子分布的影响.应用生态学报,26(6):1801-1806]
Dubbels R,Reiter RJ,Klenke E,Goebel A,Schnakenberg E,Ehlers C,Schiwara HW,Schloot W(1995).Melatonin in edible plants identified by radioimmunoassay and by high performance liquid chromatography-mass spectrometry.J Pineal Res,18(1):28-31
Foyer CH,Noctor G(2005).Oxidant and antioxidant signalling in plants:a re-evaluation of the concept of oxidative stress in a physiological context.Plant Cell Environ,28(8):1056-1071
Fu QQ,Geng QW,Zhai H,Du YP(2017).Effects of exogenous melatonin on physiological characteristics of‘Frontenac’grape roots and leaves under Na HCO3 stress.Plant Physiol J,53(2):248-256(in Chinese with English abstract)[付晴晴,耿庆伟,翟衡,杜远鹏(2017).外源褪黑素对Na HCO3胁迫下‘福客’葡萄根系和叶片生理特性的影响.植物生理学报,53(2):248-256]
Galano A,Tan DX,Reiter RJ(2011).Melatonin as a natural ally against oxidative stress:a physicochemical examination.J Pineal Res,51(1):1-16
Gao QH,Guo YY,Wu Y,Jia SS(2017).Alleviation effects of MT and Ca2+on melon seedlings under salt stress.Chin J Appl Ecol,28(6):1925-1931(in Chinese with English abstract)[高青海,郭远远,吴燕,贾双双(2017).盐胁迫下外源褪黑素和Ca2+对甜瓜幼苗的缓解效应.应用生态学报,28(6):1925-1931]
Gao QH,Jia SS,Miao YM,Lu XM,Li HM(2016).Effects of exogenous melatonin on nitrogen metabolism and osmotic adjustment substances of melon seedlings under sub-low temperature.Chin J Appl Ecol,27(2):519-524(in Chinese with English abstract)[高青海,贾双双,苗永美,陆晓民,李慧敏(2016).亚低温条件下外源褪黑素对甜瓜幼苗氮代谢及渗透调节物质的影响.应用生态学报27(2):519-524]
Guo R,Li F,Zhou J,Li HR,Xia X,Liu Q(2016).Eco-physiological responses of linseed(Linum usitatissimum)to salt and alkali stresses.Chin J Plant Ecol,40(1):69-79(in Chinese with English abstract)[郭瑞,李峰,周际,李昊儒,夏旭,刘琪(2016).亚麻响应盐、碱胁迫的生理特征.植物生态学报,40(1):69-79]
Koyama FC,Carvalho TLG,Alves E,da Silva HB,de Azevedo MF,Hemerly AS,Garcia CRS(2013).The structurally related auxin and melatonin tryptophan-derivatives and their roles in Arabidopsis thaliana and in the human malaria parasite Plasmodium falciparum.J Eukaryot Microbiol,60(6):646-651
Li C,Wang P,Wei Z,Liang D,Liu C,Yin L,Jia D,Fu M,Ma F(2012).The mitigation effects of exogenous melatonin on salinity-induced stress in Malus hupehensis.J Pineal Res,53(3):298-306
Li Y,Zhang Y,Feng F,Liang D,Cheng L,Ma F,Shi S(2010).Overexpression of a Malus vacuolar Na+/H+antiporter gene(Md N-HX1)in apple rootstock M.26 and its influence on salt tolerance.Plant Cell Tiss Organ Cult,102(3):337-345
Li YN,Yuan SX,Xu LF,Yang PP,Feng YY,Hir YP,Ming J(2016).The physiological response of lily to Na HCO3 stress.Acta Hortic Sin,43(6):1126-1140(in Chinese with English abstract)[李雅男,袁素霞,徐雷锋,杨盼盼,冯亚言,Hir YP,明军(2016).百合在Na HCO3胁迫下的生理响应机制.园艺学报,43(6):1126-1140]
Li ZG,Gong M(2005).Improvement of measurement method for superoxide anion radical in plant.Acta Bot Yunnan,27(2):211-216(in Chinese with English abstract)[李忠光,龚明(2005).植物中超氧阴离子自由基测定方法的改进.云南植物研究,27(2):211-216]
Lin Z,Li S,Lin G,Guo J(1988).The accumulation of hydrogen peroxide in senescing leaves and chloroplasts in relation to lipid peroxidation.Acta Phytophysiol Sin,14(1):16-22(in Chinese)[林植芳,李双顺,林桂珠,郭俊彦(1988).衰老叶片和叶绿体中H2O2的累积与膜脂过氧化的关系.植物生理学报,14(1):16-22]
Liu N,Jin Z,Wang S,Gong B,Wen D,Wang X,Wei M,Shi Q(2015).Sodic alkaline stress mitigation with exogenous melatonin involves reactive oxygen metabolism and ion homeostasis in tomato.Sci Hortic,181(2):18-25
Livak K,Schmittgen TD(2001).Analysis of relative gene expression data using real-time quantitative PCR and the 2-??CT method.Methods,25(4):402-408
Meloni DA,Oliva MA,Martinez CA,Cambraia J(2003).Photosynthesis and activity of superoxide dismutase,peroxidase and glutathione reductase in cotton under salt stress.Environ Exp Bot,49(1):69-76
Munns R,Tester M(2008).Mechanisms of salinity tolerance.Annu Rev Plant Biol,59:651-681
Nazar R,Iqbal N,Syeed S,Khan NA(2011).Salicylic acid alleviates decreases in photosynthesis under salt stress by enhancing nitrogen and sulfur assimilation and antioxidant metabolism differentially in two mungbean cultivars.J Plant Physiol,168(8):807-815
Parida AK,Das AB(2005).Salt tolerance and salinity effects on plants:a review.Ecotoxicol Environ,60(3):324-349
Shida CS,Castrucci AML,Lamy-Freund MT(1994).Lamy-Freund MT.High melatonin solubility in aqueous medium.J Plant Physiol,16(4):198-201
Turk H,Erdal S,Genisel M,Atici O,Demir Y,Yanmis D(2014).The regulatory effect of melatonin on physiological,biochemical and molecular parameters in cold-stressed wheat seedlings.Plant Growth Regul,74(2):139-152
Verslues PE,Agarwal M,Katiyar-Agarwal S,Zhu J,Zhu JK(2006).Methods and concepts in quantifying resistance to drought,salt and freezing,abiotic stresses that affect plant water status.Plant J,45(4):523-539
Xu F,Zhou H,Guo Z,Yu HA,Yuan Y,Gong Z,Wang Y(2013).The melatonin and its resistance to stress in plants.Genom Appl Biol,32(2):260-266(in Chinese with English abstract)[徐芳,周海鹏,郭早霞,于宏安,袁玉川,巩智刚,王玉华(2013).植物褪黑素及其抗逆性研究.基因组学与应用生物学,32(2):260-266]
Yin L(2014).The establishment and application of the method for detection of melatonin in mulberry and mulberry wine(Master’s thesis).Beijing:China Agricultural University(in Chinese with English abstract)[尹丽媛(2014).桑葚果实及桑葚酒中褪黑素检测方法的建立与应用研究(硕士论文).北京:中国农业大学]
Zhang N,Zhang H,Yang R,Huang Y,Guo Y(2012).The advances on functions study of melatonin in plants.Chin Agr Sci Bull,28(9):16-20(in Chinese with English abstract)[张娜,张海军,杨荣超,黄韫宇,郭仰东(2012).褪黑素在植物中的功能研究进展.中国农学通报,28(9):16-20]
Zhao S,Shi G,Dong X(2002).Techniques of Plant Physiological Experiment.Beijing:China Agricultural Science and Technology Press(in Chinese)[赵世杰,史国安,董新纯(2002).植物生理学实验指导.北京:中国农业科学技术出版社]
Zhu JK(2003).Regulation of ion homeostasis under salt stress.Curr Opin Plant Biol,6(5):441-445
Zuo B,Zheng X,He P,Wang L,Lei Q,Feng C,Zhou J,Li Q,Han Z,Kong J(2014).Overexpression of Mz ASMT improves melatonin production and enhances drought tolerance in transgenic Arabidopsis thaliana plants.J Pineal Res,57(4):408-417
Arnao MB,Hernández-Ruiz J(2009b).Chemical stress by different agents affects the melatonin content of barley roots.J Pineal Res,46(3):295-299
Arnao MB,Hernández-Ruiz J(2015).Functions of melatonin in plants:a review.J Pineal Res,59(2):133-150
Baxter A,Mittler R,Suzuki N(2014).ROS as key players in plant stress signalling.J Exp Bot,65(5):1229-1240
Byeon Y,Lee HY,Lee K,Park S,Back K(2014).Cellular localization and kinetics of the rice melatonin biosynthetic enzymes SNATand ASMT.J Pineal Res,56(1):107-114
Du YP,Jin XJ,Guo SH,Fu QQ,Zhai H(2015).Effects of different salt and alkali stresses on ion distribution in Red globe/Beta grapevines.Chin J Appl Ecol,26(6):1801-1806(in Chinese with English abstract)[杜远鹏,晋学娟,郭淑华,付晴晴,翟衡(2015).不同盐碱类型胁迫对红地球/贝达葡萄植株离子分布的影响.应用生态学报,26(6):1801-1806]
Dubbels R,Reiter RJ,Klenke E,Goebel A,Schnakenberg E,Ehlers C,Schiwara HW,Schloot W(1995).Melatonin in edible plants identified by radioimmunoassay and by high performance liquid chromatography-mass spectrometry.J Pineal Res,18(1):28-31
Foyer CH,Noctor G(2005).Oxidant and antioxidant signalling in plants:a re-evaluation of the concept of oxidative stress in a physiological context.Plant Cell Environ,28(8):1056-1071
Fu QQ,Geng QW,Zhai H,Du YP(2017).Effects of exogenous melatonin on physiological characteristics of‘Frontenac’grape roots and leaves under Na HCO3 stress.Plant Physiol J,53(2):248-256(in Chinese with English abstract)[付晴晴,耿庆伟,翟衡,杜远鹏(2017).外源褪黑素对Na HCO3胁迫下‘福客’葡萄根系和叶片生理特性的影响.植物生理学报,53(2):248-256]
Galano A,Tan DX,Reiter RJ(2011).Melatonin as a natural ally against oxidative stress:a physicochemical examination.J Pineal Res,51(1):1-16
Gao QH,Guo YY,Wu Y,Jia SS(2017).Alleviation effects of MT and Ca2+on melon seedlings under salt stress.Chin J Appl Ecol,28(6):1925-1931(in Chinese with English abstract)[高青海,郭远远,吴燕,贾双双(2017).盐胁迫下外源褪黑素和Ca2+对甜瓜幼苗的缓解效应.应用生态学报,28(6):1925-1931]
Gao QH,Jia SS,Miao YM,Lu XM,Li HM(2016).Effects of exogenous melatonin on nitrogen metabolism and osmotic adjustment substances of melon seedlings under sub-low temperature.Chin J Appl Ecol,27(2):519-524(in Chinese with English abstract)[高青海,贾双双,苗永美,陆晓民,李慧敏(2016).亚低温条件下外源褪黑素对甜瓜幼苗氮代谢及渗透调节物质的影响.应用生态学报27(2):519-524]
Guo R,Li F,Zhou J,Li HR,Xia X,Liu Q(2016).Eco-physiological responses of linseed(Linum usitatissimum)to salt and alkali stresses.Chin J Plant Ecol,40(1):69-79(in Chinese with English abstract)[郭瑞,李峰,周际,李昊儒,夏旭,刘琪(2016).亚麻响应盐、碱胁迫的生理特征.植物生态学报,40(1):69-79]
Koyama FC,Carvalho TLG,Alves E,da Silva HB,de Azevedo MF,Hemerly AS,Garcia CRS(2013).The structurally related auxin and melatonin tryptophan-derivatives and their roles in Arabidopsis thaliana and in the human malaria parasite Plasmodium falciparum.J Eukaryot Microbiol,60(6):646-651
Li C,Wang P,Wei Z,Liang D,Liu C,Yin L,Jia D,Fu M,Ma F(2012).The mitigation effects of exogenous melatonin on salinity-induced stress in Malus hupehensis.J Pineal Res,53(3):298-306
Li Y,Zhang Y,Feng F,Liang D,Cheng L,Ma F,Shi S(2010).Overexpression of a Malus vacuolar Na+/H+antiporter gene(Md N-HX1)in apple rootstock M.26 and its influence on salt tolerance.Plant Cell Tiss Organ Cult,102(3):337-345
Li YN,Yuan SX,Xu LF,Yang PP,Feng YY,Hir YP,Ming J(2016).The physiological response of lily to Na HCO3 stress.Acta Hortic Sin,43(6):1126-1140(in Chinese with English abstract)[李雅男,袁素霞,徐雷锋,杨盼盼,冯亚言,Hir YP,明军(2016).百合在Na HCO3胁迫下的生理响应机制.园艺学报,43(6):1126-1140]
Li ZG,Gong M(2005).Improvement of measurement method for superoxide anion radical in plant.Acta Bot Yunnan,27(2):211-216(in Chinese with English abstract)[李忠光,龚明(2005).植物中超氧阴离子自由基测定方法的改进.云南植物研究,27(2):211-216]
Lin Z,Li S,Lin G,Guo J(1988).The accumulation of hydrogen peroxide in senescing leaves and chloroplasts in relation to lipid peroxidation.Acta Phytophysiol Sin,14(1):16-22(in Chinese)[林植芳,李双顺,林桂珠,郭俊彦(1988).衰老叶片和叶绿体中H2O2的累积与膜脂过氧化的关系.植物生理学报,14(1):16-22]
Liu N,Jin Z,Wang S,Gong B,Wen D,Wang X,Wei M,Shi Q(2015).Sodic alkaline stress mitigation with exogenous melatonin involves reactive oxygen metabolism and ion homeostasis in tomato.Sci Hortic,181(2):18-25
Livak K,Schmittgen TD(2001).Analysis of relative gene expression data using real-time quantitative PCR and the 2-??CT method.Methods,25(4):402-408
Meloni DA,Oliva MA,Martinez CA,Cambraia J(2003).Photosynthesis and activity of superoxide dismutase,peroxidase and glutathione reductase in cotton under salt stress.Environ Exp Bot,49(1):69-76
Munns R,Tester M(2008).Mechanisms of salinity tolerance.Annu Rev Plant Biol,59:651-681
Nazar R,Iqbal N,Syeed S,Khan NA(2011).Salicylic acid alleviates decreases in photosynthesis under salt stress by enhancing nitrogen and sulfur assimilation and antioxidant metabolism differentially in two mungbean cultivars.J Plant Physiol,168(8):807-815
Parida AK,Das AB(2005).Salt tolerance and salinity effects on plants:a review.Ecotoxicol Environ,60(3):324-349
Shida CS,Castrucci AML,Lamy-Freund MT(1994).Lamy-Freund MT.High melatonin solubility in aqueous medium.J Plant Physiol,16(4):198-201
Turk H,Erdal S,Genisel M,Atici O,Demir Y,Yanmis D(2014).The regulatory effect of melatonin on physiological,biochemical and molecular parameters in cold-stressed wheat seedlings.Plant Growth Regul,74(2):139-152
Verslues PE,Agarwal M,Katiyar-Agarwal S,Zhu J,Zhu JK(2006).Methods and concepts in quantifying resistance to drought,salt and freezing,abiotic stresses that affect plant water status.Plant J,45(4):523-539
Xu F,Zhou H,Guo Z,Yu HA,Yuan Y,Gong Z,Wang Y(2013).The melatonin and its resistance to stress in plants.Genom Appl Biol,32(2):260-266(in Chinese with English abstract)[徐芳,周海鹏,郭早霞,于宏安,袁玉川,巩智刚,王玉华(2013).植物褪黑素及其抗逆性研究.基因组学与应用生物学,32(2):260-266]
Yin L(2014).The establishment and application of the method for detection of melatonin in mulberry and mulberry wine(Master’s thesis).Beijing:China Agricultural University(in Chinese with English abstract)[尹丽媛(2014).桑葚果实及桑葚酒中褪黑素检测方法的建立与应用研究(硕士论文).北京:中国农业大学]
Zhang N,Zhang H,Yang R,Huang Y,Guo Y(2012).The advances on functions study of melatonin in plants.Chin Agr Sci Bull,28(9):16-20(in Chinese with English abstract)[张娜,张海军,杨荣超,黄韫宇,郭仰东(2012).褪黑素在植物中的功能研究进展.中国农学通报,28(9):16-20]
Zhao S,Shi G,Dong X(2002).Techniques of Plant Physiological Experiment.Beijing:China Agricultural Science and Technology Press(in Chinese)[赵世杰,史国安,董新纯(2002).植物生理学实验指导.北京:中国农业科学技术出版社]
Zhu JK(2003).Regulation of ion homeostasis under salt stress.Curr Opin Plant Biol,6(5):441-445
Zuo B,Zheng X,He P,Wang L,Lei Q,Feng C,Zhou J,Li Q,Han Z,Kong J(2014).Overexpression of Mz ASMT improves melatonin production and enhances drought tolerance in transgenic Arabidopsis thaliana plants.J Pineal Res,57(4):408-417