外源cGMP调控盐胁迫下黑麦草种子萌发机制
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摘要
c GMP(cyclic guanosine-3',5'-monophosphate)是一类环化核苷酸,能响应植物非生物胁迫的信号分子。以c GMP的膜透性类似物8-Br-cGMP为供体,研究c GMP对盐胁迫下黑麦草种子萌发及生理指标的影响。通过计算萌发指标、测定种子萌发过程中各项生理指标,探讨c GMP对黑麦草种子萌发时耐盐机制的作用。结果表明:低浓度(50 mmol/L)盐胁迫能促进黑麦草种子的萌发,随着盐浓度(100、150、200、250 mmol/L)的增加黑麦草种子的萌发率逐渐降低甚至抑制其萌发。而加入20μmol/L c GMP能缓解100 mmol/L Na Cl胁迫对黑麦草种子造成的伤害,使黑麦草种子的发芽率、发芽势、发芽指数、活力指数分别提高了7.4%、133.3%、52.1%、104.2%。此外,研究发现加入20μmol/Lc GMP能够促进100 mmol/L Na Cl盐胁迫下黑麦草萌发期根的生长,比单独100 mmol/L Na Cl盐胁迫处理下的黑麦草植株全长、根长、叶长分别提高了2.5、2.8倍和2.6倍,种子在萌发过程中可溶性糖、可溶性蛋白含量、淀粉酶活性,脯氨酸含量分别提高了47.9%、15.7%、94.3%、117.4%,而淀粉含量、MDA含量、电导率,O_2~-产生速率分别降低了40.9%、128.7%、88.6%、211.9%。说明20μmol/Lc GMP通过提高可溶性糖,可溶性蛋白含量及脯氨酸含量,同时减少MDA、O_2~-的产生,以此缓解盐胁迫对黑麦草种子的伤害、同时促进淀粉水解,从而加速种子萌发。
c GMP(cyclic guanosine-3',5'-monophosphate)is a type of cyclic nucleotide and signaling molecule that can be responsive to plant abiotic stress.Using c GMP membrane permeability analog 8-Br-cGMP as a donor,this study investigated the effects of c GMP on seed germination and physiological indicators of ryegrass under salt stress.The effects of c GMP on the salt tolerance mechanism during the germination of ryegrass were determined through calculating the germination index and determining the physiological indicators in the process of seed germination.The results showed that a low concentration(50 mmol/L)of salt stress could promote the germination of ryegrass seeds.With increasing salt concentrations(100,150,200,and 250 mmol/L),the germination rate of ryegrass seeds gradually decreased and was even inhibited.Adding 20μmol/L c GMP can relieve the damage caused by 100 mmol/L Na Cl stress on ryegrass seed,increasing the germination rate,germination potential,germination index,and vigor index of ryegrass seed by 7.4%,133.3%,52.1%,and 104.2%,respectively.In addition,compared to the 100 mmol/L Na Cl stress condition alone,the growth of roots of ryegrass could be promoted and the total length,root length,and leaf length of ryegrass plants increased,by 2.5,2.8,and 2.6 times,respectively,when 20μmol/L c GMP was added.The soluble sugar,soluble protein content,amylase activity,and proline content of seeds increased by 47.9%,15.7%,94.3%,and 117.4%,respectively,while starch content,MDA content,conductivity,and O_2~-.production rate were reduced by 40.9%,128.7%,88.6%,and 211.9%,respectively.The results showed that 20μmol/L c GMP could increase the contents of soluble sugar,soluble protein,and proline,and reduce the production of MDA and O_2~-,so as to relieve the damage caused by salt stress on ryegrass seed and promote the hydrolysis of starch,which accelerated seed germination.
c GMP(cyclic guanosine-3',5'-monophosphate)is a type of cyclic nucleotide and signaling molecule that can be responsive to plant abiotic stress.Using c GMP membrane permeability analog 8-Br-cGMP as a donor,this study investigated the effects of c GMP on seed germination and physiological indicators of ryegrass under salt stress.The effects of c GMP on the salt tolerance mechanism during the germination of ryegrass were determined through calculating the germination index and determining the physiological indicators in the process of seed germination.The results showed that a low concentration(50 mmol/L)of salt stress could promote the germination of ryegrass seeds.With increasing salt concentrations(100,150,200,and 250 mmol/L),the germination rate of ryegrass seeds gradually decreased and was even inhibited.Adding 20μmol/L c GMP can relieve the damage caused by 100 mmol/L Na Cl stress on ryegrass seed,increasing the germination rate,germination potential,germination index,and vigor index of ryegrass seed by 7.4%,133.3%,52.1%,and 104.2%,respectively.In addition,compared to the 100 mmol/L Na Cl stress condition alone,the growth of roots of ryegrass could be promoted and the total length,root length,and leaf length of ryegrass plants increased,by 2.5,2.8,and 2.6 times,respectively,when 20μmol/L c GMP was added.The soluble sugar,soluble protein content,amylase activity,and proline content of seeds increased by 47.9%,15.7%,94.3%,and 117.4%,respectively,while starch content,MDA content,conductivity,and O_2~-.production rate were reduced by 40.9%,128.7%,88.6%,and 211.9%,respectively.The results showed that 20μmol/L c GMP could increase the contents of soluble sugar,soluble protein,and proline,and reduce the production of MDA and O_2~-,so as to relieve the damage caused by salt stress on ryegrass seed and promote the hydrolysis of starch,which accelerated seed germination.
引文
[1] Marondedze C,Wong A,Thomas L,Irving H,Gehring C. Cyclic nucleotide monophosphates in plants and plant signaling//Seifert R,ed. Noncanonical Cyclic Nucleotides. Cham:Springer,2015,238:87-103.
[2] Irving H R,Gehring C. Molecular methods for the study of signal transduction in plants//Gehring C,ed. Cyclic Nucleotide Signaling in Plants.Totowa,NJ:Humana Press,2013,1016:1-11.
[3] Pagnussat G C,Lanteri M L,Lamattina L. Nitric oxide and cyclic GMP are messengers in the indole acetic acid-induced adventitious rooting process. Plant Physiology,2013,132(3):1241-1248.
[4] Meier S,Gehring C. Emerging roles in plant biotechnology for the second messenger c GMP-guanosine 3',5'-cyclic monophosphate. African Journal of Biotechnology,2006,5(19):1687-1692.
[5] Isner J C,Maathuis F J M. Measurement of cellular c GMP in plant cells and tissues using the endogenous fluorescent reporter Flinc G. Plant Journal,2011,65(2):329-334.
[6] Turek I,Gehring C. The plant natriuretic peptide receptor is a guanylyl cyclase and enables c GMP-dependent signaling. Plant Molecular Biology,2016,91(3):275-286.
[7] Durner J,Wendehenne D,Klessig D F. Defense gene induction in tobacco by nitric oxide,cyclic GMP,and cyclic ADP-ribose. Proceedings of the National Academy of Sciences of the United States of America,1998,95(17):10328-10333.
[8] Rubio F,Flores P,Navarro J M,Martínez V. Effects of Ca2+,K+and c GMP on Na+uptake in pepper plants. Plant Science,2003,165(5):1043-1049.
[9]刘春英,张学映,朱体超,陈光蓉,郑章云.不同黑麦草品种生产性能比较与优势品种筛选.草业学报,2014,23(4):39-48.
[10] Wallace J M,Curran W S,Mirsky S B,Ryan M R. Tolerance of interseeded annual ryegrass and red clover cover crops to residual herbicides in mid-atlantic corn cropping systems. Weed Technology,2017,31(5):641-650.
[11]刘爱荣,张远兵,张雪梅,何小丽,吴倩.空心莲子草水浸液对黑麦草和高羊茅种子发芽和幼苗生长的影响.草业学报,2007,16(5):96-101.
[12]许能祥,顾洪如,丁成龙,董臣飞,程云辉,张文洁,王兴刚.多花黑麦草耐盐性及其在盐土条件下饲用品质的研究.草业学报,2013,22(4):89-98.
[13]魏晓艳,梁丹妮,庞丁铭,兰剑. 14个多年生黑麦草品种幼苗期对盐胁迫的生理响应.安徽农业科学,2017,45(1):8-12.
[14]梁小红,艾非凡,钟天秀,韩烈保.多年生黑麦草对干旱-低温交叉适应的生理响应.草业学报,2016,25(1):163-170.
[15]冯鹏,孙力,申晓慧,姜成,李如来,李增杰,郑海燕,张华,郭伟,韩旭东,洪亚楠.应及富集能力的研究.草业学报,2016,25(1):153-162.
[16] Zhu J K. Plant salt tolerance. Trends in Plant Science,2001,6(2):66-71.
[17]薛延丰,石志琦,严少华,郑建初,常志州.利用生理生化参数评价水葫芦沼液浸种可行性初步研究.草业学报,2010,19(5):51-56.
[18]胡星云,孙志高,张党玉,孙文广,祝贺,任鹏.黄河口不同氮基质碱蓬种子萌发及幼苗生长对盐分及氮输入的响应.生态学报,2017,37(24):8499-8510.
[19]李兵兵,魏小红,徐严.麻花秦艽种子休眠机理及其破除方法.生态学报,2013,33(15):4631-4638.
[20]姜义宝,郑秋红,王成章,郭玉霞,李德锋.超干贮藏对菊苣种子活力与抗氧化性的影响.草业学报,2009,18(5):93-97.
[21]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.
[22] Zhang Z L,Qu W J,Li X F. Experimental Guide of Plant Physiology. 4th ed. Beijing:Higher Education Press,2009.(in Chinese)
[23] Wang A G,Luo G H. Quantitative relation between the reaction of hydroxylamine and superoxide anion radicals in plants. Plant Physiology Communications,1990,26(6):55-57.
[24] Jana S,Choudhuri M A. Glycolate metabolism of three submersed aquatic angiosperms during ageing. Aquatic Botany,1982,12:345-354.
[25] Xu X Y,Fan R,Zheng R,Li C M,Yu D Y. Proteomic analysis of seed germination under salt stress in soybeans. Journal of Zhejiang University Science B:Biomedicine&Biotechnology. 2011,12(7):507-517.
[26]王玉萍,董雯,张鑫,杨茜,张峰.水杨酸对盐胁迫下花椰菜种子萌发及幼苗生理特性的影响.草业学报,2012,21(1):213-219.
[27]杨景宁,王彦荣. Na Cl胁迫对四种荒漠植物种子萌发的影响.草业学报,2012,21(5):32-38.
[28] Li J S. Studies on the mechanism of regulations of G6PDH,c GMP,H2O2and Ca2+in plant adaptation to salt tolerance[D]. Lanzhou:Lanzhou University,2011.
[29]范燕,闵丹丹,郭正刚,冯葆昌,胡小文.多年生黑麦草种子萌发及苗期耐盐性的比较.草业科学,2017,34(4):724-734.
[30]樊瑞苹,周琴,周波,江海东.盐胁迫对高羊茅生长及抗氧化系统的影响.草业学报,2012,21(1):112-117.
[31] Li S W,Xue L G. The interaction between H2O2and NO,Ca2+,c GMP,and MAPKS during adventitious rooting in mung bean seedlings. In Vitro Cellular&Developmental Biology-Plant,2010,46(2):142-148.
[32]张巍巍,郑飞翔,王效科,冯兆忠,欧阳志云.臭氧对水稻根系活力、可溶性蛋白含量与抗氧化系统的影响.植物生态学报,2009,33(3):425-432.
[33]列淦文,叶龙华,薛立.臭氧胁迫对植物主要生理功能的影响.生态学报,2014,34(2):294-306.
[34] Kim Y H,Lim S,Han S H,Lee J C,Song W K,Bang W K,Kwon S Y,Lee H S,Kwak S S. Differential expression of 10 sweetpotato peroxidases in response to sulfur dioxide,ozone,and ultraviolet radiation. Plant Physiology and Biochemistry,2007,45(12):908-914.
[35] Szmidt-Jaworska A,Jaworski K,Kopcewicz J. Cyclic GMP stimulates flower induction of Pharbitis nil via its influence on c GMP regulated protein kinase. Plant Growth Regulation,2009,57(2):115-126.
[36] Suita K,Kiryu T,Sawada M,Mitsui M,Nakagawa M,Kanamaru K,Yamagata H. Cyclic GMP acts as a common regulator for the transcriptional activation of the flavonoid biosynthetic pathway in soybean. Planta,2009,229(2):403-413.
[37]樊怀福,郭世荣,焦彦生,张润花,李娟.外源一氧化氮对Na Cl胁迫下黄瓜幼苗生长、活性氧代谢和光合特性的影响.生态学报,2007,27(2):546-553.
[38]张国明,顾为,吴之正,史培军,王静爱.海冰水不同盐含量处理对棉花、小麦和玉米种子萌发影响.北京师范大学:自然科学版,2006,42(2):209-212.
[2] Irving H R,Gehring C. Molecular methods for the study of signal transduction in plants//Gehring C,ed. Cyclic Nucleotide Signaling in Plants.Totowa,NJ:Humana Press,2013,1016:1-11.
[3] Pagnussat G C,Lanteri M L,Lamattina L. Nitric oxide and cyclic GMP are messengers in the indole acetic acid-induced adventitious rooting process. Plant Physiology,2013,132(3):1241-1248.
[4] Meier S,Gehring C. Emerging roles in plant biotechnology for the second messenger c GMP-guanosine 3',5'-cyclic monophosphate. African Journal of Biotechnology,2006,5(19):1687-1692.
[5] Isner J C,Maathuis F J M. Measurement of cellular c GMP in plant cells and tissues using the endogenous fluorescent reporter Flinc G. Plant Journal,2011,65(2):329-334.
[6] Turek I,Gehring C. The plant natriuretic peptide receptor is a guanylyl cyclase and enables c GMP-dependent signaling. Plant Molecular Biology,2016,91(3):275-286.
[7] Durner J,Wendehenne D,Klessig D F. Defense gene induction in tobacco by nitric oxide,cyclic GMP,and cyclic ADP-ribose. Proceedings of the National Academy of Sciences of the United States of America,1998,95(17):10328-10333.
[8] Rubio F,Flores P,Navarro J M,Martínez V. Effects of Ca2+,K+and c GMP on Na+uptake in pepper plants. Plant Science,2003,165(5):1043-1049.
[9]刘春英,张学映,朱体超,陈光蓉,郑章云.不同黑麦草品种生产性能比较与优势品种筛选.草业学报,2014,23(4):39-48.
[10] Wallace J M,Curran W S,Mirsky S B,Ryan M R. Tolerance of interseeded annual ryegrass and red clover cover crops to residual herbicides in mid-atlantic corn cropping systems. Weed Technology,2017,31(5):641-650.
[11]刘爱荣,张远兵,张雪梅,何小丽,吴倩.空心莲子草水浸液对黑麦草和高羊茅种子发芽和幼苗生长的影响.草业学报,2007,16(5):96-101.
[12]许能祥,顾洪如,丁成龙,董臣飞,程云辉,张文洁,王兴刚.多花黑麦草耐盐性及其在盐土条件下饲用品质的研究.草业学报,2013,22(4):89-98.
[13]魏晓艳,梁丹妮,庞丁铭,兰剑. 14个多年生黑麦草品种幼苗期对盐胁迫的生理响应.安徽农业科学,2017,45(1):8-12.
[14]梁小红,艾非凡,钟天秀,韩烈保.多年生黑麦草对干旱-低温交叉适应的生理响应.草业学报,2016,25(1):163-170.
[15]冯鹏,孙力,申晓慧,姜成,李如来,李增杰,郑海燕,张华,郭伟,韩旭东,洪亚楠.应及富集能力的研究.草业学报,2016,25(1):153-162.
[16] Zhu J K. Plant salt tolerance. Trends in Plant Science,2001,6(2):66-71.
[17]薛延丰,石志琦,严少华,郑建初,常志州.利用生理生化参数评价水葫芦沼液浸种可行性初步研究.草业学报,2010,19(5):51-56.
[18]胡星云,孙志高,张党玉,孙文广,祝贺,任鹏.黄河口不同氮基质碱蓬种子萌发及幼苗生长对盐分及氮输入的响应.生态学报,2017,37(24):8499-8510.
[19]李兵兵,魏小红,徐严.麻花秦艽种子休眠机理及其破除方法.生态学报,2013,33(15):4631-4638.
[20]姜义宝,郑秋红,王成章,郭玉霞,李德锋.超干贮藏对菊苣种子活力与抗氧化性的影响.草业学报,2009,18(5):93-97.
[21]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.
[22] Zhang Z L,Qu W J,Li X F. Experimental Guide of Plant Physiology. 4th ed. Beijing:Higher Education Press,2009.(in Chinese)
[23] Wang A G,Luo G H. Quantitative relation between the reaction of hydroxylamine and superoxide anion radicals in plants. Plant Physiology Communications,1990,26(6):55-57.
[24] Jana S,Choudhuri M A. Glycolate metabolism of three submersed aquatic angiosperms during ageing. Aquatic Botany,1982,12:345-354.
[25] Xu X Y,Fan R,Zheng R,Li C M,Yu D Y. Proteomic analysis of seed germination under salt stress in soybeans. Journal of Zhejiang University Science B:Biomedicine&Biotechnology. 2011,12(7):507-517.
[26]王玉萍,董雯,张鑫,杨茜,张峰.水杨酸对盐胁迫下花椰菜种子萌发及幼苗生理特性的影响.草业学报,2012,21(1):213-219.
[27]杨景宁,王彦荣. Na Cl胁迫对四种荒漠植物种子萌发的影响.草业学报,2012,21(5):32-38.
[28] Li J S. Studies on the mechanism of regulations of G6PDH,c GMP,H2O2and Ca2+in plant adaptation to salt tolerance[D]. Lanzhou:Lanzhou University,2011.
[29]范燕,闵丹丹,郭正刚,冯葆昌,胡小文.多年生黑麦草种子萌发及苗期耐盐性的比较.草业科学,2017,34(4):724-734.
[30]樊瑞苹,周琴,周波,江海东.盐胁迫对高羊茅生长及抗氧化系统的影响.草业学报,2012,21(1):112-117.
[31] Li S W,Xue L G. The interaction between H2O2and NO,Ca2+,c GMP,and MAPKS during adventitious rooting in mung bean seedlings. In Vitro Cellular&Developmental Biology-Plant,2010,46(2):142-148.
[32]张巍巍,郑飞翔,王效科,冯兆忠,欧阳志云.臭氧对水稻根系活力、可溶性蛋白含量与抗氧化系统的影响.植物生态学报,2009,33(3):425-432.
[33]列淦文,叶龙华,薛立.臭氧胁迫对植物主要生理功能的影响.生态学报,2014,34(2):294-306.
[34] Kim Y H,Lim S,Han S H,Lee J C,Song W K,Bang W K,Kwon S Y,Lee H S,Kwak S S. Differential expression of 10 sweetpotato peroxidases in response to sulfur dioxide,ozone,and ultraviolet radiation. Plant Physiology and Biochemistry,2007,45(12):908-914.
[35] Szmidt-Jaworska A,Jaworski K,Kopcewicz J. Cyclic GMP stimulates flower induction of Pharbitis nil via its influence on c GMP regulated protein kinase. Plant Growth Regulation,2009,57(2):115-126.
[36] Suita K,Kiryu T,Sawada M,Mitsui M,Nakagawa M,Kanamaru K,Yamagata H. Cyclic GMP acts as a common regulator for the transcriptional activation of the flavonoid biosynthetic pathway in soybean. Planta,2009,229(2):403-413.
[37]樊怀福,郭世荣,焦彦生,张润花,李娟.外源一氧化氮对Na Cl胁迫下黄瓜幼苗生长、活性氧代谢和光合特性的影响.生态学报,2007,27(2):546-553.
[38]张国明,顾为,吴之正,史培军,王静爱.海冰水不同盐含量处理对棉花、小麦和玉米种子萌发影响.北京师范大学:自然科学版,2006,42(2):209-212.