PEG胁迫下两个大豆品种苗期的耐旱性与相关响应基因表达分析
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摘要
为揭示耐旱大豆材料对干旱胁迫的生理和分子反应,以耐旱型大豆品种‘吉育88’和‘杂交豆5号’为试验材料,研究中度水分胁迫(10%PEG6000)下9个耐旱生理指标变化和4个干旱胁迫响应基因(BADH1,GmDREBa, GmDREBb和GmERF3)的表达情况,明确大豆中相关干旱响应基因的表达与耐旱生理指标之间的相关性。结果表明,两种大豆材料在干旱胁迫下主要以提高可溶性糖、可溶性蛋白、淀粉含量以及保持较高的过氧化氢酶(CAT)活性来减少伤害。‘吉育88’和‘杂交豆5号’叶片中4个耐旱基因的表达模式基本一致,均为随胁迫时间的延长先升高后降低。相关性分析表明,‘杂交豆5号’BADH1基因在干旱胁迫9 h的相对表达量与CAT活性显著正相关;GmDREBa基因在干旱胁迫9 h的相对表达量与淀粉含量显著正相关;GmDREBb基因在干旱胁迫24 h的相对表达量与淀粉含量显著正相关;GmERF3基因在干旱胁迫24 h的相对表达量与脱落酸(ABA)含量显著正相关,而在干旱胁迫48 h的相对表达量与细胞分裂素(CTK)含量显著负相关。在考察大豆耐旱性时,这些基因的表达情况可以作为参考指标。
To reveal the physiological and molecular responses of drought-tolerant soybean materials to drought stress, the drought-to lerant soybean varieties 'Jiyu 88' and 'Hybrid bean 5' were used to study the changes of 9 physiological indexes of drought tolerance and the expression of 4 drought stress response genes(BADH1, GmDREBa, GmDREBb, and GmERF3) under moderate water stress(10% PEG6000). The correlation between the expression of drought response genes and physiological indexes of drought tolerance in soybean was clarifyed. The results showed that the two soybean materials mainly increased soluble sugar, soluble protein as well as starch content, and kept high catalase(CAT) activity to reduce injury under drought stress. The expression patterns of four drought-tolerant genes in leaves of 'Jiyu 88' and 'Hybrid bean 5' were basically the same, all of which increased at first and then decreased with the prolongation of stress time. The correlation analysis showed that there was a significant positive correlation between the relative expression of BADH1 gene and the activity of CAT in'Hybrid bean 5' under drought stress for 9 h. Under drought stress for 9 h, there existed a significant positive correlation between the relative expression of GmDREBa gene and starch content. A significant positive correlation was found between the relative expression of GmDREBb gene and starch content under drought stress for 24 h. However, there was a significant negative correlation between the relative expression and the content of cytokinin(CTK) under drought stress for 48 h. Based above results, the expression of these genes could be used as a reference index in the study of soybean drought tolerance.
To reveal the physiological and molecular responses of drought-tolerant soybean materials to drought stress, the drought-to lerant soybean varieties 'Jiyu 88' and 'Hybrid bean 5' were used to study the changes of 9 physiological indexes of drought tolerance and the expression of 4 drought stress response genes(BADH1, GmDREBa, GmDREBb, and GmERF3) under moderate water stress(10% PEG6000). The correlation between the expression of drought response genes and physiological indexes of drought tolerance in soybean was clarifyed. The results showed that the two soybean materials mainly increased soluble sugar, soluble protein as well as starch content, and kept high catalase(CAT) activity to reduce injury under drought stress. The expression patterns of four drought-tolerant genes in leaves of 'Jiyu 88' and 'Hybrid bean 5' were basically the same, all of which increased at first and then decreased with the prolongation of stress time. The correlation analysis showed that there was a significant positive correlation between the relative expression of BADH1 gene and the activity of CAT in'Hybrid bean 5' under drought stress for 9 h. Under drought stress for 9 h, there existed a significant positive correlation between the relative expression of GmDREBa gene and starch content. A significant positive correlation was found between the relative expression of GmDREBb gene and starch content under drought stress for 24 h. However, there was a significant negative correlation between the relative expression and the content of cytokinin(CTK) under drought stress for 48 h. Based above results, the expression of these genes could be used as a reference index in the study of soybean drought tolerance.
引文
An Y.Y.,Hao W.F.,Gong C.M.,Han R.L.,and Liang Z.S.,2010,Effects of drying and re-watering on the photosynthesis and active oxygen metabolism of Periplocasepium seedlings,Yingyong Shengtai Xuebao(Journal of Applied Ecology),21(12):3047-3055(安玉艳,郝文芳,龚春梅,韩蕊莲,梁宗锁,2010,干旱-复水处理对杠柳幼苗光合作用及活性氧代谢的影响,应用生态学报,21(12):3047-3055)
Barbosa H.S.,Arruda S.C.,Azevedo R.A.,and Arruda M.A.Z.,2012,New insights on proteomics of transgenic soybean seeds:evaluation of differential expressions of enzymes and proteins,Anal.Bioanal.Chem.,402(1):299-314
Costa M.A.,Pinheiro H.A.,Shimizu E.S.C.,Fonseca F.T.,Filho B.G.D.S.,Moraes F.K.C.,and Figueiredo D.M.D.,2010,Lipid peroxidation,chloroplastic pigments and antioxidant strategies in Carapa guianensis(Aubl.)subjected to waterdeficit and short-term rewetting,Trees(Berlin),24(2):275-283
Dong Z.,and Xie F.T.,1995,Effects of soil water stress on enzyme activity and membrane permeability in soybean,Dadou Kexue(Soybean Science),14(4):290-297(董钻,谢甫绨,1995,土壤水分胁迫对大豆体内酶活性和膜透性的影响,大豆科学,14(4):290-297)
Dubois M.,Gilles K.A.,and Hamilton J.K.,1951,A colorimetric method for the determination of sugars,Nature,168(4265):167
Gao X.Y.,Liu L.J.,Liu B.,Ma Y.L.,Wang R.Q.,and Dong S.K.,2016,Effect of drought stress simulated by PEG on antioxidant enzyme activities and antioxidant capacity in soybean,Dadou Kexue(Soybean Science),35(4):616-619,636(高鑫宇,刘丽君,刘博,马玉玲,王瑞奇,董守坤,2016,PEG模拟干旱对大豆抗氧化酶活性及抗氧化能力的影响,大豆科学,35(4):616-619,636)
Guo S.J.,Yang K.M.,Huo J.,Zhou Y.H.,Wang Y.P.,and Li G.Q.,2015,Influence of drought on leaf photosynthetic capacity and root growth of soybeans at grain filling stage,Yingyong Shengtai Xuebao(Journal of Applied Ecology),26(5):1419-1425(郭数进,杨凯敏,霍瑾,周永航,王燕平,李贵全,2015,干旱胁迫对大豆鼓粒期叶片光合能力和根系生长的影响,应用生态学报,26(5):1419-1425)
Li J.Y.,Zhou C.J.,Yang L.,Wu Y.K.,Du Z.Q.,Yang L.,and Tian Z.Y.,2010,Effect of water stress on endogenous hormone and protective enzymes in soybean seedling leaves,Dadou Kexue(Soybean Science),29(6):959-963(李建英,周长军,杨柳,吴耀坤,杜志强,杨丽,田中艳,2010,水分胁迫对大豆苗期叶片内源激素含量与保护酶活性的影响,大豆科学,29(6):959-963)
Li X.P.,Tian A.G.,Luo G.Z.,Gong Z.Z.,Zhang J.S.,and Chen S.Y.,2005,Soybean DRE-binding transcription factors that are responsive to abiotic stresses,Theor.Appl.Genet.,110(8):1355-1362
Li Y.N.,Li J.Y.,Wang G.N.,Dong M.,Liu W.Q.,and Ma F.M.,2007,Studies on mechanism of endogenous hormones in soybean seedling under water stress,Dadou Kexue(Soybean Science),26(4):627-629,636(李玉梅,李建英,王根林,董梅,刘文清,马凤鸣,2007,水分胁迫对大豆幼苗叶片内源激素的影响,大豆科学,26(4):627-629,636)
Li Y.P.,Ye W.,Wang M.,and Yan X.D.,2009,Climate change and drought:a risk assessment of crop-yield impacts,Climate Research,39:31-46
Marisha S.,Gupta S.K.,Baisakhi M.,Maurya V.K.,Farah D.,Afroz A.,and Vivek P.,2018,Proteomics unravel the regulating role of salicylic acid in soybean under yield limiting drought stress,Plant Physiol.Biochem.,130:529-541
Masia A.,Pitacco A.,Braggio L.,and Giulivo C.,1994,Hormonal responses to partial drying of the root system of Helianthus annuus,J.Exp.Bot.,45(1):69-76
Miura K.,Jin J.B.,Lee J.,Yoo C.Y.,Stirm V.,Miura T.,Ashworth E.N.,Bressan R.A.,Yun D.J.,and Hasegawa P.M.,2007,SIZ1-mediated sumoylation of ICE1 controls CBF3/DREB1A expression and freezing tolerance in Arabidopsis,Plant Cell,19(4):1403-1414
Qin D.,Zhao C.L.,Zheng C.Z.,and Wang P.W.,2015,Drought tolerance of transgenic soybean with BADH gene,Zhongguo Youliao Zuowu Xuebao(Chinese Journal of Oil Crops),37(6):752-758(秦迪,赵翠兰,郑成忠,王丕武,2015,转BADH基因大豆耐旱性分析,中国油料作物学报,37(6):752-758)
Singh A.,Singh P.K.,Singh R.,Pandit A.,Mahato A.K.,Gupta D.K.,Tyagi K.,Singh A.K.,Singh N.K.,and Sharma T.R.,2010,SNP haplotypes of the BADH1 gene and their association with aroma in rice(Oryza sativa L.),Mol.Breeding,26(2):325-338
Song L.,Prince S.,Valliyodan B.,Joshi T.,Santos J.V.M.D.,Wang J.,Lin L.,Wan J.R.,Wang Y.Q.,Xu D.,and Nguyen H.,2016,Genome-wide transcriptome analysis of soybean primary root under varying water-deficit conditions,BMCGenomics,17(1):57
Song Y.S.,Yin T.F.,Wang Y.Z.,Xue J.,and Liu L.J.,1987,Resistance of soybean to drought stress,Dadou Kexue(Soybean Science),(6):277-282(宋英淑,尹田夫,王以芝,薛津,刘丽君,1987,大豆对干旱胁迫的抗性效应,大豆科学,(6):277-282)
Tang W.,Sun J.,Liu J.,Liu F.,Yan J.,Gou X.,Lu B.R.,and Liu Y.,2014,RNAi-directed downregulation of betaine aldehyde dehydrogenase 1(OsBADH1)results in decreased stress tolerance and increased oxidative markers without affecting glycine betaine biosynthesis in rice(Oryza sativa),Plant Mol.Biol.,86(4-5):443-454
Wang F.,Chen H.W.,Li Q.T.,Wei W.,Li W.,Zhang W.K.,Ma B.,Bi Y.C.,Lai Y.C.,Liu X.L.,Man W.Q.,Zhang J.S.,and Chen S.Y.,2015,Gm WRKY27 interacts with Gm MYB174to reduce expression of GmNAC29 for stress tolerance in soybean plants,Plant J.,83(2):224-236
Wang W.W.,Lin H.,Tang X.F.,Wei L.,Dong X.Y.,Wu G.X.,and Liu L.J.,2014,Soybean drought resistance related genes expression characteristics under drought,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),12(5):903-908(王伟威,林浩,唐晓飞,魏崃,董兴月,吴广锡,刘丽君,2014,干旱胁迫下大豆相关基因的表达特性,分子植物育种,12(5):903-908)
Zhang G.,Chen M.,Li L.,Xu Z.,Chen X.,Guo J.,and Ma Y.,2009,Overexpression of the soybean GmERF3 gene,an AP2/ERF type transcription factor for increased tolerances to salt,drought,and diseases in transgenic tobacco,J.Exp.Bot.,60(13):3781-3796
Zhao Z.N.,and Zhao B.X.,2018,Physiological response and drought resistance evaluation of different soybean varieties to drought stress at germination stage,Ganhan Diqu Nongye Yanjiu(Agricultural Research in the Arid Areas),36(2):131-136(赵振宁,赵宝勰,2018,不同大豆品种在萌发期对干旱胁迫的生理响应及抗旱性评价,干旱地区农业研究,36(2):131-136)
Zhou Y.F.,Wang D.Q.,Lu Z.L.,Wang N.,Wang Y.T.,Li F.X.,Xu W.J.,and Huang R.D.,2014,Effects of drought stress on photosynthetic characteristics and endogenous hormone A-BA and CTK contents in green-stayed sorghum,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),47(4):655-663(周宇飞,王德权,陆樟镳,王娜,王艺陶,李丰先,许文娟,黄瑞冬,2014,干旱胁迫对持绿性高粱光合特性和内源激素ABA、CTK含量的影响,中国农业科学,47(4):655-663)
Barbosa H.S.,Arruda S.C.,Azevedo R.A.,and Arruda M.A.Z.,2012,New insights on proteomics of transgenic soybean seeds:evaluation of differential expressions of enzymes and proteins,Anal.Bioanal.Chem.,402(1):299-314
Costa M.A.,Pinheiro H.A.,Shimizu E.S.C.,Fonseca F.T.,Filho B.G.D.S.,Moraes F.K.C.,and Figueiredo D.M.D.,2010,Lipid peroxidation,chloroplastic pigments and antioxidant strategies in Carapa guianensis(Aubl.)subjected to waterdeficit and short-term rewetting,Trees(Berlin),24(2):275-283
Dong Z.,and Xie F.T.,1995,Effects of soil water stress on enzyme activity and membrane permeability in soybean,Dadou Kexue(Soybean Science),14(4):290-297(董钻,谢甫绨,1995,土壤水分胁迫对大豆体内酶活性和膜透性的影响,大豆科学,14(4):290-297)
Dubois M.,Gilles K.A.,and Hamilton J.K.,1951,A colorimetric method for the determination of sugars,Nature,168(4265):167
Gao X.Y.,Liu L.J.,Liu B.,Ma Y.L.,Wang R.Q.,and Dong S.K.,2016,Effect of drought stress simulated by PEG on antioxidant enzyme activities and antioxidant capacity in soybean,Dadou Kexue(Soybean Science),35(4):616-619,636(高鑫宇,刘丽君,刘博,马玉玲,王瑞奇,董守坤,2016,PEG模拟干旱对大豆抗氧化酶活性及抗氧化能力的影响,大豆科学,35(4):616-619,636)
Guo S.J.,Yang K.M.,Huo J.,Zhou Y.H.,Wang Y.P.,and Li G.Q.,2015,Influence of drought on leaf photosynthetic capacity and root growth of soybeans at grain filling stage,Yingyong Shengtai Xuebao(Journal of Applied Ecology),26(5):1419-1425(郭数进,杨凯敏,霍瑾,周永航,王燕平,李贵全,2015,干旱胁迫对大豆鼓粒期叶片光合能力和根系生长的影响,应用生态学报,26(5):1419-1425)
Li J.Y.,Zhou C.J.,Yang L.,Wu Y.K.,Du Z.Q.,Yang L.,and Tian Z.Y.,2010,Effect of water stress on endogenous hormone and protective enzymes in soybean seedling leaves,Dadou Kexue(Soybean Science),29(6):959-963(李建英,周长军,杨柳,吴耀坤,杜志强,杨丽,田中艳,2010,水分胁迫对大豆苗期叶片内源激素含量与保护酶活性的影响,大豆科学,29(6):959-963)
Li X.P.,Tian A.G.,Luo G.Z.,Gong Z.Z.,Zhang J.S.,and Chen S.Y.,2005,Soybean DRE-binding transcription factors that are responsive to abiotic stresses,Theor.Appl.Genet.,110(8):1355-1362
Li Y.N.,Li J.Y.,Wang G.N.,Dong M.,Liu W.Q.,and Ma F.M.,2007,Studies on mechanism of endogenous hormones in soybean seedling under water stress,Dadou Kexue(Soybean Science),26(4):627-629,636(李玉梅,李建英,王根林,董梅,刘文清,马凤鸣,2007,水分胁迫对大豆幼苗叶片内源激素的影响,大豆科学,26(4):627-629,636)
Li Y.P.,Ye W.,Wang M.,and Yan X.D.,2009,Climate change and drought:a risk assessment of crop-yield impacts,Climate Research,39:31-46
Marisha S.,Gupta S.K.,Baisakhi M.,Maurya V.K.,Farah D.,Afroz A.,and Vivek P.,2018,Proteomics unravel the regulating role of salicylic acid in soybean under yield limiting drought stress,Plant Physiol.Biochem.,130:529-541
Masia A.,Pitacco A.,Braggio L.,and Giulivo C.,1994,Hormonal responses to partial drying of the root system of Helianthus annuus,J.Exp.Bot.,45(1):69-76
Miura K.,Jin J.B.,Lee J.,Yoo C.Y.,Stirm V.,Miura T.,Ashworth E.N.,Bressan R.A.,Yun D.J.,and Hasegawa P.M.,2007,SIZ1-mediated sumoylation of ICE1 controls CBF3/DREB1A expression and freezing tolerance in Arabidopsis,Plant Cell,19(4):1403-1414
Qin D.,Zhao C.L.,Zheng C.Z.,and Wang P.W.,2015,Drought tolerance of transgenic soybean with BADH gene,Zhongguo Youliao Zuowu Xuebao(Chinese Journal of Oil Crops),37(6):752-758(秦迪,赵翠兰,郑成忠,王丕武,2015,转BADH基因大豆耐旱性分析,中国油料作物学报,37(6):752-758)
Singh A.,Singh P.K.,Singh R.,Pandit A.,Mahato A.K.,Gupta D.K.,Tyagi K.,Singh A.K.,Singh N.K.,and Sharma T.R.,2010,SNP haplotypes of the BADH1 gene and their association with aroma in rice(Oryza sativa L.),Mol.Breeding,26(2):325-338
Song L.,Prince S.,Valliyodan B.,Joshi T.,Santos J.V.M.D.,Wang J.,Lin L.,Wan J.R.,Wang Y.Q.,Xu D.,and Nguyen H.,2016,Genome-wide transcriptome analysis of soybean primary root under varying water-deficit conditions,BMCGenomics,17(1):57
Song Y.S.,Yin T.F.,Wang Y.Z.,Xue J.,and Liu L.J.,1987,Resistance of soybean to drought stress,Dadou Kexue(Soybean Science),(6):277-282(宋英淑,尹田夫,王以芝,薛津,刘丽君,1987,大豆对干旱胁迫的抗性效应,大豆科学,(6):277-282)
Tang W.,Sun J.,Liu J.,Liu F.,Yan J.,Gou X.,Lu B.R.,and Liu Y.,2014,RNAi-directed downregulation of betaine aldehyde dehydrogenase 1(OsBADH1)results in decreased stress tolerance and increased oxidative markers without affecting glycine betaine biosynthesis in rice(Oryza sativa),Plant Mol.Biol.,86(4-5):443-454
Wang F.,Chen H.W.,Li Q.T.,Wei W.,Li W.,Zhang W.K.,Ma B.,Bi Y.C.,Lai Y.C.,Liu X.L.,Man W.Q.,Zhang J.S.,and Chen S.Y.,2015,Gm WRKY27 interacts with Gm MYB174to reduce expression of GmNAC29 for stress tolerance in soybean plants,Plant J.,83(2):224-236
Wang W.W.,Lin H.,Tang X.F.,Wei L.,Dong X.Y.,Wu G.X.,and Liu L.J.,2014,Soybean drought resistance related genes expression characteristics under drought,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),12(5):903-908(王伟威,林浩,唐晓飞,魏崃,董兴月,吴广锡,刘丽君,2014,干旱胁迫下大豆相关基因的表达特性,分子植物育种,12(5):903-908)
Zhang G.,Chen M.,Li L.,Xu Z.,Chen X.,Guo J.,and Ma Y.,2009,Overexpression of the soybean GmERF3 gene,an AP2/ERF type transcription factor for increased tolerances to salt,drought,and diseases in transgenic tobacco,J.Exp.Bot.,60(13):3781-3796
Zhao Z.N.,and Zhao B.X.,2018,Physiological response and drought resistance evaluation of different soybean varieties to drought stress at germination stage,Ganhan Diqu Nongye Yanjiu(Agricultural Research in the Arid Areas),36(2):131-136(赵振宁,赵宝勰,2018,不同大豆品种在萌发期对干旱胁迫的生理响应及抗旱性评价,干旱地区农业研究,36(2):131-136)
Zhou Y.F.,Wang D.Q.,Lu Z.L.,Wang N.,Wang Y.T.,Li F.X.,Xu W.J.,and Huang R.D.,2014,Effects of drought stress on photosynthetic characteristics and endogenous hormone A-BA and CTK contents in green-stayed sorghum,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),47(4):655-663(周宇飞,王德权,陆樟镳,王娜,王艺陶,李丰先,许文娟,黄瑞冬,2014,干旱胁迫对持绿性高粱光合特性和内源激素ABA、CTK含量的影响,中国农业科学,47(4):655-663)