一个木薯Aux/IAA转录因子的克隆及调节活性氧功能分析
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摘要
木薯是一种重要的粮食作物并且能够作为原料用于生产绿色能源。探究木薯中具有重要调节作用的相关基因,对于木薯产量的稳定具有重要的意义。Aux/IAAs在植物生长发育和胁迫时能够发挥重要的调节作用,但关于其在木薯中的作用的研究还不清楚。本研究以木薯品种华南124 (South China 124, SC124)为材料,对一个MeIAA进行了克隆,研究发现MeIAA在根、茎、叶均有表达。其中表达量最高的部位为叶片,同时H2O2处理可以诱导MeIAA表达,且MeIAA蛋白定位于核。此外,该转录因子能够促进活性氧积累和增加丙二醛的含量。本研究为进一步研究Aux/IAA转录因子在植物功能及其活性氧积累的关系提供了理论依据。
Cassava is an important food crops and can be used as raw material for the production of green energy.It is of great significance for the stability of cassava yield to explore the genes involved in cassava which play an important role in regulating cassava production. Aux/IAAs play important regulatory roles in plant growth and under stress. However, the related research in cassava remains unclear. In this research, a MeIAA was cloned by using South China 124(SC124) as the material. It was found that the MeIAA was expressed in roots, stems and leaves, and the expression level in leaves was the highest. At the same time, H2O2 treatment could induce MeIAA expression, and MeIAA protein was located in the nucleus. In addition, the transcription factor could promote the accumulation of reactive oxygen species and increase the content of malondialdehyde. The research might provide a theoretical basis for further studies on the relationship between Aux/IAA transcription factors in plant function and reactive oxygen species accumulation.
Cassava is an important food crops and can be used as raw material for the production of green energy.It is of great significance for the stability of cassava yield to explore the genes involved in cassava which play an important role in regulating cassava production. Aux/IAAs play important regulatory roles in plant growth and under stress. However, the related research in cassava remains unclear. In this research, a MeIAA was cloned by using South China 124(SC124) as the material. It was found that the MeIAA was expressed in roots, stems and leaves, and the expression level in leaves was the highest. At the same time, H2O2 treatment could induce MeIAA expression, and MeIAA protein was located in the nucleus. In addition, the transcription factor could promote the accumulation of reactive oxygen species and increase the content of malondialdehyde. The research might provide a theoretical basis for further studies on the relationship between Aux/IAA transcription factors in plant function and reactive oxygen species accumulation.
引文
Balagopalan C.,2002,Cassava utilization in food,feed and industry,In:Hillocks R.J.,and Thresh J.M.(eds.),Cassava Biology,CABI Publishing,Cali,Colombia,pp.301-318
akir B.,Ozan K.,and Olcay A.C.,2013,Genome-wide analysis of Aux/IAA genes in Vitis vinifera:cloning and expression profiling of a grape Aux/IAA gene in response to phytohormone and abiotic stresses,Acta Physiol.Plant,35(2):365-377
Chen Q.Q.,2017,Functional study of two NAC transcription factor in Brassica napus regulating reactive oxygen species accumulation and cell death,Thesis for M.S.,Northwest A&FUniversity,Supervisor:Jiang Y.Q.,pp.17(陈芹芹,2017,油菜中两个NAC转录因子调控活性氧累积及细胞死亡的分子机制解析,硕士学位论文,西北农林科技大学,导师:江元清,pp.17)
Cooke T.J.,Poli D.B.,Sztein A.E.,and Cohen J.D.,2002,Evolutionary patterns in auxin action,Plant Mol.Biol.,49(3-4):319-338
De-Pinto G.L.,Locato V.,and De-Gara L.,2012,Redox regulation in plant programmed cell death,Plant Cell Environ.,35(2):234-244
Fang J.,Pu W.H.,and Zhang H.J.,2010,The development status of cassava industry at home and abroad,Zhongguo Nongxue Tongbao(Chinese Agricultural Science Bulletin),26(16):353-361(方佳,濮文辉,张慧坚,2010,国内外木薯产业发展近况,中国农学通报,26(16):353-361)
Gechev T.S.,Van B.F.,Stone J.M.,Denev I.,and Laloi C.,2006,Reactive oxygen species as signals that modulate plant stress responses and programmed cell death,Bioessays,28(11):1091-1101
Israr M.,Jewell A.,Kumar D.,and Sahi S.V.,2011,Interactive effects of lead,copper,nickel and zinc on growth,metal uptake and antioxidative metabolism of Sesbania drummondii J.,Hazard.Mater.,186(2-3):1520-1526
Jia X.J.,Dong L.H.,Ding C.B.,Li X.,and Yuan M.,2013,Effects of drought stress on reactive oxygen species and their scavenging system in Chlorophytum capense var.Medio-pictum leaf,Caoye Xuebao(Act Prataculturae Sinica),22(5):248-255(贾学静,董立花,丁春邦,李旭,袁明,2013,干旱胁迫对金心吊兰叶片活性氧及其清除系统的影响,草业学报,22(5):248-255)
Li X.L.,Wei Y.X.,Hu W.,Shi H.T.,and Liu G.Y.,2017,Analysis of expression,localization and transcriptional activation activity of MebZIP9 transcription factor in cassava,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(11):4540-4546(李小林,韦运谢,胡伟,施海涛,刘国银,2017,MebZIP9转录因子的表达定位及转录激活活性分析,分子植物育种,15(11):4540-4546)
Lin W.Q.,2007,Functional analyses of Aux/IAA family member IAA2 by reverse genetic approaches in Arabidopsis thaliana,Dissertation for Ph.D.,Zhejiang University,Supervisiors:Zhu M.Y.,and Wang J.H.,pp.53(林伟强,2007,拟南芥Aux/IAA家族基因IAA2的反向遗传学功能研究,博士学位论文,浙江大学,导师:朱睦元,王君晖,pp.53)
Ludwigmüller J.,2015,Bacteria and fungi controlling plant growth by manipulating auxin:balance between development and defense,J.Plant Physiol.,172:4-12
Luo S.,2015,Regulation of auxin signaling and root growth by Aux/IAA transcription factor Os IAA9 in arabidopsis,Thesis for M.S.,Northeast Normal University,Supervisor:Wang S.C.,pp.26(罗莎,2015,Aux/IAA家族转录因子Os IAA9对拟南芥生长素信号转导及根部生长发育的调控,硕士学位论文,东北师范大学,导师:王树才,pp.26)
Luo T.,2005,Identification and classification of cassava(Manihot esculenta Crantz)germplasm based on SSR(simple sequence repeat)markers,Dissertation for Ph.D.,Guangxi University,Supervisors:Li Y.R.,and Ye Y.P.,pp.1(罗霆,2005,木薯种质资源的SSR鉴定与分类,博士学位论文,广西大学,导师:李杨瑞,叶燕萍,pp.1)
Niu F.F.,2015,Dissecting the molecular mechanisms of canola NAC103 and arabidopsis WSK1 transcription factors in regulating ROS-dependent cell death and low potassium response,respectively,Thesis for M.S.,Northwest A&F University,Supervisor:Jiang Y.Q.,pp.29(牛芳芳,2015,油菜NAC103与拟南芥WSK1转录因子分别调控ROS依赖的细胞死亡与低钾应答的分子机制解析,硕士学位论文,西北农林科技大学,导师:江元清,pp.29)
Niu F.F.,Wang B.,Wu F.F.,Yan J.L.,Li L.,Wang C.,Wang Y.Q.,Yang B.,and Jiang Y.Q.,2014,Canola(Brassica napus L.)NAC103,transcription factor gene is a novel player inducing reactive oxygen species accumulation and cell death in plants,Biochem.Biophys.Res.Commun.,454(1):30-35
Petrov V.,Hille J.,Mueller-Roeber B.,and Gechev T.S.,2015,Ros-mediated abiotic stress-induced programmed cell death in plants,Front.Plant Sci.,6:69
Pogány M.,Rad U.V.,Grün S.,DongóA.,Pintye A.,Simoneau P.,Bahnweg G.,Kiss L.,Barna B.,and Durner J.,2009,Dual roles of reactive oxygen species and nadph oxidase rbohd in an arabidopsis-Alternaria pathosystem,Plant Physiol.,151(3):1459-1475
Shi H.T.,Jiang C.,Ye T.T.,Tan D.X.,Reiter R.J.,Zhang H.,Liu R.Y.,and Chan Z.L.,2015,Comparative physiological,metabolomic,and transcriptomic analyses reveal mechanisms of improved abiotic stress resistance in bermudagrass(Cynodon dactylon(L).Pers.)by exogenous melatonin,J.Exp.Bot.,66(3):681-694
Su J.,Guo T.W.,and Ji K.S.,2015,Cloning and premilinary analysis of Aux/IAA family genes from Pinus massoniana,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),13(8):1822-1830(苏江,郭天玮,季孔庶,2015,马尾松Aux/IAA家族相关基因克隆及初步分析,分子植物育种,13(8):1822-1830)
Wang F.Q.,Tian Y.H.,Li M.J.,Yang J.F.,Zhang B.,Lin W.X.,Chen X.J.,and Zhang Z.Y.,2013,Molecular cloning and expression analysis of an Aux/IAA gene(RgIAA1)from Rehmannia glutinosa,Zhongguo Zhongyao Zazhi(China Journal of Chinese Materia Medica),38(23):4033-4039(王丰青,田云鹤,李明杰,杨金凤,张宝,林文雄,陈新建,张重义,2013,地黄Aux/IAA家族基因RgIAA1的克隆和表达分析,中国中药杂志,38(23):4033-4039)
Wang Y.Q.,He J.Z.,Zhao X.L.,Zhang H.Y.,Deng F.,Jia X.Y.,and Liu C.,2009,Comparison of salt-tolerance and droughttolerance of two desert plants(Karelinia caspica and Suaeda turkestanica litw.)in Tarim,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),28(6):1128-1134(王彦芹,贺江舟,赵小亮,张海燕,邓芳,贾晓宇,刘陈,2009,塔里木盆地荒漠植物花花柴和碱蓬耐盐,耐旱性比较,基因组学与应用生物学,28(6):1128-1134)
Wei Y.X.,Liu W.,Hu W.,Liu G.Y.,Wu C.J.,Liu W.,Zeng H.Q.,He C.Z.,and Shi H.T.,2017,Genome-wide analysis of autophagy-related genes in banana highlights MaATG8s,in cell death and autophagy in immune response to Fusarium wilt,Plant Cell Rep.,36(8):1237-1250
Worley C.K.,Zenser N.,Ramos J.,Rouse D.,Leyser O.,Theologis A.,and Callis J.,2010,Degradation of Aux/IAA proteins is essential for normal auxin signalling,Plant J.,21(6):553-562
Yang D.L.,Yang Y.,and He Z.,2013,Roles of plant hormones and their interplay in rice immunity,Mol.Plant,6(3):675-685
akir B.,Ozan K.,and Olcay A.C.,2013,Genome-wide analysis of Aux/IAA genes in Vitis vinifera:cloning and expression profiling of a grape Aux/IAA gene in response to phytohormone and abiotic stresses,Acta Physiol.Plant,35(2):365-377
Chen Q.Q.,2017,Functional study of two NAC transcription factor in Brassica napus regulating reactive oxygen species accumulation and cell death,Thesis for M.S.,Northwest A&FUniversity,Supervisor:Jiang Y.Q.,pp.17(陈芹芹,2017,油菜中两个NAC转录因子调控活性氧累积及细胞死亡的分子机制解析,硕士学位论文,西北农林科技大学,导师:江元清,pp.17)
Cooke T.J.,Poli D.B.,Sztein A.E.,and Cohen J.D.,2002,Evolutionary patterns in auxin action,Plant Mol.Biol.,49(3-4):319-338
De-Pinto G.L.,Locato V.,and De-Gara L.,2012,Redox regulation in plant programmed cell death,Plant Cell Environ.,35(2):234-244
Fang J.,Pu W.H.,and Zhang H.J.,2010,The development status of cassava industry at home and abroad,Zhongguo Nongxue Tongbao(Chinese Agricultural Science Bulletin),26(16):353-361(方佳,濮文辉,张慧坚,2010,国内外木薯产业发展近况,中国农学通报,26(16):353-361)
Gechev T.S.,Van B.F.,Stone J.M.,Denev I.,and Laloi C.,2006,Reactive oxygen species as signals that modulate plant stress responses and programmed cell death,Bioessays,28(11):1091-1101
Israr M.,Jewell A.,Kumar D.,and Sahi S.V.,2011,Interactive effects of lead,copper,nickel and zinc on growth,metal uptake and antioxidative metabolism of Sesbania drummondii J.,Hazard.Mater.,186(2-3):1520-1526
Jia X.J.,Dong L.H.,Ding C.B.,Li X.,and Yuan M.,2013,Effects of drought stress on reactive oxygen species and their scavenging system in Chlorophytum capense var.Medio-pictum leaf,Caoye Xuebao(Act Prataculturae Sinica),22(5):248-255(贾学静,董立花,丁春邦,李旭,袁明,2013,干旱胁迫对金心吊兰叶片活性氧及其清除系统的影响,草业学报,22(5):248-255)
Li X.L.,Wei Y.X.,Hu W.,Shi H.T.,and Liu G.Y.,2017,Analysis of expression,localization and transcriptional activation activity of MebZIP9 transcription factor in cassava,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(11):4540-4546(李小林,韦运谢,胡伟,施海涛,刘国银,2017,MebZIP9转录因子的表达定位及转录激活活性分析,分子植物育种,15(11):4540-4546)
Lin W.Q.,2007,Functional analyses of Aux/IAA family member IAA2 by reverse genetic approaches in Arabidopsis thaliana,Dissertation for Ph.D.,Zhejiang University,Supervisiors:Zhu M.Y.,and Wang J.H.,pp.53(林伟强,2007,拟南芥Aux/IAA家族基因IAA2的反向遗传学功能研究,博士学位论文,浙江大学,导师:朱睦元,王君晖,pp.53)
Ludwigmüller J.,2015,Bacteria and fungi controlling plant growth by manipulating auxin:balance between development and defense,J.Plant Physiol.,172:4-12
Luo S.,2015,Regulation of auxin signaling and root growth by Aux/IAA transcription factor Os IAA9 in arabidopsis,Thesis for M.S.,Northeast Normal University,Supervisor:Wang S.C.,pp.26(罗莎,2015,Aux/IAA家族转录因子Os IAA9对拟南芥生长素信号转导及根部生长发育的调控,硕士学位论文,东北师范大学,导师:王树才,pp.26)
Luo T.,2005,Identification and classification of cassava(Manihot esculenta Crantz)germplasm based on SSR(simple sequence repeat)markers,Dissertation for Ph.D.,Guangxi University,Supervisors:Li Y.R.,and Ye Y.P.,pp.1(罗霆,2005,木薯种质资源的SSR鉴定与分类,博士学位论文,广西大学,导师:李杨瑞,叶燕萍,pp.1)
Niu F.F.,2015,Dissecting the molecular mechanisms of canola NAC103 and arabidopsis WSK1 transcription factors in regulating ROS-dependent cell death and low potassium response,respectively,Thesis for M.S.,Northwest A&F University,Supervisor:Jiang Y.Q.,pp.29(牛芳芳,2015,油菜NAC103与拟南芥WSK1转录因子分别调控ROS依赖的细胞死亡与低钾应答的分子机制解析,硕士学位论文,西北农林科技大学,导师:江元清,pp.29)
Niu F.F.,Wang B.,Wu F.F.,Yan J.L.,Li L.,Wang C.,Wang Y.Q.,Yang B.,and Jiang Y.Q.,2014,Canola(Brassica napus L.)NAC103,transcription factor gene is a novel player inducing reactive oxygen species accumulation and cell death in plants,Biochem.Biophys.Res.Commun.,454(1):30-35
Petrov V.,Hille J.,Mueller-Roeber B.,and Gechev T.S.,2015,Ros-mediated abiotic stress-induced programmed cell death in plants,Front.Plant Sci.,6:69
Pogány M.,Rad U.V.,Grün S.,DongóA.,Pintye A.,Simoneau P.,Bahnweg G.,Kiss L.,Barna B.,and Durner J.,2009,Dual roles of reactive oxygen species and nadph oxidase rbohd in an arabidopsis-Alternaria pathosystem,Plant Physiol.,151(3):1459-1475
Shi H.T.,Jiang C.,Ye T.T.,Tan D.X.,Reiter R.J.,Zhang H.,Liu R.Y.,and Chan Z.L.,2015,Comparative physiological,metabolomic,and transcriptomic analyses reveal mechanisms of improved abiotic stress resistance in bermudagrass(Cynodon dactylon(L).Pers.)by exogenous melatonin,J.Exp.Bot.,66(3):681-694
Su J.,Guo T.W.,and Ji K.S.,2015,Cloning and premilinary analysis of Aux/IAA family genes from Pinus massoniana,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),13(8):1822-1830(苏江,郭天玮,季孔庶,2015,马尾松Aux/IAA家族相关基因克隆及初步分析,分子植物育种,13(8):1822-1830)
Wang F.Q.,Tian Y.H.,Li M.J.,Yang J.F.,Zhang B.,Lin W.X.,Chen X.J.,and Zhang Z.Y.,2013,Molecular cloning and expression analysis of an Aux/IAA gene(RgIAA1)from Rehmannia glutinosa,Zhongguo Zhongyao Zazhi(China Journal of Chinese Materia Medica),38(23):4033-4039(王丰青,田云鹤,李明杰,杨金凤,张宝,林文雄,陈新建,张重义,2013,地黄Aux/IAA家族基因RgIAA1的克隆和表达分析,中国中药杂志,38(23):4033-4039)
Wang Y.Q.,He J.Z.,Zhao X.L.,Zhang H.Y.,Deng F.,Jia X.Y.,and Liu C.,2009,Comparison of salt-tolerance and droughttolerance of two desert plants(Karelinia caspica and Suaeda turkestanica litw.)in Tarim,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),28(6):1128-1134(王彦芹,贺江舟,赵小亮,张海燕,邓芳,贾晓宇,刘陈,2009,塔里木盆地荒漠植物花花柴和碱蓬耐盐,耐旱性比较,基因组学与应用生物学,28(6):1128-1134)
Wei Y.X.,Liu W.,Hu W.,Liu G.Y.,Wu C.J.,Liu W.,Zeng H.Q.,He C.Z.,and Shi H.T.,2017,Genome-wide analysis of autophagy-related genes in banana highlights MaATG8s,in cell death and autophagy in immune response to Fusarium wilt,Plant Cell Rep.,36(8):1237-1250
Worley C.K.,Zenser N.,Ramos J.,Rouse D.,Leyser O.,Theologis A.,and Callis J.,2010,Degradation of Aux/IAA proteins is essential for normal auxin signalling,Plant J.,21(6):553-562
Yang D.L.,Yang Y.,and He Z.,2013,Roles of plant hormones and their interplay in rice immunity,Mol.Plant,6(3):675-685