拟南芥蛋白磷酸酶PP2C31的盐胁迫响应功能研究
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摘要
蛋白磷酸酶2C(protein phosphatase 2C,PP2Cs)家族是植物细胞去除磷酸化蛋白中磷酸基团的重要蛋白磷酸酶。拟南芥含有80多个编码PP2Cs的基因,然而大部分成员在植物抗逆反应中的功能仍有待研究。本文发现拟南芥PP2C31(At2g40860)基因的表达受高盐抑制,其T-DNA插入突变体(pp2c31-1突变体和pp2c31-2突变体)表现出对高盐胁迫的耐受性且Na+含量较低。实时定量PCR(quantitative real time-PCR,qRT-PCR)分析发现,PP2C31基因在植物体各组织均有表达;亚细胞定位结果显示PP2C31蛋白定位于细胞质和细胞核中。利用脱落酸合成抑制剂和外源脱落酸处理发现,突变体植株的高盐耐受性和PP2C31基因的表达均不受脱落酸的影响。研究结果表明:PP2C31蛋白是拟南芥响应高盐胁迫的一个非脱落酸依赖的负调控组分。
The protein phosphatase 2C(PP2Cs)family plays a critical role in the dephosphorylation of phosphorylated proteins in plant cells.The Arabidopsis thaliana genome contains over 80 genes encoding PP2 Cs,but the roles of most members in plant stress responses need further study.This paper reports that an Arabidopsis PP2 Cmember,PP2C31(At2g40860),functions in response to salt stress.The expression of PP2C31 gene was inhibited by salt stress,and its T-DNA insertion mutant lines(pp2c31-1 and pp2c31-2 mutant)exhibited a NaCl-resistant growth phenotype.Additionally,the Na+content contained in these mutants is less than that in the wild-type plants The quantitative real time-PCR(qRT-PCR)analysis showed that PP2C31 gene was expressed widely in Arabidopsis,and its encoding protein,detected by GFP labelling,is localized in the cytoplasm and nucleus.The NaCl resistance in pp2c31 mutant plants was little affected by the abscisic acid(ABA)synthesis inhibitor,and the expression level of PP2C31 gene was insensitive to exogenous ABA.These results demonstrate that PP2C31 protein is a negative component of ABA-independent signalling in response to salt stress in Arabidopsis.
The protein phosphatase 2C(PP2Cs)family plays a critical role in the dephosphorylation of phosphorylated proteins in plant cells.The Arabidopsis thaliana genome contains over 80 genes encoding PP2 Cs,but the roles of most members in plant stress responses need further study.This paper reports that an Arabidopsis PP2 Cmember,PP2C31(At2g40860),functions in response to salt stress.The expression of PP2C31 gene was inhibited by salt stress,and its T-DNA insertion mutant lines(pp2c31-1 and pp2c31-2 mutant)exhibited a NaCl-resistant growth phenotype.Additionally,the Na+content contained in these mutants is less than that in the wild-type plants The quantitative real time-PCR(qRT-PCR)analysis showed that PP2C31 gene was expressed widely in Arabidopsis,and its encoding protein,detected by GFP labelling,is localized in the cytoplasm and nucleus.The NaCl resistance in pp2c31 mutant plants was little affected by the abscisic acid(ABA)synthesis inhibitor,and the expression level of PP2C31 gene was insensitive to exogenous ABA.These results demonstrate that PP2C31 protein is a negative component of ABA-independent signalling in response to salt stress in Arabidopsis.
引文
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[11]MANABE Y,BRESSAN R A,WANG T,et al.The Arabidopsis kinase-associated protein phosphatase regulates adaptation to Na+stress[J].Plant Physiology,2008,146(2):612-622.
[12]COHEN P T.Novel protein serine/threonine phosphatases:variety is the spice of life[J].Trends in Biochemical Sciences,1997,22(7):245-251.
[13]XIONG L,SCHUMAKER K S,ZHU J K.Cell signaling during cold,drought,and salt stress[J].The Plant Cell,2002,14:S165-S183.
[14]SCHWEIGHOFER A,HIRT H,MESKIENE I.Plant PP2Cphosphatases:emerging functions in stress signaling[J].Trends in Plant Science,2004,9(5):236-243.
[15]XUE T,WANG D,ZHANG S,et al.Genome-wide and expression analysis of protein phosphatase 2Cin rice and Arabidopsis[J].BMC Genomics,2008,9:550.
[16]PARK S Y,FUNG P,NISHIMURA N,et al.Abscisic acid inhibits type 2Cprotein phosphatases via the PYR/PYL family of START proteins[J].Science,2009,324(5930):1068-1071.
[17]MA Y,SZOSTKIEWICZ I,KORTE A,et al.Regulators of PP2Cphosphatase activity function as abscisic acid sensors[J].Science,2009,324(5930):1064-1068.
[18]KRZYWIN′SKA E,KULIK A,BUCHOLC M,et al.Protein phosphatase type 2C PP2CA together with ABI1inhibits SnRK2.4activity and regulates plant responses to salinity[J].Plant Signaling and Behavior,2016,11(12):e1253647.
[19]KRZYWIN′SKA E,BUCHOLC M,KULIK A,et al.Phosphatase ABI1and okadaic acid-sensitive phosphoprotein phosphatases inhibit salt stress-activated SnRK2.4kinase[J].BMC Plant Biology,2016,16(1):136.
[2]ZHU J K.Plant salt tolerance[J].Trends in Plant Science,2001,6:66-71.
[3]ZHANG J L,SHI H.Physiological and molecular mechanisms of plant salt tolerance[J].Photosynthesis Research,2013,115(1):1-22.
[4]ZHU J K.Salt and drought stress signal transduction in plants[J].Annual Review of Plant Biology,2002,53:247-273.
[5]SHABALA S,SHABALA L.Ion transport and osmotic adjustment in plants and bacteria[J].Biomolecular Concepts,2011,2:407-419.
[6]DEINLEIN U,STEPHAN A B,HORIE T,et al.Plant salt-tolerance mechanisms[J].Trends in Plant Science,2014,19(6):371-379.
[7]SHI H,ISHITANI M,KIM C,et al.The Arabidopsis thaliana salt tolerance gene SOS1encodes a putative Na+/H+antiporter[J].Proceedings of the National Academy of Sciences of the United States of America,2000,97(12):6896-6901.
[8]LEE S C,LUAN S.ABA signal transduction at the crossroad of biotic and abiotic stress responses[J].Plant,Cell and Environment,2012,35(1):53-60.
[9]MESKIENE I,BAUDOUIN E,SCHWEIGHOFER A,et al.Stress-induced protein phosphatase 2Cis a negative regulator of a mitogen-activated protein kinase[J].Journal of Biological Chemistry,2003,278(21):18945-18952.
[10]LAMMERS T,LAVI S.Role of type 2Cprotein phosphatases in growth regulation and in cellular stress signaling[J].Critical Reviews in Biochemistry and Molecular Biology,2007,42(6):437-461.
[11]MANABE Y,BRESSAN R A,WANG T,et al.The Arabidopsis kinase-associated protein phosphatase regulates adaptation to Na+stress[J].Plant Physiology,2008,146(2):612-622.
[12]COHEN P T.Novel protein serine/threonine phosphatases:variety is the spice of life[J].Trends in Biochemical Sciences,1997,22(7):245-251.
[13]XIONG L,SCHUMAKER K S,ZHU J K.Cell signaling during cold,drought,and salt stress[J].The Plant Cell,2002,14:S165-S183.
[14]SCHWEIGHOFER A,HIRT H,MESKIENE I.Plant PP2Cphosphatases:emerging functions in stress signaling[J].Trends in Plant Science,2004,9(5):236-243.
[15]XUE T,WANG D,ZHANG S,et al.Genome-wide and expression analysis of protein phosphatase 2Cin rice and Arabidopsis[J].BMC Genomics,2008,9:550.
[16]PARK S Y,FUNG P,NISHIMURA N,et al.Abscisic acid inhibits type 2Cprotein phosphatases via the PYR/PYL family of START proteins[J].Science,2009,324(5930):1068-1071.
[17]MA Y,SZOSTKIEWICZ I,KORTE A,et al.Regulators of PP2Cphosphatase activity function as abscisic acid sensors[J].Science,2009,324(5930):1064-1068.
[18]KRZYWIN′SKA E,KULIK A,BUCHOLC M,et al.Protein phosphatase type 2C PP2CA together with ABI1inhibits SnRK2.4activity and regulates plant responses to salinity[J].Plant Signaling and Behavior,2016,11(12):e1253647.
[19]KRZYWIN′SKA E,BUCHOLC M,KULIK A,et al.Phosphatase ABI1and okadaic acid-sensitive phosphoprotein phosphatases inhibit salt stress-activated SnRK2.4kinase[J].BMC Plant Biology,2016,16(1):136.