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作物学报 ›› 2013, Vol. 39 ›› Issue (12): 2135-2144.doi: 10.3724/SP.J.1006.2013.02135
闵东红1,**,薛飞洋1, 2,**,马亚男1, 2,陈明2,徐兆师2,李连城2,刁现民2,贾冠清2,马有志2
MIN Dong-Hong1,**,XUE Fei-Yang1,2,**,MA Ya-Nan1,2,CHEN Ming2,*,XU Zhao-Shi2,LI Lian-Cheng2,DIAO Xian-Min2,JIA Guan-Qing2,MA You-Zhi2
摘要：
PP2Cs (2C type protein phosphatases)是一种单体丝氨酸/苏氨酸蛋白磷酸酶，在真核生物中，PP2Cs在脱落酸(ABA)、茉莉酸(JA)、水杨酸(SA)等激素信号传导途径中起着重要的调控作用。本研究通过序列比对，从谷子基因组中筛选出80个PP2C候选基因，聚类分析将其分为12个亚族(A、B、C、D、E1、E2、F1、F2、G、H、I、J)。与拟南芥PP2C基因家族比对表明，A~I为2个物种共有的亚族，J亚族只存在于谷子基因组中，L亚族只存在于拟南芥中。将谷子A亚族的10个成员命名为SiPP2CA1-10。基因表达谱分析表明，A亚族基因不同程度受ABA、干旱、高盐、低温和低氮诱导表达，其中，SiPP2CA6、SiPP2CA8在5种处理下诱导表达量都高。对10个A亚族成员的启动子分析发现，在这些基因的启动子序列中含有多种参与逆境胁迫应答的顺式作用元件，其中，SiPP2CA5、SiPP2CA6、SiPP2CA7、SiPP2CA8的启动子中含有参与低氮胁迫响应的元件。进一步研究发现，SiPP2CA8主要在根部表达，且在低氮胁迫下一直有较高的表达水平。亚细胞定位结果显示SiPP2CA8定位在细胞膜、细胞质、细胞核中；双分子荧光互补试验(BiFC)结果表明，SiPP2CA8与一个ABA受体类似蛋白SiRCAR3(基因号Si018317m.g)在细胞膜、细胞质及细胞核上互作，表明SiPP2CA8在谷子中可能参与ABA信号传导过程。
| [1]Stern A, Privman E, Rasis M, Lavi S, Pupko T. Evolution of the metazoan protein phosphatase 2C superfamily. J Mol Evol, 2007, 64: 61–70[2]Cohen P. The structure and regijlation of protein phosphatases. Ann Rev Biochem, 1989, 58: 453–508[3]Schweighofer A, Hirt H, Meskiene I. Plant PP2C phosphatases: emerging functions in stress signaling. Trends Plant Sci, 2004, 9: 236–243[4]Meskiene I, Baudouin E, Schweighofer A, Liwosz A, Jonak C, Rodriguez P L, Jelinek H, Hirt H. Stress-induced protein phosphatase 2C is a negative regulator of a mitogen-activated protein kinase. J Biol Chem, 2003, 278: 18945–18952[5]Shi Y G. Serine/threonine phosphatases: mechanism through structure. Cell, 2009, 139: 468–484[6]Sheen J. Mutational analysis of protein phosphatase 2C involved in abscisic acid signal transduction in higher plants. Proc Natl Acad Sci USA, 1998, 95: 975–980[7]Schweighofer A, Kazanaviciute V, Scheikl E, Teige M, Doczi R, Hirt H, Schwanninger M, Kant M, Schuurink R, Mauch F, Buchala A, Cardinale F, Meskiene I. The PP2C-type phosphatase AP2C1, which negatively regulates MPK4 and MPK6, modulates innate immunity, jasmonic acid, and ethylene levels in Arabidopsis. Plant Cell, 2007, 19: 2213–2224[8]Nishimura N, Okamoto M, Narusaka M, Yasuda M, Nakashita H, Shinozaki K, Narusaka Y, Hirayama T. ABA hypersensitive germination2-1 causes the activation of both abscisic acid and salicylic acid responses in Arabidopsis. Plant Cell Physiol, 2009, 50: 2112–2122[9]Hirayama T, Shinozaki K. Perception and transduction of abscisic acid signals: keys to the function of the versatile plant hormone ABA. Trends Plant Sci, 2007, 12: 343–351[10]Meyer K, Leube M P, Grill E. A protein phosphatase 2C involved in ABA signal transduction in Arabidopsis thaliana. Science, 1994, 264: 1452–1455[11]Jeffrey Leung S M, and Jérôme Giraudat. The Arabidopsis ABSCISIC ACID-INSENSlTIVE2 (AB12) and ABI1 genes encode homologous protein phosphatases 2C involved in abscisic acid signal transduction. Plant Cell, 1997, 9: 759–771[12]Angela Saez N A, Miguel Gonzalez Guzman, Mary Paz Gonzalez-Garcia, Carlos Nicolas, Oscar Lorenzo, Pedro L Rodriguez. Gain-of-function and loss-of-function phenotypes of the protein phosphatase 2C HAB1 reveal its role as a negative regulator of abscisic acid signalling. Plant J, 2004, 37: 354–369[13]Yoshida T, Nishimura N, Kitahata N, Kuromori T, Ito T, Asami T, Shinozaki K, Hirayama T. ABA-hypersensitive germination3 encodes a protein phosphatase 2C (AtPP2CA) that strongly regulates abscisic acid signaling during germination among Arabidopsis protein phosphatase 2Cs. Plant Physiol, 2006, 140: 115–126[14]Nishimura N, Yoshida T, Kitahata N, Asami T, Shinozaki K, Hirayama T. ABA-Hypersensitive Germination1 encodes a protein phosphatase 2C, an essential component of abscisic acid signaling in Arabidopsis seed. Plant J, 2007, 50: 935–949[15]Reyes D, Rodriguez D, Gonzalez-Garcia M P, Lorenzo O, Nicolas G, Garcia-Martinez J L, Nicolas C. Overexpression of a protein phosphatase 2C from beech seeds in Arabidopsis shows phenotypes related to abscisic acid responses and gibberellin biosynthesis. Plant Physiol, 2006, 141: 1414–1424[16]Saavedra X, Modrego A, Rodriguez D, Gonzalez-Garcia M P, Sanz L, Nicolas G, Lorenzo O. The nuclear interactor PYL8/RCAR3 of Fagus sylvatica FsPP2C1 is a positive regulator of abscisic acid signaling in seeds and stress. Plant Physiol, 2010, 152: 133–150[17]Liu L, Hu X, Song J, Zong X, Li D, Li D. Over-expression of a Zea mays L. protein phosphatase 2C gene (ZmPP2C) in Arabidopsis thaliana decreases tolerance to salt and drought. J Plant Physiol, 2009, 166: 531–542[18]Liu X, Zhu Y, Zhai H, Cai H, Ji W, Luo X, Li J, Bai X. AtPP2CG1, a protein phosphatase 2C, positively regulates salt tolerance of Arabidopsis in abscisic acid-dependent manner. Biochem Biophys Res Commun, 2012, 422: 710–715[19]Jia H F, Lu D, Sun J H, Li C L, Xing Y, Qin L, Shen Y Y. Type 2C protein phosphatase ABI1 is a negative regulator of strawberry fruit ripening. J Exp Bot, 2013 (DOI:10.1093/jxb/ert1028)[20]Xue T T, Wang D, Zhang S Z, Ehlting J, Ni F, Jakab S, Zheng C C, Zhong Y. Genome-wide and expression analysis of protein phosphatase 2C in rice and Arabidopsis. BMC Genom, 2008, 9: 550[21]Bhaskara G B, Nguyen T T, Verslues P E. Unique drought resistance functions of the highly ABA-induced clade A protein phosphatase 2Cs. Plant Physiol, 2012, 160: 379–395[22]Wang Z M, Devos K M, Liu C J, Wang R Q, Gale M D. Construction of RFLP-based maps of foxtail millet, Setaria italica (L.) P. Beauv. Theor Appl Genet, 1998, 96: 31–36[23]Doust A N, Kellogg E A, Devos K M, Bennetzen J L. Foxtail millet: a sequence-driven grass model system. Plant Physiol, 2009, 149: 137–141[24]Bennetzen J L, Schmutz J, Wang H, Percifield R, Hawkins J, Pontaroli A C, Estep M, Feng L, Vaughn J N, Grimwood J, Jenkins J, Barry K, Lindquist E, Hellsten U, Deshpande S, Wang X W, Wu X M, Mitros T, Triplett J, Yang X H, Ye C Y, Mauro-Herrera M, Wang L, Li P, Sharma M, Sharma R, Ronald P C, Panaud O, Kellogg E A, Brutnell T P, Doust A N, Tuskan G A, Rokhsar D, Devos K M. Reference genome sequence of the model plant Setaria. Nat Biotechnol, 2012, 30: 555–561[25]Zhang G Y, Liu X, Quan Z W, Cheng S F, Xu X, Pan S K, Xie M, Zeng P, Yue Z, Wang W L, Tao Y, Bian C, Han C L, Xia Q J, Peng X H, Cao R, Yang X H, Zhan D L, Hu J C, Zhang Y X, Li H N, Li H, Li N, Wang J Y, Wang C C, Wang R Y, Guo T, Cai Y J, Liu C Z, Xiang H T, Shi Q X, Huang P, Chen Q C, Li Y R, Wang J, Zhao Z H, Wang J. Genome sequence of foxtail millet (Setaria italica) provides insights into grass evolution and biofuel potential. Nat Biotechnol, 2012, 30: 549–554[26]Yoo S D, Cho Y H, Sheen J. Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nat Prot, 2007, 2: 1565–1572[27]Loppes R, Radoux M. Abscisic acid biosynthesis and catabolism. Ann Rev Plant Biol, 2005, 56: 165–185[28]Lim C W, Kim J H, Baek W, Kim B S, Lee S C. Functional roles of the protein phosphatase 2C, AtAIP1, in abscisic acid signaling and sugar tolerance in Arabidopsis. Plant Sci, 2012, 187: 83–88[29]Ma Y, Szostkiewicz I, Korte A, Moes D, Yang Y, Christmann A, Grill E. Regulators of PP2C phosphatase activity function as abscisic acid sensors. Science, 2009, 324: 1064–1068 |
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