盐胁迫下的杜梨PbPYL4基因克隆及其与PbNCED2基因表达分析Molecular cloning of PbPYL4 gene and expression analysis of PbPYL4 and PbNCED2 in Pyrus betulaefolia under salt stress
王宏;蔺经;李晓刚;王中华;常有宏;
摘要(Abstract):
【目的】分离和克隆杜梨ABA受体蛋白基因Pb PYL4,了解其在杜梨响应盐胁迫的表达情况。【方法】以杜梨(Pyrus betulaefolia Bunge)无性系组培苗为试验材料,使用RT-PCR的方法从杜梨中克隆到ABA受体基因,命名为PbPYL4,并研究该基因盐胁迫处理下的表达模式,同时检测ABA合成基因Pb NCED2在盐胁迫下的表达模式。【结果】PbPYL4的c DNA长度为682 bp,其中开放阅读框(ORF)621 bp,编码207个氨基酸的蛋白。系统发生分析结果表明,PbPYL4与植物中已经发现的拟南芥ABA受体蛋白PYL4/RCAR10亲缘关系较近。荧光定量RT-PCR表达分析结果表明,根,盐胁迫12 h内,Pb PYL4基因被诱导表达,12 h达到第1个小高峰,随后24 h表达下降,48 h表达又上升,72 h达到第2个峰值;茎,盐胁迫48 h后,Pb PYL4基因被诱导表达;叶片,处理12 h后,Pb PYL4基因被诱导表达,72 h表达略微下降。在同样组织中,Pb NCED2表现出与Pb PYL4相似的表达时序。【结论】Pb PYL4基因可能在杜梨响应盐胁迫的过程中起了重要的作用。
关键词(KeyWords): 杜梨;ABA受体PbPYL4;基因克隆;表达分析
基金项目(Foundation): 国家自然科学基金(31300469);; 江苏省农业科技自主创新[CX(13)5105,CX(14)5016];; 江苏省自然科学基金(BK2014021063)
作者(Author): 王宏;蔺经;李晓刚;王中华;常有宏;
Email:
DOI: 10.13925/j.cnki.gsxb.20140138
参考文献(References):
- [1]CHAVES M M,FLEXAS J,PINHEIRO C.Photosynthesis underdrought and salt stress:regulation mechanisms from whole plantto cell[J].Annals of Botany,2009,103:551-560.
- [2]BI Ying-dong,LIU Qing-xing,GUO Chang-hong,GUO Dong-lin.Advances of the ABA and the salt stress tolerance mecha-nismsin plant signal transduction pathways[J].Chinese Agricultur-al Science Bulletin,2013,29(9):167-171.毕影东,刘清醒,郭长虹,郭东林.ABA与植物耐盐信号转导途径的研究进展[J].中国农学通报,2013,29(9):167-171.
- [3]MUNNS R,JAMES R A,LAUCHLI A.Approaches to increasingthe salt tolerance of wheat and other cereals[J].Journal of Experi-mental Botany,2006,57:1025-1043.
- [4]CHAVES M M,MAROCO J P,PEREIRA J S.Understandingplant responses to drough-from genes to the whole plant[J].Func-tional Plant Biology,2003,30:239-264.
- [5]KIM T H,BOHMER M,HU H,NISHIMURA N,SCHROEDERJ I.Guard cell signal transduction network:advances in under-standing abscisic acid,CO2,and Ca2+signaling[J].Annual Re-view of Plant Biology,2010,61:561-591.
- [6]CULTER S R,RODRIGUEZ P L,FINKELSTEIN R R,ABARAMSS R.Abscisic acid:emergence of a core signaling network[J].AnnualReview of Plant Biology,2010,61:651-679.
- [7]PARK SY,FUNG P,NISHIMURA N,JENSEN D R,FUJII H,ZHAO Y,LUMBA S,SANTIAGO J,RODRIGUES A,CHOW TF,ALFRED S E,BONETTA D,FINKELSTEIN R,PROVARTN J,DESVEAUX D,RODRIGUEZ P L,MCCOURT P,ZHU JK,SCHROEDER J I,VOLKMAN B F,CUTLER S R.Abscisicacid inhibits type 2C protein phosphatases via the PYR/PYL fami-ly of START proteins[J].Science(New York,NY),2009,324:1068-1071.
- [8]HU Shuai,WANG Fang-zhan,LIU Zhen-ning,LIU Ya-pei,YU Xiao-lin.ABA signaling mediated by PYR/PYL/RCAR inplants[J].Hereditas(Beijing),2012,34(5):560-572.胡帅,王芳展,刘振宁,刘亚培,余小林.PYR/PYL/RCAR蛋白介导植物ABA的信号转导[J].遗传,2012,34(5):560-572.
- [9]YI Wen-kai,WANG Jia,YANG Hui,TIAN Yun,LU Xiang-yang.Plant ABA receptor and Signal transduction pathways[J].Chinese Bulletin of Botany,2012,47(5):515-524.易文凯,王佳,杨辉,田云,卢向阳.植物ABA受体及其介导的信号转导通路[J].植物学报,2012,47(5):515-524.
- [10]FUJII H,CHINNUSAMY V,RODRIGUES A,RUBIO S,AN-TONI R,PARK S Y,CULTER S R,SHEEN J,RODRIGUEZ PL,ZHU J K.In vitro reconstitution of an abscisic acid signallingpathway[J].Nature,2009,462:660-664.
- [11]MELCHER K,NG L M,ZHOU X E,SOON F F,XU Y,SUI-NO-POWELL K M,PARK S Y,WEINER J J,FUJII H,CHIN-NUSAMY V,KOVACH A,LI J,WANG Y,LI J,PETERSONF C,JENSEN D R,YONG E L,VOLKMAN B F,CUTLER SR,ZHU J K,XU H E.A gate-latch-lock mechanism for hor-mone signalling by abscisic acid receptors[J].Nature,2009,462:602-608.
- [12]MIYAZONO K,MIYAKAWA T,SAWANO Y,KUBOTA K,KANG H J,ASANO A,MIYAUCHI Y,TAKAHASHI M,ZHIY,FUJITA Y,YOSHIDA T,KODAIRA K S,YAMGUCHI-SHINOZAKI K,TANOKURA M.Structural basis of abscisic acidsignalling[J].Nature,2009,462:609-614.
- [13]NISHIMURA N,HITOMI K,ARVAI AS,RAMBO R P,HITO-MI C,CUTLER S R,SCHROEDER J I,GETZOFF E D.Struc-tural mechanism of abscisic acid binding and signaling by dimer-ic PYR1[J].Science(New York,NY),2009,326:1373-1379.
- [14]SANTIGO J,DUPEUX F,ROUND A,ANTONI R,PARK S Y,JAMIN M,CULTER S R,RODRIGUEZ P L,MARQUEZ J A.The abscisic acid receptor PYR1 in complex with abscisic acid[J].Nature,2009,462:665-668.
- [15]UMEZAWA T,SUGIYAMA N,MIZOGUCHI M,HAYASHI S,MYOUGA F,YAMAGUCHI-SHINOZAKI K,ISHIHAMA Y,HIRAYAMA T,SHINOZAKI K.Type 2C protein phosphatasesdirectly regulate abscisic acid-activated protein kinases in Arabi-dopsis[J].Proceedings of the National Academy of Sciences of theUnited States of America,2009,106:17588-17593.
- [16]VLAD F,RUBIO S,RODRIGUES A,SIRICHANDRA C,BE-LIN C,ROBERT N,LEUNG J,RODRIGUEZ P L,LAURIEREC,MERLOT S.Protein phosphatases 2C regulate the activationof the Snf1-related kinase OST1 by abscisic acid in Arabidopsis[J].The Plant Cell,2009,21:3170-3184.
- [17]YIN P,FAN H,HAO Q,YUAN X,WU D,PANG Y,YAN C,LI W,WANG J,YAN N.Structural insights into the mechanismof abscisic acid signaling by PYL proteins[J].Nature Structuraland Molecular Biology,2009,16:1230-1236.
- [18]SZOSTKIEWICZ I,RICHTER K,KEPKA M,DEMMEL S,MAY,KORTE A,ASSAAD F F,CHRISTMANN A,GRILL E.Close-ly related receptor complexes differ in their ABA selectivity andsensitivity[J].The Plant Journal,2010,61:25-35.
- [19]SAAVEDRA X,MODREGO A,RODRIGUEZ D,GONZALEZ-GARCIA M P,SANZ L,NICOLAS G,LORENZO O.The nucle-ar interactor PYL8/RCAR3 of Fagus sylvatica Fs PP2C1 is a posi-tive regulator of abscisic acid signaling in seeds and stress[J].Plant Physiology,2010,152:133-150.
- [20]MA Y,SZOSTKIEWICZ I,KORTE A,MOES D,YANG Y,CHRISTMANN A,GRILL E.Regulators of PP2C phosphatase activity function as abscisic acid sensors[J].Science(New York,NY),2009,324:1064-1068.
- [21]OKUBO M,FURUKAWA Y,SAKURATANI T.Growth,flower-ing and leaf properties of pear cultivars grafted on two Asian pearrootstock seedlings under Na Cl irrigation[J].Scientia Horticul-turae,2000,85:91-101.
- [22]NAMBARA E,MARION P A.Abscisia acid biosynthesis andca-tabolism[J].Annu Rev Plant Biol,2005,56:165-185.
- [23]BURBIDGE A,GRIEVE T M,JACKSON A.Characterizationofthe ABA-deficient tomato mutant notabilis andits relationshipwith maize Vp14[J].Plant J,1999,17:427-431.
- [24]LU Ping,TIAN Yue-sheng,WANG Ming-xue,LI Shan,ZHAOJing-ya.Cloing and characterization of 9-cis-eposycarotenoid di-oxygenase gene(NCED)encoding a key enzyme during abscisicacid biosynthesis in Lycium barbarum L[J].Journal of Plant Genet-ic Resources,2013,14(2):303-310.陆平,田跃胜,王名雪,李杉,赵静雅.枸杞脱落酸生物合成关键酶基因NCED的克隆及表达分析[J].植物遗传资源学报,2013,14(2):303-310.
- [25]HAN Jin-long,LI Hui,CONG Yu,CHANG You-hong,LINJing.Comparison of tow CBL genes on stress tolerance functionsfrom Pyrus betulaefolia Bunge[J].Journal of Fruit Science,2014,31(4):529-535.韩金龙,李慧,丛郁,常有宏,蔺经.杜梨CBL1和CBL7基因对非生物逆境的响应[J].果树学报,2014,31(4):529-535.
- [26]LIU X,YUE Y,LI B,NIE Y,LI W,WU W H,MA L.A G pro-tein-coupled receptor is a plasma membrane receptor for theplant hormone abscisic acid[J].Science(New York,NY),2007,315:1712-1716.
- [27]SHEN Y Y,WANG X F,WU F Q,DU S Y,CAO Z,SHANGY,WANG X L,PENG C C,YU X C,ZHU S Y,FAN R C,XUY H,ZHANG D P.The Mg-chelatase H subunit is an abscisic ac-id receptor[J].Nature,2006,443:823-826.
- [28]ADIE B A,PEREZ-PEREZ J,PEREZ-PEREZ M M,GODOYM,SANCHEZ-SERRANO J J,SCHMELZ E A,SOLANO R.ABA is an essential signal for plant resistance to pathogens affect-ing JA biosynthesis and the activation of defenses in Arabidopsis[J].The Plant cell,2007,19:1665-1681.
- [29]DING Z,LI S,AN X,LIU X,QIN H,WANG D.Transgenic ex-pression of MYB15 confers enhanced sensitivity to abscisic acidand improved drought tolerance in Arabidopsis thaliana[J].Jour-nal of Genetics and Genomics,2009,36:17-29.
- [30]SANTIAGO J,RODRIGUES A,SAEZ A,RUBIO S,ANTONIR,DUPEUX F,PARK S Y,MA'RQUEZ J A,CUTLER S R,RODRIGUEZ P L.Modulation of drought resistance by the abscis-ic acid-receptor PYL5 through inhibition of clade A PP2Cs[J].The Plant Journal,2009,60:575-588.
- [31]IUCHI S,KOBAYASHI M,TAJI T,NARAMOTO M,SEKI M,KATO T,TABATA S,KAKUBARI Y,YAMAGUCHI-SHINO-ZAKI K,SHINOZAKI K.Regulation of drought tolerance bygene manipulation of 9-cisepoxycarotenoid dioxygenase,a keyenzyme in abscisic acid biosynthesis in Arabidopsis[J].The PlantJournal,2001,27:325-333.
- [32]LI Kang,NIE Xiao-jun,FANG Gui-ying,WANG Le,YUEHua,SHE Kui-jun,SONG Wei-ning.Cloning and expression ofa NCED gene in wheat and its relative species[J].Acta Agricul-turae Boreali-occidental is Sinica,2010,19(6):55-59.李康,聂小军,方桂英,王乐,岳华,佘奎军,宋卫宁.普通小麦及其近缘种NCED基因的克隆及其表达分析[J].西北植物学报,2010,19(6):55-59.