杜梨2个MYB转录因子基因的克隆及序列分析Cloning and sequence analysis of two MYB transcription factors in Pyrus betulifolia
冉昆;王少敏;董放;魏树伟;张勇;王宏伟;
摘要(Abstract):
【目的】克隆杜梨(Pyrus betulifolia Bunge)MYB家族的2个基因(Pb3RMYB和Pb4RMYB),分析其序列特征,为研究其调控的分子机制奠定基础。【方法】利用RT-PCR和RACE技术克隆杜梨Pb3RMYB和Pb4RMYB的c DNA序列全长;利用BLAST和ORF Finder对核酸序列进行分析,利用CDD、Prot Param、TMHMM、Signal P、Plant-m Ploc、SPOMA、DANMAN、MEGA6等软件对氨基酸序列进行分析。【结果】克隆得到Pb3RMYB和Pb4RMYB基因全长,Gen Bank登录号分别为KX272614和KX272615。2者均含有MYB保守结构域,二级结构均以无规则卷曲、α-螺旋、延伸链和β-折叠为主,亚细胞定位分析表明2者均定位于细胞核中。其中,Pb3RMYB属于3R-MYB亚家族,全长2 355 bp,ORF长1 710bp,编码569个氨基酸,蛋白分子质量约为62 077.3 Da,等电点为7.08;BLAST分析表明Pb3RMYB与已报道的其他物种3RMYB具有较高的相似度。Pb4RMYB属于4R-MYB亚家族,全长3 625 bp,ORF长2 913 bp,编码970个氨基酸,蛋白分子质量约为109 600.6 Da,等电点为7.41。【结论】获得了2个杜梨MYB基因Pb3RMYB和Pb4RMYB,分别属于3RMYB和4R-MYB亚家族。
关键词(KeyWords): 杜梨;MYB转录因子;基因克隆;序列分析
基金项目(Foundation): 山东省自然科学基金(ZR2015YL075);; 山东省农业科学院青年科研基金(2015YQN41);; 山东省良种工程项目(梨优质矮化、多抗功能基因挖掘与种质创新利用2014);; 现代农业产业技术体系建设专项(CARS-29-31)
作者(Author): 冉昆;王少敏;董放;魏树伟;张勇;王宏伟;
Email:
DOI: 10.13925/j.cnki.gsxb.2016.S.06
参考文献(References):
- [1]ALLAN A C,HELLENS R P,LAING W A.MYB transcription factors that colour our fruit[J].Trends in Plant Science,2008,13(3):99-102.
- [2]DUBOS C,STRACKE R,GROTEWOLD E,WEISSHAAR B,MARTIN C,LEPINIEC L.MYB transcription factors in Arabidopsis[J].Trends in Plant Science,2010,15(10):573-581.
- [3]FELLER A,MACHEMER K,BRAUN E L,GROTEWOLD E.Evolutionary and comparative analysis of MYB and b HLH plant transcription factors[J].The Plant Journal,2011,66(1):94-116.
- [4]王春荣,王庆杰,李晓玲,姚玉新,郝玉金.葡萄中盐诱导的R2R3-MYB基因的筛选与表达分析[J].园艺学报,2014,41(3):529-535.WANG Chunrong,WANG Qingjie,LI Xiaoling,YAO Yuxin,HAO Yujin.Screening and expression analysis of salt-induced R2R3-MYB genes in grapevine[J].Acta Horticulturae Sinica,2014,41(3):529-535.
- [5]AMBAWAT S,SHARMA P,YADAV N R,YADAV R C.MYB transcription factor genes as regulators for plant responses:an overview[J].Physiology and Molecular Biology of Plants,2013,9(3):307-321.
- [6]BALDONI E,GENGA A,COMINELLI E.Plant MYB transcription factors:their role in drought response mechanisms[J].International Journal of Molecular Sciences,2015,16(7):15811-15851.
- [7]LI C,NG C K Y,FAN L M.MYB transcription factors,active players in abiotic stress signaling[J].Environmental and Experimental Botany,2015,114:80-91.
- [8]CHEN Y H,YANG X Y,HE K,LIU M H,LIU J,GAO Z F,LIN Z Q,ZHAO Y F,WANG X X,QIU X M,SHEN Y P,ZHANG L,DENG X H,LUO J C,DENG X W,CHEN Z L,GU H Y,QU L J.The MYB transcription factor superfamily of Arabidopsis:Expression analysis and phylogenetic comparison with the rice MYB family[J].Plant Molecular Biology,2006,60(1):107-124.
- [9]JUNG C,SEO J S,HAN S W,KOO Y J,KIM C H,SONG S I,NAHM B H,DO CHOI Y,CHEONG J J.Overexpression of AtMYB44 enhances stomatal closure to confer abiotic stress tolerance in transgenic Arabidopsis[J].Plant Physiology,2008,146(2):623-635.
- [10]AGARWAL M,HAO Y,KAPOOR A,DONG C H,FUJII H,ZHENG X,ZHU J K.A R2R3 type MYB transcription factor is involved in the cold regulation of CBF genes and in acquired freezing tolerance[J].The Journal of Biological Chemistry,2006,281(49):37636-37645.
- [11]DAI X,XU Y,MA Q,XU W,WANG T,XUE Y,CHONG K.Overexpression of an R1R2R3 MYB gene,Os MYB3R-2,increases tolerance to freezing,drought,and salt stress in transgenic Arabidopsis[J].Plant Physiology,2007,143(4):1739-1751.
- [12]CAO Y,HAN Y,LI D,LIN Y,CAI Y.MYB transcription factors in Chinese pear(Pyrus bretschneideri Rehd.):genome-wide identification,classification,and expression profiling during fruit development[J].Front in Plant Science,2016,7:577.
- [13]宗宇,孙萍,牛庆丰,滕元文.中国北方野生杜梨分布现状及其形态多样性评价[J].果树学报,2013,30(6):918-923.ZONG Yu,SUN Ping,NIU Qingfeng,TENG Yuanwen.Distribution situation and assessment of morphological diversity of wild Pyrus betulaefolia in Northern China[J].Journal of Fruit Science,2013,30(6):918-923.
- [14]RAN K,YANG H Q,SUN X L,LI Q,JIANG Q Q,ZHANG W W,SHEN W.Isolation,characterization,and structure analysis of a vacuolar processing enzyme gene(Mh VPEγ)from Malus hupehensis(Pamp)Rehd[J].Applied Biochemistry and Biotechnology,2014,173(2):579-595.
- [15]LETUNIC I,DOERKS T,BORK P.SMART:recent updates,new developments and status in 2015[J].Nucleic Acids Research,2015,43(D1):D257-D260.
- [16]MARCHLER-BAUER A,ZHENG C,CHITSAZ F,DERBYSHIRE M K,GEER L Y,GEER R C,GONZALES N R,GWADZ M,HURWITZ D I,LANCZYCKI C J,LU F,LU S,MARCHLER G H,SONG J S,THANKI N,YAMASHITA R A,ZHANG D,BRYANT S H.CDD:conserved domains and protein three-dimensional structure[J].Nucleic Acids Research,2013,41(D1):348-352.
- [17]WILKINS O,NAHAL H,FOONG J,PROVART N J,CAMPBELL M M.Expansion and diversification of the Populus R2R3-MYB family of transcription factors[J].Plant Physiology,2009,149(2):981-993.
- [18]左然,徐美玲,柴国华,周功克.植物MYB转录因子功能及调控机制研究进展[J].生命科学,2012,24(10):1133-1140.ZUO Ran,XU Meiling,CHAI Guohua,ZHOU Gongke.Function and regulation mechanism of plant MYB transcription factors[J].Chinese Bulletin of Life Sciences,2012,24(10):1133-1140.
- [19]刘蕾,杜海,唐晓凤,吴燕民,黄玉碧,唐益雄.MYB转录因子在植物抗逆胁迫中的作用及其分子机理[J].遗传,2008,30(10):1265-1271.LIU Lei,DU Hai,TANG Xiaofeng,WU Yanmin,HUANG Yubi,TANG Yixiong.The roles of MYB transcription factors on plant defense responses and its molecular mechanism[J].Hereditas,2008,30(10):1265-1271.