黑斑病菌诱导的紫薯抑制性差减杂交文库构建及ESTs分析  

Construction of SSH cDNA Library of Sweet Potato Induced by Black Rot and It's ESTs Analysis

作　　者：曾绍华[1] 张聪[2] 李明[2] 屈会娟[2] 张敏[1] 阎文昭[2] 蒲志刚[2] 

机构地区：[1]四川农业大学,四川成都611130 [2]四川省农业科学院生物技术核技术研究所,四川成都610066

出　　处：《西南农业学报》2014年第1期53-58,共6页Southwest China Journal of Agricultural Sciences

基　　金：成都市科技计划项目(11DXYB238NC-027)

摘　　要：本研究以紫薯1号为材料,运用抑制性差减杂交(SSH)技术,构建了经黑斑病菌诱导12、24、36、48、60 h后的甘薯块根混合SSH cDNA文库。随机挑取103个cDNA克隆进行5'端测序,共获得87个非重复序列,其中包括29个重叠群,58个独立的ESTs,利用tblastx对87个序列在NCBI数据库中进行同源性比对。结果表明,在编号为14的EST中找到了"蔷薇"黑斑抗性muRdr1基因位点,基因全序列包括muRdr1A、muRdr1B、muRdr1C、muRdr1D、muRdr1E、muRdr1F、muRdr1G、muRdr1H、muRdr1I共9个基因。另外有2类基因的含量较高,一类是紫薯组织特异性相关基因,另一类是紫薯受干旱胁迫的抗逆相关基因。构建cDNA文库,应用EST技术并结合生物信息学技术,对紫薯抗病相关基因的表达分析,从基因水平上来研究紫薯抗病机制,为今后的紫薯抗病遗传育种及抗病机理奠定论基础。In this paper,taken purple sweet potato No.1 as tested materials,the mixed SSH library was built after the purple sweet potato tuberous roots were induced by black rot in 12,24,36,48,60 h.103 clones were randomly picked out and sequenced,and a total of 87 ESTs,including 29 contigs and 58 singlets,were obtained.These 87 ESTs were analyzed with tblastx compared with NCBI database sequences homologous.The result showed that the spot of Rose black resistance （muRdr1 gene locus and gene sequences） were found in No.14 EST,including a total of 9 genes （muRdr1 A,muRdr1 B,muRdr1 C,muRdr1 D,muRdd E,muRdr 1 F,muRdr1 G,muRdr 1 H,muRdrl I）.In addition,there were higher levels fo two types of genes,one was the related genes of purple sweet potato tissue specificity,and the other was the related genes of purple sweet potato resistance to drought stress.The suppression subtractive hybridization（SSH） technology built cDNA library,using EST technology combined with bioinformatics technology,the expreessions of related disease resitance genes from the genetic level were analyzed to provide theoretical basis for the black rot resistant mechanism of purple sweet potato.

关 键 词：紫薯 黑斑病菌诱导 抑制差减杂交技术 表达序列标签 

分 类 号：S435.311[农业科学—农业昆虫与害虫防治]