青稞抗黄矮病抑制差减文库的构建及差异表达基因的分离  

Construction of a Naked Barley SSH cDNA Library Against BYDV and Isolation of Different Expression Genes

作　　者：梅红[1] 赖建华[2] 何婕[1] 木德伟 施晓群[1] 

机构地区：[1]云南省农业科学院农业环境资源研究所,云南昆明650205 [2]云南大学生命科学学院,云南昆明650031 [3]迪庆州农业科学研究所,云南香格里拉674400

出　　处：《西南农业学报》2009年第2期319-323,共5页Southwest China Journal of Agricultural Sciences

基　　金：云南省科技厅省青年基础研究项目(2004C0026Q)

摘　　要：本研究应用抑制差减杂交方法构建了感染黄矮病毒早期,高抗病和高感病青稞种质的差减cDNA文库,共获得了143个带有插入片断的克隆。以构建的抗黄矮病抑制差减文库的62个片段为基础,对抗病青稞品种52号和感病青稞品种品引5号在感染黄矮病后24 h的差异表达基因进行了杂交筛选,获得了9个差异表达的基因片段。对这些片段进行序列分析和同源对比,获得了5个抗性相关基因片段。片段U3,U24在感染后24 h的抗性品系中表达增强,同时在EST序列比对中没有发现同源序列,可能为新的抗性相关基因。片段U58,U62为分别在抗性和感性品种中高表达的叶绿体相关基因。片段U23为多胺代谢通路相关基因。通过对这些基因片段的分析,对青稞感染黄矮病早期抗性的机制进行了初步的探索。A suppression subtractive hybridization （SSH） cDNA library was constructed with naked barley leaf induced by Barley yellow dwarf virus, and the process of library construction was checked and analyzed. The results showed that the technique was efficient in the subtractive hybridization. 143 clones with insert fragments were obstained. 62 fragments in this suppression subtractive hybridization （SSH） cDNA library was hybridized with cDNA of one high resistance cultivars and one high sensitive cultivars infected by BYDV. 9 fragment showed different expression in sensitive and resistance cultivars. 2 fragments expressed in resistance cultivars specially were new gene fragment with- out the similar EST sequence in Genbank. 2 fragments expressed in sensitive and resistance cultivars were related to chloroplast, respectively. 1 fragment expressed in sensitive cultivars was S-adenosylmethionine decarboxylase resulting in polyamine disruption and growth perturbations. These analyses provided some clues for the anti-BYDV mechanism of naked barley.

关 键 词：青稞 黄矮病 差减杂交文库 

分 类 号：S512.3[农业科学—作物学]