月季NBS-LRR-RGAs的分离与鉴定  

Isolation and characterization of NBS-LRR resistance gene candidates in rose

作　　者：张非亚 杜运鹏[2] 刘瑞峰[1] 袁晓娜[1] 张冬梅 贾桂霞[1] 

机构地区：[1]花卉种质创新与分子育种北京市重点实验室,国家花卉工程技术研究中心,城乡生态环境北京实验室,北京林业大学园林学院 [2]北京农业生物技术研究中心,北京市农林科学院 [3]上海市园林科学规划研究院

出　　处：《北京林业大学学报》2016年第11期89-96,共8页Journal of Beijing Forestry University

基　　金：“863”国家高技术研究发展计划项目(2011AA100208);上海科委农业重点项目(13391900902)

摘　　要：RGAs已经在很多植物中得到分离与克隆,它往往是R基因或者是R基因的一部分。为了分离与鉴定与月季抗黑斑病相关的RGAs,在转录组数据基础上,设计简并引物6对,从现代月季、疏花蔷薇、‘白玫瑰’上分离克隆了NBS-LRR-RGAs,挑取750个克隆进行序列分析。从系统发育树可以看出:所克隆的RGAs与已克隆的其他植物的R基因具有较近的亲缘关系,分为non-TIR-NBS-RGAs和TIR-NBS-RGAs 2个大类,10个进化枝,其中所克隆得到的non-TIR-NBS-RGAs的比例远大于TIR-NBS-RGAs的比例;氨基酸序列比对发现所克隆的RGAs均含有完整的NBS结构域P-Loop、kinase-2、kinase-3a和GLPL;通过进化树对不同家族的6个RGAs的表达分析可看出,它们都不同程度、不同时间地参与了抗病反应。RGAs have been isolated and characterized from many different plant species,and they are often located in genomic regions containing R genes. For the isolation and identification of RGAs,which were related to rose resistance to black pot,six pairs of degenerate primers were designed based on the transcriptome sequencing,and the NBS-LRR-RGAs were isolated and cloned from modern rose,Rosa laxa and Rosa rugosa ‘White＇. 750 clones were selected and sequenced. Phylogenetic studies revealed that the sequenced rose RGAs clones showed a close relationship with R genes and these sequences were classified into ten clades,with non-TIR-NBS-RGAs and TIR-NBS-RGAs subclasses. The proportion of cloned non-TIR-NBS-RGAs was far greater than that of TIR-NBS-RGAs. The deduced amino acid sequences of RGAs all contained the whole conserved domains： P-Loop,kinase-2,kinase-3a and GLPL.The analysis of quantitative real-time PCR shows that the six RGAs from different clade participate in the reaction in different transcript levels and different time.

关 键 词：月季 黑斑病 NBS-LRR-RGAs 

分 类 号：S685.12[农业科学—观赏园艺]