青花菜CC-NBS-LRR抗病基因BoCNL1的克隆与分析  

Cloning and characterization of a CC-NBS-LRR disease resistance gene of BoCNL1 from Brassica oleracea var. italica

作　　者：黄笑梅[1] 金建峰[2] 张雪[1] 朱思眉 朱柯柯 赵罗鹏 蒋明[1] 

机构地区：[1]台州学院生命科学学院,浙江椒江318000 [2]浙江大学生命科学学院,浙江杭州310058

出　　处：《浙江农业学报》2016年第2期259-263,共5页Acta Agriculturae Zhejiangensis

基　　金：浙江省自然科学基金项目(LY13C150003);浙江省大学生科技创新活动计划(新苗人才计划)(2014R428011);浙江省本科院校中青年学科带头人学术攀登项目(pd2013420)

摘　　要：根据已知序列设计PCR引物,从青花菜中克隆1个NBS-LRR(核苷酸结合位点-富含亮氨酸重复)抗病基因BoCNL1;在生物信息学分析的基础上,利用RT-PCR研究该基因在不同器官中的表达模式。测序结果表明,BoCNL1基因的编码区全长为2 550 bp,编码849个氨基酸;编码蛋白具CC(卷曲螺旋)、NBS和LRR结构域;进化分析结果表明,BoCNL1与不结球白菜的关系最近,在进化树上处于同一分支,与醉蝶花的关系最远;RT-PCR结果表明,BoCNL1在根、花茎、叶、花蕾、开放的花和嫩角果中均有表达,但表达量低。Primer pairs were designed according to known sequences,and a nucleotide-binding site plus leucine-rich repeat（ NBS-LRR） disease resistance gene,designated BoCNL1,was isolated from broccoli（ Brassica oleracea var.italic）. Bioinformatic analysis were performed,and RT-PCR was used to reveal expression patterns of BoCNL1 in different organs. Results indicated that the complete coding sequence of BoCNL1 was 2 550 bp in length,encoding849 amino acids; and the deduced protein sequence contained coiled coil（ CC）,NBS,and LRR domains. Phylogenetic analysis results showed BoCNL1 was grouped with the homologous gene in B. rapa,indicating their closest relationship,and the longest genetic distance was observed between B. oleracea var. italica and Tarenaya hassleriana.RT-PCR results demonstrated that BoCNL1 expressed with low levels of transcripts in roots,flower stalks,leaves,flower buds,flowers,as well as young siliques.

关 键 词：青花菜 抗病基因 CC-NBS-LRR 基因克隆 

分 类 号：S635.3[农业科学—蔬菜学] Q78[农业科学—园艺学]