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

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.