黑皮冬瓜NBS类抗病基因同源序列克隆与分析  

Cloning and Sequence Analysis of NBS Type Resistant Gene Analogs from Black Wax Gourd(Benincasa hispida)

作　　者：赵芹[1] 谢大森[1] 何晓明[1] 罗少波[1] 李明珠[1] 

机构地区：[1]广东省农业科学院蔬菜研究所,广东广州510640

出　　处：《热带作物学报》2014年第10期2071-2077,共7页Chinese Journal of Tropical Crops

基　　金：国家自然科学青年基金(No.31311643);广东省自然科学基金(No.10451064001006063);广州市珠江科技新星项目(No.2013086)

摘　　要：以黑皮冬瓜＂B98K＂种质为试材,根据已克隆的植物NBS类抗病基因保守结构域设计简并引物,PCR扩增冬瓜基因组DNA,获得对应区段的DNA片段,回收克隆与测序后,得到19条抗病同源序列（命名为DNB1至DNB19）。利用DNAStar软件进行序列分析及同源性比对发现,这些抗病同源序列长度在249-252 nt之间,其核苷酸序列同源性在48.4%-98.8%之间,氨基酸序列同源性为23.2%-100.0%,推导氨基酸序列具有P-loop（GGVGKTT）、Kinase-2a（VLDDVW）典型的NBS保守结构域。同源进化分析将其全部归为non TIR-NBS-LRR类,与推导氨基酸序列多重比较结果一致;该序列与其他作物已克隆抗病基因的氨基酸序列同源性在24.7%-99.0%之间。该组序列已登录Genbank,登录号为KF776401-KF776419。According to the conserved domains of NBS typ e disease-resistant genes in most known plants,two degenerate primers were designed and synthesized.Nineteen RGAs named from DNB1 to DNA19 were obtained by PCR amplification of genome DNA from black wax gourd ＂ B98K＂ variety.Sequences analysis and alignment results indicated that full-length of these RGAs varied from 249 nt to 252 nt,the deduced amino acids sequences contained typical conserved domains of NBS family,such as P-loop（GGVGKTT） and Kinase-2a（VLDDVW）.The Gen Bank accession numbers were obtained（from KF776401 to KF776419）.These RGAs showed great homologous differences with the similarity from 48.4% to 98.8% and with genetic divergence from 1.2 to 88.9 in nucleotide sequence level,while the amino acid sequence homology varied from 23.2% to 100.0%.These RGAs were all divided into non TIR-NBS-LRR group,which was consistent with the multiple-alignment of deduced amino acid sequences.The similarities varied from 24.7% to 99.0% compared with NBS RGAs of other known species.

关 键 词：冬瓜 NBS 抗病同源序列 同源克隆 序列分析 

分 类 号：S642[农业科学—蔬菜学]