不结球白菜S位点受体激酶基因片段的克隆与表达分析  

Cloning and expression analysis of SRK gene fragment in non-heading Chinese cabbage

作　　者：张爱芬[1] 王立[1] 侯喜林[1] 刘同坤[1] 李英[1] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室/农业部南方蔬菜遗传改良重点开放实验室,江苏南京210095

出　　处：《南京农业大学学报》2011年第3期25-30,共6页Journal of Nanjing Agricultural University

基　　金：国家973计划项目子课题(2009CB119001-04);教育部新世纪优秀人才支持计划项目(NCET-07-0439)

摘　　要：以不结球白菜自交不亲和系03的基因组DNA和柱头cDNA为模板,利用引物SRKF/SRKR扩增获得长度为964 bp和646 bp的SRK基因片段。序列比较分析表明,克隆的基因片段属于SRK基因的激酶域,该序列包含4个外显子和3个内含子,编码215个氨基酸,与芜菁、甘蓝、芥蓝等SRK基因有90%以上的相似性。荧光定量PCR分析结果表明:自交不亲和系03和自交亲和系105不同组织中SRK基因的表达水平存在显著的差异,SRK基因主要在自交不亲和系的柱头中高度表达,自交亲和系中该基因的表达主要分布于叶片、花蕾和柱头组织中。Using primers SRKF/SRKR,964 bp gDNA（BcSRK-1）and 646 bp cDNA sequences of S locus receptor kinase gene were obtained from self-incompatible line 03 of Brassica campestris ssp.chinensis Makino.Sequence alignment and phylogenetic analysis revealed that DNA and cDNA of BcSRK-1 belonged to kinase domain,containing 4 exons and 3 introns,and coded 215 amino acids.Furthermore,its similarity was over 90% with the SRK gene of Brassica rapa,Brassica oleracea and Brassica oleracea var.alboglabra.Real-time PCR analysis showed there was a significant difference between self-incompatible and self-compatible lines.BcSRK-1 was highly expressed in stigmas of self-incompatible line.Howerer,in self-compatible line,BcSRK-1 was expressed in an even lower level whether in stigmas,buds or leaves.

关 键 词：不结球白菜 S位点受体激酶基因 荧光定量PCR 基因表达分析 

分 类 号：S634.3[农业科学—蔬菜学]