萝卜RsSRK-a基因cDNA克隆与表达特性分析  

The full length cDNA cloning of RsSRK-a gene and its expression analysis in radish (Raphanus sativus L.)

作　　者：周艳孔[1] 邵文婷[1] 龚义勤[1] 朱献文[2] 马二磊[1] 赵天荣[1] 柳李旺[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/园艺学院,江苏南京210095 [2]北达科达州立大学植物科学系

出　　处：《南京农业大学学报》2010年第4期13-18,共6页Journal of Nanjing Agricultural University

基　　金：教育部新世纪优秀人才支持计划项目(NCET-10-0473);国家863计划项目(2008AA10Z150)

摘　　要：以萝卜Nau-DY06为试材,采用亲和指数和荧光显微镜进行自交不亲和性测定,结果表明其为稳定自交不亲和系。根据不同作物SRK基因保守序列设计特异引物,以Nau-DY06柱头为材料,采用RT-PCR扩增获得SRK基因cDNA序列,包含2562bp开放阅读框(ORF),命名为RsSRK-a(GenBank登录号:GQ121139)。该基因cDNA序列编码853个氨基酸,其核苷酸序列与甘蓝型油菜SRK3、羽衣甘蓝SRK13-b基因同源性均为89%。RsSRK-a基因序列编码的蛋白质相对分子质量为96.6×103,等电点为6.69。半定量RT-PCR分析表明,RsSRK-a基因在自交不亲和系花期柱头中表达量远远高于蕾期柱头,而在植株的其他部位均不表达,表明SRK基因与自交不亲和特性表达密切相关。The radish self-incompatibility line Nau-DY06 was identified with the compatibility index and the fluorescence microscopy method.An ORF encoding 853 amino acids was amplified by RT-PCR from cDNA of the stigma of Nau-DY06 with the primer derived from the conserved sequences of the SRK genes in different crops.The gene was designated as RsSRK-a （GenBank accession number：GQ121139）.Its identity with the SRK3 in Brassica rapa,the SRK13-b in B.oleracea var.acephala,was 89% respectively.The molecular weight of the predicated protein encoded was 96.6 × 103 and its theoretical pI was 6.69.Semi-quantitative RT-PCR analysis showed that expression of the RsSRK-a was detectable in the stigma of two days before anthesis and especially high at anthesis stage.Expression profiling of the RsSRK-a in radish stigma indicated that it might play an important role in the selfincompatibility.The results provide important information for the exploration of the mechanism and utilization of self-incompatibility in radish.

关 键 词：萝卜 自交不亲和性 SRK基因 克隆 表达分析 

分 类 号：S631.1[农业科学—蔬菜学]