萝卜过氧化物酶基因片段的cDNA克隆及花期表达谱分析  

cDNA Partial Cloning of Peroxidases Genes from Raphanus sativus L.and Their Expression in Floral Stage

作　　者：王林嵩[1] 李小娟[1] 李芳军[1] 孙向东[1] 王丽[1] 宗秀征[1] 杨少华[1] 王一帆[1] 

机构地区：[1]河南师范大学生命科学学院,新乡453007

出　　处：《中国蔬菜》2009年第6期14-20,共7页China Vegetables

基　　金：国家自然科学基金(30570135)

摘　　要：通过研究过氧化物酶(POD)活性在心里美萝卜花期的变化以及萝卜POD基因的结构,探讨POD的基因结构和功能之间的关系。测定了萝卜花期不同时间各个部位中POD活性,用改进的半定量RT-PCR分析了过氧化物酶基因(Rsprx)的表达谱,并通过RT-PCR克隆过氧化物酶基因的cDNA片段。结果表明,心里美萝卜花期不同器官中的POD活性具有明显的变化,地上部分和地下部分POD活性差异显著,在结荚期的木质部Ⅱ中达到峰值(17.53±0.35)×103U·g-1(FW)。不同器官中Rsprx的表达差异明显,花蕾和荚中表达最强。从花蕾和荚中得到4个长度为400bp左右的cDNA克隆片段,序列比对结果显示,与棉花的花器官过氧化物酶基因Ghpod具有较高的同源性和结构相似性。因此认为POD与萝卜花期生长发育有一定的关系,且有多种过氧化物酶基因在该过程中表达。In order to explore the relationship between the structure and function of peroxidases, the changes of radish peroxidases （POD） activities were investigated at floral stage and the POD gene structure were also analyzed. Activities of POD in different organs of radish were determined. The expression profile of radish peroxidase genes （Rsprx） were analyzed by modified semi-quantitive RT - PCR, some partial cDNAs of radish peroxidase genes were cloned and sequenced. The results showed that the activities of POD were changed greatly in difterent organs at florescence in the Chinese red radish. POD activities were significantly difference between above-ground and under-ground organs. The peak of the POD activity was （17.53±0.35）×10^3U·g^-1 （FW） in xylem Ⅱ at producing legumen stage. The expression levels of the Rsprx were higher among different organs of Chinese red radish. There were significant differences in expression of Rsprx in bud and legumen. Four radish pcroxidase partial cDNAs about 400 bp were cloned from the bud and legumen, which showed high identity and structure similarity with the flower-specific expressed cotton gene Ghpod by sequence alignment. There are correlation between POD and florescence growth and development of radish, and various peroxidases genes expression during the floral organ develop-ment processes of radish.

关 键 词：萝卜 过氧化物酶 RT-PCR CDNA克隆 

分 类 号：S631.1[农业科学—蔬菜学]