茉莉酸刺激的橡胶树胶乳cDNA消减文库的构建及其序列分析  

Construction of cDNA Library and Analyses of cDNA Sequences in Latex from JA-Stimulated Rubber Trees

作　　者：曾日中[1] 段翠芳[1] 黎瑜[1] 郝秉中[1] 

机构地区：[1]中国热带农业科学院橡胶研究所,农业部热带作物栽培生理学重点实验室,海南儋州571737

出　　处：《热带作物学报》2003年第3期1-6,共6页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金资助项目(编号30260088)

摘　　要：采用抑制性消减杂交(suppression subtractive hybridization,SSH)技术,构建了外源茉莉酸刺激条件下橡胶树胶乳与未处理橡胶树胶乳差异表达的cDNA消减文库,经蓝白斑筛选共得到121个含有插入片段的阳性克隆。通过菌落PCR的方法分别对阳性克隆的插入片段进行扩增,结果表明,95％左右阳性克隆插入片段的大小在200-600 bp之间。随机选取25个克隆进行测序,并对所得的25条表达序列标签(EST)序列用Blastn(基本局域联配搜寻工具)检索基因文库(genbank),其中10条EST片断可找到碱基序列相似性大于80％以上的同源基因序列,其余15条EST为没有任何功能线索的未知序列。外源茉莉酸刺激条件下橡胶树胶乳cDNA消减文库的构建和在此基础上克隆橡胶树胶乳中JA信号的候选应激基因,将为开展茉莉酸调控橡胶树胶乳代谢和橡胶生物合成的分子机理研究打下基础。The SSH library of differently expressed cDNA was constructed in the latex from the JA-stimulated rubber tree to study the latex metabolism and molecular mechanism of rubber biosynthesis regulated by exogenous jasmonic acid (JA) in the rubber tree (Hevea brasiliensis). A total of 121 positive clones were obtained by screening the blue-white conoly. The inserted cDNAs of these positive clones were amplified by colony PCR. The results showed that the inserted cDNAs of 95 % positive clones were from 200 bp to 600 bp. After sequencing of 25 random clones picked up from the SSH library and searching Genbank with Blastn, ten ESTs of the 25 random clones had homology (>80 %) with cDNA sequences of the Genbank, while the other 15 ESTs were unknown sequences. They may be the peculiar functional genes of rubber tree.

关 键 词：橡胶树 胶乳 抑制性消减杂交 CDNA消减文库 茉莉酸 乳管细胞 生物合成 植物激素 信号转导 

分 类 号：S794.1[农业科学—林木遗传育种]