橡胶树HbGAIP-B基因的克隆与表达分析  

Molecular Cloning and Expression of HbGAIP-B Gene in Rubber Tree(Hevea brasiliensis Müll. Arg.)

作　　者：吴绍华[1] 郝泽芸 张世鑫[1] 邓小敏[1] 田维敏[1] 

机构地区：[1]中国热带农业科学院橡胶研究所,农业部橡胶树生物学与遗传资源利用重点实验室,省部共建国家重点实验室培育基地-海南省热带作物栽培生理学重点实验室,海南儋州571737 [2]海南大学应用科技学院,海南儋州571737

出　　处：《热带作物学报》2016年第5期881-887,共7页Chinese Journal of Tropical Crops

基　　金：国家天然橡胶产业技术体系(No.CARS-34-GW1);国家自然科学基金(No.31300504)

摘　　要：DELLA是赤霉素信号传导途径中一类重要的阻遏蛋白。本研究通过RT-PCR技术,从橡胶树优良品种‘热研7-33-97’胶乳中克隆了1个DELLA蛋白编码基因(GenBank登录号为KT696440)。该基因全长cDNA序列2135bp,阅读框1905bp,编码634个氨基酸,其编码蛋白含有DELLA和GRAS结构域,分子量为69.89ku,理论等电点为5.50。HbGAIP-B氨基酸序列与麻疯树JcGAIP-B、蓖麻RcGAIP-B、拟南芥AtRGA、AtGAI、AtRGL1、AtRGL2和AtRGL3的相似性分别为82.43%、78.18%、65.05%、61.73%、52.28%、53.51%和49.92%。进化树分析表明,HbGAIP-B与JcGAIP-B亲缘关系最近。定量PCR分析表明,HbGAIP-B的表达受割胶处理诱导下调。赤霉素和乙烯利处理4h显著上调HbGAIP-B的表达,茉莉酸甲酯处理4h小幅下调HbGAIP-B表达。DELLA are kind of repressor proteins of GA signaling. In this work, a full length cDNA sequence of HbGAIP-B （GenBank accession no. KT696440） was obtained from the latex of rubber tree clone ＇CATAS7-33-97＇ by RT-PCR. HbGAIP-B was 2 135 base pairs （bp） in length and contained an open reading frame （ORF） of 1 642 bp. The ORF encoded 613 amino acid residues, which contained DELLA and GRAS domain, with a predicted molecular mass of 66.476 ku and a pI of 5.19. The deduced amino acid sequences of HbGAIP-B showed high identities of 82.43%, 78.18%, 65.05%, 61.73%, 52.28%, 53.51% and 49.92% to JcGAIP-B, RcGAIP-B, AtRGA, AtGAI, AtRGL1, AtRGL2 and AtRGL3, respectively. Real-time quantitative PCR analysis in the latex showed that, compared with the control, HbGAIP-B transcript levels were significantly down-regulated in latex by consecutive tapping. The transcript levels were significantly up-regulated at 4 hours by gibberellins and ethephon. And methyl jasmonate reduced slightly the expression of HbGAIP-B.

关 键 词：巴西橡胶树 HbGAIP-B 基因克隆 基因表达分析 

分 类 号：S794.1[农业科学—林木遗传育种]