橡胶树DELLA蛋白编码基因HbGAI的克隆与表达分析  

Cloning and Expression Analysis of HbGAI Gene in Rubber Tree(Hevea brasiliensis Muell.Arg.)

作　　者：吴绍华[1] 张世鑫[1] 陈月异[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

出　　处：《西北植物学报》2015年第11期2157-2163,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家天然橡胶产业技术体系(CARS-34-GW1);国家自然科学基金(31300504)

摘　　要：该研究采用RT-PCR与RACE技术,从橡胶树‘热研7-33-97’胶乳中克隆了1个DELLA蛋白编码基因HbGAI(GenBank登录号为KT696439)。HbGAI全长cDNA序列2 050bp,包含1个长1 842bp的完整开放阅读框。序列分析显示,HbGAI基因编码613个氨基酸,其推导的蛋白含有DELLA和GRAS结构域,分子量为66.476kD,理论等电点为5.19,无跨膜结构域,属于亲水性蛋白。进化树分析表明,HbGAI蛋白与其他植物中DELLA蛋白具有较高的相似性,与麻疯树JcGAI和蓖麻RcGAI亲缘关系较近。荧光定量PCR结果显示,割胶和茉莉酸甲酯处理下调胶乳中HbGAI基因的表达,乙烯利处理4h内显著上调胶乳中HbGAI基因的表达,表明HbGAI基因可能在橡胶树割胶、茉莉酸、乙烯响应中发挥作用。In this study,one DELLA protein-coding gene HbGAI（GenBank accession No. KT696439） was obtained from latex of rubber tree clone ＇CATAS 7-33-97’ by using rapid-amplification of cDNA ends （RACE） and RT-PCR. The full-length cDNA of HbGAI is 2 050 bp with a 1 842 bp open reading frame （ORF） ,encoding a deduced polypeptide of 613 amino acids with predicted molecular mass of 66. 476 kD and theoretical isoelectric point of 5.19. It is deduced that HbGAI is a hydrophilic protein which contains the DELLA and GRAS domain, while without the trans-membrane domain. Phylogenetic analysis indicated that HbGAI was closely related with JcGAI and RcGAI. Real-time PCR analysis showed that HbGAI gene ex- pression in latex was down-regulated significantly by consecutive tapping and methyl jasmonate （MeJA） treatment. However, ethephon（an ethylene releaser） treatment enhanced the expression of HbGAI within 4 hours significantly. The results suggested that HbGAI might play key roles in responses of tapping,jas-monic acid and ethylene in rubber tree.

关 键 词：巴西橡胶树 HbGAI 基因克隆 基因表达分析 

分 类 号：Q785[生物学—分子生物学] Q786