橡胶树胶乳HbPLDα1基因及其启动子的克隆与生物信息学分析

Cloning and Bioinformatical Analysis of Phospholipase Dα1 Gene and Its Promoter in the Latex of Rubber Tree(Hevea brasiliensis Muell.Arg.)

植物磷脂酶PLDα1与伤害信号转导密切相关,是伤害诱导内源茉莉酸(Jasmonic acid,JA)生物合成的关键酶之一。橡胶树胶乳PLDα1基因(HbPLDα1)表达的研究将有助于揭示橡胶树乳管细胞JA信号转导及其调控橡胶生物合成的机制。在EST序列的基础上,通过RACE和Genome Walking方法分别克隆了橡胶树胶乳的HbPLDα1基因及其启动子序列。HbPLDα1基因的cDNA全长为2 870 bp,包含长度为2 427 bp的完整开放阅读框(ORF),具有典型的植物PLDα蛋白保守功能域,与同属大戟科的蓖麻和麻风树的PLDα1基因亲缘关系最近。HbPLDα1基因启动子区域长为1 559 bp,除含有TATA box和CAAT box等基本顺式作用元件外,还存在JA和脱落酸等激素响应元件以及干旱胁迫等环境信号响应元件,这表明HbPLDα1基因的表达可能受激素和环境信号的调控,在橡胶树乳管细胞对激素和环境信号的响应过程中发挥重要作用。

Phospholipase D alpha 1（PLDα1） is involved in wound signal transduction in plant cells,and is a key component for wound-induced jasmonic acid（JA） production.The investigation of PLDα1 expression would facilitate unraveling the signaling pathway of JA and its regulation of the rubber biosynthesis in Hevea brasiliensis（rubber tree）.The PLDα1 gene from the Hevea latex,HbPLDα1,and its promoter were cloned based on the EST sequences using RACE and Genome Walking.The full-length cDNA of HbPLDα1 gene was 2 870 bp with a complete ORF of 2 427 bp.HbPLDα1 had the similar typical conserved domains of plant PLD with those of other plants,and showed the highest identity with those of PLDα1 from Ricinus communis and Jatropha curcas.The promoter of HbPLDα1 was 1 559 bp long,including not only the ordinary cis-acting elements of TATA and CAAT box,but also such cis-acting elements responsive to phytohormones（JA,ABA,GB） and environmental stresses.The data demonstrated that the expressed of HbPLDα1 gene could be regulated by signals of both plant hormones and environmental stresses,and it’s suggested that HbPLDα1 might play key roles in the responses to hormone stimulation and environmental stresses for rubber trees.