巴西橡胶树HbMCS1基因的克隆及表达分析  

Cloning of HbMCS1 Gene from Hevea brasiliensis and Its Expression Analysis

作　　者：雷美玉[1,2] 李辉亮[1] 彭世清[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所,海口571101 [2]海南大学农学院,海口570228

出　　处：《农业生物技术学报》2009年第3期482-487,共6页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.3056012和30760203);中央级公益性科研院所基本科研业务费专项资金项目(No.ITBBZD0712)资助

摘　　要：2C-甲基-D-赤藓糖醇-2,4-环化磷酸合成酶(2C-methyl-D-erythritol2,4-cyclodiphosphate synthase,MCS)是异戊烯基焦磷酸合成途径之一,甲基赤藓糖磷酸(methylerythritol phosphate,MEP)途径中的第5个酶,催化2-磷酸-4-(胞苷-5'-二磷酸)-2-C-甲基-D-赤藓糖醇生成2-C-甲基-D-赤藓糖醇-2-4-环化磷酸。根据植物MCS的同源序列设计引物,通过RT-PCR结合RACE的方法在橡胶树(Hevea brasiliensis)中获得了与其相应的MCS基因,命名为HbMCS1(GenBank登录号:FJ196164)。序列分析表明HbMCS1长965bp,编码241个氨基酸,该氨基酸序列与长春花(Catharanthus roseus)、拟南芥(Arabidopsis thaliana)、水稻(Oryzasativa)、银杏(Ginkgo biloba)和三尖杉(Cephalotaxus fortunei)的MCS同源性分别达到70.8%、69.4%、64.9%和63.3%和62.2%。半定量RT-PCR结果显示,割胶诱导胶乳HbMCS1的表达,乙烯对HbMCS1的表达几乎没有影响;HbMCS1的表达具有组织差异性,在愈伤组织中大量表达,在叶片和胶乳中微量表达。2C-methyl-D-erythritol 2, 4-cyclodiphosphate synthase （MCS） is the fifth enzyme of the nonmevalonate terpenoid pathway for isopentenyl diphosphate biosynthesis and is involved in the methylerythritol phosphate （MEP） pathway. A MCS gene was isolated from Hevea brasiliensis by RT-PCR and RACE based on the conserved sequences of plant MCS genes. Sequence analysis showed that the cDNA, which is designated HbMCS1, consisted of 965 bp and encoded a protein of 241 amino acids with a predicted molecular mass of 25.82 kD （GenBank accession No. FJ196164）. In comparison, this putative HbMCS1 protein showed 70.8%, 69.4%, 64.9%, 63.3% and 62.2% identities with MCS from Catharanthus roseus, Arabidopsis thaliana, Oryza sativa, Ginkgo biloba and Ccphalotaxus fortunei. Semi-quantitative RT-PCR analysis indicated that the HbMCS1 was more highly expressed in callius than that in latex and leaves. The transcription of HbMCS1 in latex from tapping tree was induced, whereas ethylene had little effect on its expression.

关 键 词：巴西橡胶 HbMCS1 甲基赤藓糖磷酸途径 伤害 

分 类 号：S188[农业科学—农业基础科学]