橡胶树5-烯醇式丙酮莽草酸-3-磷酸合成酶基因的克隆及其响应非生物胁迫的表达分析  

Cloning and Expression Analysis of 5-Enolpyruvylshikimate-3-phosphate Synthetase Gene Under Abiotic Stress in Hevea brasiliensis

作　　者：李雅超[1,2,3] 安泽伟[2,3] 谢黎黎[1,2,3] 翟琪麟[1,2,3] 黄华孙[2,3] 

机构地区：[1]海南大学农学院,海南海口570228 [2]国家橡胶树育种中心 [3]中国热带农业科学院橡胶研究所,海南儋州571737

出　　处：《热带作物学报》2013年第5期807-814,共8页Chinese Journal of Tropical Crops

基　　金：国家天然橡胶产业技术体系(No.CARS-34);国家自然科学基金项目(No.30960319;No.31271796);中国热带农业科学院橡胶研究所基本科研业务费专项(No.1630022011002)

摘　　要：5-烯醇式丙酮莽草酸-3-磷酸合成酶(EPSPs)是植物合成芳香族氨基酸的一个重要酶,与许多抗逆相关的次生代谢物的合成有关。本研究利用RACE技术从橡胶树热研7-33-97中获得了全长为2 025 bp的HbEPSPS基因cDNA序列,开放阅读框为1 572 bp,编码523个氨基酸;使用ProtParam tool预测该基因编码的蛋白质分子量为56.01 u,等电点(pI)为7.91。该基因由8个外显子和7个内含子构成,全长4 600 bp。实时荧光定量PCR结果表明,HbEPSPS基因在橡胶树叶片、皮、花、胶乳和胚中均表达,其中以叶片中的表达量为最高。激素、干旱、盐和低温等非生物胁迫都能诱导HbEPSPS基因表达量上调,其中茉莉酸甲酯、乙烯和PEG的上调作用最为显著。橡胶树HbEPSPS基因的克隆及其在非生物胁迫下的表达调控研究将为橡胶树抗逆相关的研究提供理论依据。5-Enolpyruvylshikimate-3-phosphate （EPSPs） synthase is an important enzyme of the shikimate pathway. It is associated with the synthesis of a lot of secondary metabolites related to stress tolerance. In this study, a 2 025 bp HbEPSPS cDNA, containing a 1 572 bp ORF and encoding 523 amino acids, was cloned from CATAS- 73397 by 5＇ and 3＇ RACE. Bioinformatics analysis showed that the molecular weight of HbEPSPS protein was 56.01 u and its isoelectrie point was 7.91. The full length of HbEPSPS gene was 4 600 bp, and there were eight exons and seven introns in HbEPSPS gene. The results of qRT-PCR showed that HbEPSPS could express in leaves, bark, flower, latex and embryo, and among these tissues, but the highest expression level of HbEPSPS was in leaves. The expression of HbEPSPS could be induced by hormones, drought, salt and low temperature, and it was significantly up-regulated under MeJA, ETH and PEG stress, respectively. The cloning and expression analysis of HbEPSPS gene under abiotic stress will provide theoretical basis for rubber tree in the abiotic stress research.

关 键 词：橡胶树 EPSPS 非生物胁迫 莽草酸途径 

分 类 号：S794.1[农业科学—林木遗传育种]