橡胶树HbCPA基因的克隆、表达及生物信息学分析  

Cloning,Expression and Bioinformatics Analyses of HbCPA Gene from Hevea brasiliensis

作　　者：杨洪[1] 胡燕玲 岳镒繁 邓治[1] 代龙军[1] 李德军[1] YANG Hong;HU Yan-ling;YUE Yi-fan;DENG Zhi;DAI Long-jun;LI De-jun(Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Biology and Genetic Resources of Rubber Tree for Ministry of Agriculture and Rural Affairs/State Key Laboratory Incubation Base for Cultivation and Physiology for Tropical Crops-Hainan Provincial Key Laboratory of Tropical Crops Cultivation and Physiology,Haikou 571101;College of Agriculture,Nanjing Agricultural University,Nanjing 210095)

机构地区：[1]中国热带农业科学院橡胶研究所农业农村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,海口571101 [2]南京农业大学农学院,南京210095

出　　处：《生物技术通报》2020年第11期39-47,共9页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31570684);中国热带农业科学院基本科研业务费专项资金(1630022020015);国家重点研发计划(2018YFD1000502)。

摘　　要：N-氨甲酰基腐胺酰胺水解酶(N-carbamoylputreseine amidohydrolase,CPA)是高等植物通过精氨酸途径合成腐胺的终反应酶。利用RACE技术从橡胶树中克隆了一个CPA基因,命名为HbCPA。HbCPA序列全长1342 bp,开放阅读框为906 bp,编码301个氨基酸。HbCPA理论分子量约为33.5 kD,理论等电点为6.01。进化分析表明,HbCPA与木薯MeCPA、蓖麻RcCPA和麻风树JcCPA聚为一枝。qRT-PCR结果显示,HbCPA在橡胶树各组织中均有表达,表达量从高到低依次为树皮、叶片、雌花、雄花、茎尖;HbCPA在叶片不同发育时期表达存在变化,淡绿期最高,古铜期和衰老期表达量相当且最低;同健康树树皮相比,HbCPA在死皮树树皮中表达量明显上调;HbCPA表达受乙烯利、茉莉酸甲酯、低温、干旱、高盐、过氧化氢等调控,表明HbCPA可能参与橡胶树产排胶调控及非生物逆境胁迫响应。N-carbamoylputreseine amidohydrolase(CPA)is the final reaction enzyme for higher plants to synthesize putrescine through the arginine pathway.A CPA gene from Hevea brasiliensis was cloned with RACE and RT-PCR and designated as HbCPA.The full length of HbCPA was 1342 bp with an open reading frame of 906 bp and encoded 301 amino acids.The molecular weight and isoelectric point of HbCPA were 33.5 kD and 6.01,respectively.Phylogenetic analyses showed that HbCPA and MeCPA in Manihot esculenta,RcCPA in Ricinus communis and JcCPA in Jatropha curcas clustered into one branch.qRT-PCR analyses demonstrated that HbCPA expressed in each tissue,with the highest expression in barks,followed by leaves,female flowers,male flowers and stem apexes.The expression of HbCPA varied in different developmental stages of leaves,with the highest in the light green period,and the same and lowest expression in the bronze and senescence stages.The expression of HbCPA was significantly up-regulated in barks of tapping panel dryness(TPD)tree compared with the healthy rubber tree.HbCPA expression was also regulated by ethephon,MeJA,low temperature,drought,high salt and hydrogen peroxide.All these results suggest that HbCPA may be involved in the regulation of rubber production and debinding in rubber tree and the response to abiotic stress.

关 键 词：橡胶树 N-氨甲酰基腐胺酰胺水解酶 非生物胁迫 生物信息学 表达分析 

分 类 号：Q943.2[生物学—植物学] S794.1[农业科学—林木遗传育种]