麻风树JcGASA6基因启动子克隆及其功能初步分析  

Cloning and preliminary function analysis of Jatropha curcas JcGASA6 gene promoter

作　　者：刘利伟 范付华 徐刚 LIU Liwei;FAN Fuhua;XU Gang(Institute for Forest Resources and Environment of Guizhou,Key Laboratory of Forest Cultivation in Plateau Mountain of GuizhouProvince,College of Forestry,GuizhouUniversity,Guiyang 550025,China)

机构地区：[1]贵州大学贵州省森林资源与环境研究中心/贵州省高原山地林木培育重点实验室/林学院,贵阳550025

出　　处：《植物生理学报》2023年第7期1321-1328,共8页Plant Physiology Journal

基　　金：国家自然科学基金(31760198)。

摘　　要：JcGASA6基因介导了赤霉素(GA)等激素对麻风树(Jatropha curcas)成花发育的调控过程。为了明确JcGASA6基因启动子中响应水杨酸(SA)、赤霉素和脱落酸(ABA)的元件所在位置,克隆了JcGASA6基因上游2063 bp启动子序列。通过PlantCARE预测出启动子区域内存在多个激素响应作用元件、光响应作用元件以及参与缺氧特异性诱导的增强子样元件等。对启动子进行5′端缺失克隆,将长度分别为1434、1365、1225、473、130 bp的启动子克隆入以GUS基因为报告基因的载体,进行本氏烟草(Nicotiana benthamiana)瞬时转化,发现JcGASA6启动子的核心元件位置处于-473~-130 bp之间。水杨酸(100μmol·L^(-1))、脱落酸(100μmol·L^(-1))和赤霉素(100μmol·L^(-1))处理下,定量分析JcGASA6启动子瞬时转化烟草叶片的GUS活性,发现水杨酸、脱落酸和赤霉素都抑制启动子的活性。推测启动子-1225~-130 bp存在负调控的SA响应元件,-1365~-130 bp存在负调控的GA响应元件,-2063~-1434和-1225~-473 bp区间内存在负调控的ABA响应元件。这些结果为深入研究JcGASA6基因在麻风树花发育过程中的调控机制提供参考。JcGASA6 gene mediates the regulation process of gibberellin and other hormones on the flowering development of Jatropha curcas.In order to clarify the location of the elements responding to salicylic acid(SA),gibberellin and abscisic acid(ABA)in the promoter of JcGASA6 gene,the 2063 bp promoter sequence upstream of JcGASA6 gene was cloned.PlantCARE predicted that there are multiple hormone response elements,light response elements and enhancer-like elements involved in hypoxia-specific induction in the promoter region.The 5'-terminal deletion of the promoter was cloned,and the promoter lengths of 1434,1365,1225,473 and 130 bp were cloned into the GUS reporter vector for tobacco(Nicotiana benthamiana)transient transformation.The results show that the core element of JcGASA6 promoter is located between-473 and-130 bp.Under the treatment of salicylic acid(SA;100μmol-L^(-1)),abscisic acid(ABA;100μmol-L^(-1))and gibberellin(GAs;100μmol-L^(-1)),the GUS activity of JcGASA6 promoter transiently transformed tobacco leaves was quantitatively analyzed.It was found that SA,ABA and GAg inhibited the promoter activity.It is speculated that the promoter-1225~-130 bp has a negatively regulatory SA response element,-1365~-130 bp has a negatively regulatory GA response element,and-2063~-1434 and-1225~-473 bp have negatively regulatory ABA response elements.These results provide a reference for further study of the regulatory mechanism of JcGASA6 gene in the development of J.curcas flowers.

关 键 词：麻风树 JcGASA6启动子 瞬时表达 GUS活性 

分 类 号：S794.9[农业科学—林木遗传育种]