玉米脱氧麦根酸分泌通道蛋白基因YS3启动子的克隆与启动活性分析

发布时间：2018-02-14 07:22

本文关键词： 玉米脱氧麦根酸 YS基因启动子 GUS基因　出处：《遗传》2016年06期 　论文类型：期刊论文

【摘要】：缺铁是世界范围内农业生产面临的严重问题,玉米通过分泌脱氧麦根酸(2’-deoxymugineic acid,DMA)吸收利用土壤中的难溶性铁。为探明玉米DMA分泌通道蛋白基因YS3的表达和调控机制,本文通过克隆获得长为2813 bp的YS3基因启动子,该序列含有大量TATA-box、CAAT-box等启动子基本元件,以及光响应、激素调控等多个顺式调控元件;构建YS3启动子驱动GUS基因的植物表达重组载体p CAMBIA-YS3GUS,利用农杆菌介导转化拟南芥,获得p YS3::GUS转基因植株,对转基因植株进行GUS组织化学染色,并通过石蜡切片技术对转基因植株进行组织观察,分析p YS3::GUS转基因植株中YS3基因启动子的活性。结果表明,YS3启动子主要驱动GUS基因在拟南芥根部表达,且主要集中在根部表皮细胞,机械损伤可激发YS3启动子活性,驱动GUS基因在损伤临近部位表达。本研究对于理解玉米DMA分泌的分子调控机理具有重要意义。
[Abstract]:Iron deficiency is a serious problem in agricultural production all over the world. Maize absorbs and uses insoluble iron in soil by secreting deoxymugineic acid DMAs. In order to investigate the expression and regulation mechanism of DMA secretory channel protein gene YS3 in maize, In this paper, a 2813 BP YS3 promoter was cloned, which contains a large number of basic promoter elements such as TATA-box CAAT-box, photo-response, hormone regulation and many other cis-regulatory elements. The plant expression vector pCAMBIA-YS3GUSwas constructed by YS3 promoter, and transformed into Arabidopsis thaliana by Agrobacterium tumefaciens. The transgenic plants were obtained by GUS histochemical staining. The tissue of transgenic plants was observed by paraffin section technique, and the activity of YS3 gene promoter in p YS3::GUS transgenic plants was analyzed. The results showed that YS3 promoter mainly drives the expression of GUS gene in Arabidopsis thaliana root. Mechanical injury can stimulate the activity of YS3 promoter and drive the expression of GUS gene in the vicinity of injury. This study is of great significance in understanding the molecular regulation mechanism of DMA secretion in maize.
【作者单位】：山东农业大学农学院作物生物学国家重点实验室;
【基金】：国家自然科学基金项目(编号:31101160,31271808) 山东省现代农业产业技术体系创新团队岗位专家项目(编号:SDAIT-01-022-02) 山东省博士后创新项目专项资金(编号:201102038)资助~~
【分类号】：Q943.2;S513

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