遗传 ›› 2016, Vol. 38 ›› Issue (6): 560-568.doi: 10.16288/j.yczz.15-174

• 研究报告 • 上一篇    下一篇

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

尹朝华, 李岩, 张春庆, 杨翠翠, 吴承来, 刘翔攀, 王明明   

  1. 山东农业大学农学院,作物生物学国家重点实验室,泰安 271018
  • 收稿日期:2015-12-31 修回日期:2016-03-25 出版日期:2016-06-20 发布日期:2016-04-25
  • 通讯作者: 李岩,副教授,硕士生导师,研究方向:作物遗传育种。E-mail: liyan5011@163.com
    张春庆,教授,博士生导师,研究方向:作物遗传育种。E-mail: cqzhang@sdau.edu.cn E-mail:yinzhaohua2009@163.com
  • 作者简介:尹朝华,硕士研究生,专业方向:植物生物技术及其在育种上的应用。E-mail: yinzhaohua2009@163.com
  • 基金资助:
    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)和山东省博士后创新项目专项资金(编号:201102038)资助[Supported by the National Natural Science Foundation of China (Nos; 31101160, 31271808), Shandong Province Modern Agriculture Industrial Production Technology System (No; SDAIT-01-022-02), and Shandong Postdoctoral Innovation Foundation (No; 201102038)]

Cloning and promoter activity analyses of the promoter of 2’-deoxymugineic acid (DMA) secretion channel gene YS3 in maize

Zhaohua Yin, Yan Li, Chunqing Zhang, Cuicui Yang, Chenglai Wu, Xiangpan Liu, Mingming Wang   

  1. State Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural University, Tai’an 271018, China
  • Received:2015-12-31 Revised:2016-03-25 Online:2016-06-20 Published:2016-04-25
  • Supported by:
    Supported by the National Natural Science Foundation of China (Nos; 31101160, 31271808), Shandong Province Modern Agriculture Industrial Production Technology System (No; SDAIT-01-022-02), and Shandong Postdoctoral Innovation Foundation (No; 201102038)

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pCAMBIA-YS3GUS,利用农杆菌介导转化拟南芥,获得pYS3::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pYS3::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方法od3 gmaigensuan有重要意义。

关键词: 玉米, 脱氧麦根酸, YS3基因, 启动子, GUS基因

Abstract: Iron (Fe) deficiency is a world-wide serious agricultural problem. Maize secretes 2’-deoxymugineic acid (DMA) to uptake and utilize Fe from the soil. In order to explore the gene expression patterns of the DMA secretion channel gene YS3, we cloned the 2813 bp YS3 promoter, and constructed the plant expression vector pCAMBIA-YS3GUS. The promoter contains a lot of TATA-boxes and CAAT-boxes, and cis-acting regulatory elements such as the light responsive elements and the hormone responsive elements. Arabidopsis was transformed via Agrobacterium tumefacients-mediated procedures to obtain the pYS3::GUS transgenic plants, which were confirmed by GUS staining. Then, the stained tissue was observed using paraffin section methods and the YS3 promoter activity was also analyzed. We found that the promoter could drive GUS gene expression specifically in the root epidermal cells. Mechanical damage could activate the promoter, and drive the GUS gene expression adjacent to the damage sites. Our results provide a molecular basis to understand the DMA secretion process in maize.

Key words: maize, 2’-deoxymugineic acid, YS3, promoter, GUS