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作物学报 ›› 2017, Vol. 43 ›› Issue (09): 1261-1271.doi: 10.3724/SP.J.1006.2017.01261

• 作物遗传育种·种质资源·分子遗传学 •    下一篇

玉米ZmBRI1基因的克隆、表达及功能分析

郝岭,张钰石,段留生,张明才*,李召虎   

  1. 植物生长调节剂教育部工程研究中心/中国农业大学农学与生物技术学院农学系, 北京100193
  • 收稿日期:2017-02-23 修回日期:2017-05-10 出版日期:2017-09-12 发布日期:2017-05-23
  • 通讯作者: 张明才, E-mail: zmc1214@163.com, Tel: 010-62733049 E-mail:hao_ling2011@163.com
  • 基金资助:

    本研究由引进国际先进农业科学技术计划(948计划)项目(2011-G19)资助。

Cloning, Expression and Functional Analysis of Brassinosteroid Receptor Gene (ZmBRI1) from Zea MaysL.

HAO Ling,ZHANG Yu-Shi,DUAN Liu-Sheng,ZHANG Ming-Cai*,LI Zhao-Hu   

  1. Engineering Research Center of Plant Growth Regulator, Ministry of Education, College of Agronomy and Biotechnology, China Agricultural University, Department of Agronomy, Beijing 100193, China
  • Received:2017-02-23 Revised:2017-05-10 Online:2017-09-12 Published:2017-05-23
  • Contact: Zhang mingcai, E-mail: zmc1214@163.com, Tel: 010-62733049 E-mail:hao_ling2011@163.com
  • Supported by:

    This study was supported by the Introduction of International Advanced Agricultural Science and Technology Program (948 Program, 2011-G19).

摘要:

油菜素内酯(BRs)是一种重要的甾族类激素,在植物的生长过程中起着重要的作用。本研究利用同源克隆的方法,从玉米B73自交系中获得了一个新的油菜素内酯受体蛋白编码基因ZmBRI1,该基因全长为3369bp,编码1122个氨基酸。亚细胞定位分析表明,ZmBRI1蛋白定位于细胞膜上,而且ZmBRI1在各个玉米的组织器官中都有表达,其中在幼嫩的组织中表达最高。利用转基因技术将ZmBRI1导入BR不敏感突变体bri1-5中,获得的转基因植株修复了其表型,特别是植株高度、叶片形态和果荚大小。与突变体bri1-5比较,油菜素内酯(brassinolide, BL)处理显著抑制转基因株系根系生长;丙环唑(propiconazole, Pcz)处理显著抑制了下胚轴生长;同时转基因株系DWF4CPD基因的表达量下降。此外,在野生型(Ws)中过表达ZmBRI1,过表达ZmBRI1株系显著提高了在ABA抑制条件下的种子萌发率和植株生长,而且下调了ABA响应基因RD29ARD29BABI5RAB18的表达。因此,ZmBRI1不仅参与了植物的形态建成和BR的信号传导,而且参与调控了植物对ABA信号的响应。

关键词: 玉米, ZmBRI1, 转基因植株, 油菜素内酯, ABA

Abstract:

Brassinosteroids (BRs) isone of very important plant steroidal hormones that are essential in a wide variety of physiological processes. In this study, an encoding brassinosteroid receptorhomologous gene was cloned by homology cloning from maizeB73 inbred lines, and designated as ZmBRI1. Sequence analysis revealed that the full length of ZmBRI1 was 3369bp, encoding 1122 amino acids. Moreover,ZmBRI1 protein was localized in cell membrane by the protein subcellular localization analysis and a ubiquitously expressed receptor kinase expressed highly in young tissues. The transformation ofZmBRI1 into the Arabidopsis dwarf mutant bri1-5 restored the phenotype, including plant height, leaf morphology and pod size. Compared to bri1-5, Brassinolide (BL) inhibited significantly the root growth of transgenic lines,and Propiconazole(Pcz) inhibited the hypocotyl growth, and the expression levels of DWF4 and CPD were decreased in the transgenic plants. Furthermore, with ABA treatment, overexpression of ZmBRI1 in wild type increased the germination rate and plant growth, and decreased the expression of ABA downstream genes RD29A, RD29B, ABI5,and RAB18 compared to wild type. Therefore, ZmBRI1 was not only involved in plant morphogenesis and BR signal transduction, but also played a pivotal role in response to ABA signal.

Key words: Maize, ZmBRI1, Transgenic plant, Brassinosteroid, ABA

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