欢迎访问作物学报,今天是

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 网络出版日期:2017-05-23
  • 通讯作者: 张明才, E-mail: zmc1214@163.com, Tel: 010-62733049
  • 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 Published:2017-09-12 Published online:2017-05-23
  • Contact: Zhang mingcai, E-mail: zmc1214@163.com, Tel: 010-62733049
  • 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

[1] Xia X J, Huang L F, Zhou Y H, Mao W H, Shi K, Wu J X, Asami T, Chen Z, Yu J Q. Brassinosteroids promote photosynthesis and growth by enhancing activation of Rubiseo and expression of photosynthetic genes in Cucumis sativus.Planta, 2009, 230: 1185–1196
[2] Yu J Q, Huang L F, Hu W H, Zhou Y H, Mao W H, Ye S F, Nogués S. A role for brassinosteroids in the regulation of photosynthesis in Cucumis sativus. J Exp Bot, 2004, 55:1135–1143
[3] Vriet C, Russinova E, Reuzeau C. Boosting crop yields with plant steroids. Plant Cell, 2012, 24: 842–857
[4] Yang C J, Zhang C, Lu Y N, Jin J Q, Wang X L. The mechanisms of brassinosteroids’ action: from signal transduction to plant development. Mol Plant, 2011, 4:588–600
[5] Wang Z Y, Bai M Y, Oh E, Zhu J Y. Brassinosteroid signaling network and regulation of photomorphogenesis. Annu Rev Genet, 2012, 46:701–724
[6] Morinaka Y, Sakamoto T, Inukai Y, Agetsuma M, Kitano H, Ashikari M, Matsuoka M. Morphological alteration caused by brassinosteroid insensitivity increases the biomass and grain production of rice. Plant Physiol, 2006, 141: 924–931
[7] Yamamuro C, Ihara Y, Wu X, Noguchi T, Fujioka S, Takatsuto S, Ashikari M, Kitano H, Matsuoka M. Loss of function of a rice brassinosteroid insensitive1 homolog prevents internode elongation and bending of the lamina joint. Plant Cell, 2000, 12: 1591–1605
[8] Nakamura A, Fujioka S, Sunohara H, Kamiya N, Hong Z, Inukai Y, Miura K, Takatsuto S, Yoshida S, Ueguchi-Tanaka M, Hasegawa Y, Kitano H, Matsuoka M. The role of OsBRI1 and its homologous genes, OsBRL1 and OsBRL3, in rice. Plant Physiol, 2006, 140: 580–590
[9] Chono M, Honda I, Zeniya H, Yoneyama K, Saisho D, Takeda K, Takatsuto S, Hoshino T, Watanabe Y. A semidwarf phenotype of barley uzu results from a nucleotide substitution in the gene encoding a putative brassinosteroid receptor. Plant Physiol, 2003, 133: 1209–1219
[10] Clouse S D, Langford M, McMorris T C. A brassinosteroid-insensitive mutant in Arabidopsis thaliana exhibits multiple defects in growth and development. Plant Physiol, 1996, 111: 671–678
[11] Nomura T, Bishop G J, Kaneta T, Reid J B, Chory J, Yokota T. The LKA gene is a BRASSINOSTEROID INSENSITIVE 1 homolog of pea. Plant J, 2003, 36: 291–300
[12] Sun Y, Fokar M, Asami T, Yoshida S, Allen R D. Characterization of the brassinosteroid insensitive 1 genes of cotton. Plant Mol Biol, 2004, 54: 221–232
[13] Liu T S, Zhang J P, Wang M Y, Wang Z Y, Li G F, Qu L, Wang G Y. Expression and functional analysis of ZmDWF4, an ortholog of Arabidopsis DWF4 from maize (Zea mays L.). Plant Cell Rep, 2007, 26: 2091–2099
[14] Tao Y Z, Zheng J, Xu Z M, Zhang X H, Zhang K, Wang G Y. Functional analysis of ZmDWF1, a maize homolog of the Arabidopsis brassinosteroids biosynthetic DWF1/DIM gene. Plant Sci, 2004, 167: 743–751
[15] Makarevitch I, Thompson A, Muehlbauer G J, Springer N M. Brd1gene in maize encodes a brassinosteroid C-6 oxidase. PLoS One, 2012, 7(1): e30798
[16] Kir G, Ye H X, Nelissen H, Neelakandan A K, Kusnandar A S, Luo A D, Inzé D, Sylvester A W, Yin Y H, Becraft P W. RNA interference knockdown of BRASSINOSTEROID INSENSITIVE1 in maize reveals novel functions for brassinosteroid signaling in controlling plant architecture. Plant Physiol, 2015, 169: 826–839
[17] Clough S J, Bent A F. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J, 1998, 16:735–743
[18] Kim T W, Guan S, Burlingame A L, Wang Z Y. The CDG1 kinase mediates brassinosteroid signal transduction from BRI1 receptor kinase to BSU1 phosphatase and GSK3-like kinase BIN2. Mol Cell, 2011, 43:561–571
[19] Noguchi T, Fujioka S, Choe S, Takatsuto S, Yoshida S, Yuan H, Feldmann K A, Tax F E. Brassinosteroid-insensitive dwarf mutants of Arabidopsis accumulate brassinosteroids. Plant Physiol, 1999, 121, 743–752
[20] Clouse S D, Langford M, McMorris T C. A brassinosteroid-insensitive mutant in Arabidopsis thaliana exhibits multiple defects in growth and development. Plant Physiol, 1996, 111: 671–678
[21] Hartwig T, Corvalan C, Best N B, Budka J S, Zhu J Y, Choe S, Schulz B. Propiconazole is a specific and accessible brassinosteroid (BR) biosynthesis inhibitor for arabidopsis and maize. PLoS One, 2012, 7(5): e36625
[22] Wang M, Sun S, Wu C X, Han T F, Wang Q Y. Isolation and characterization of the brassinosteroid receptor gene (GmBRI1) from Glycine max. Int J Mol Sci, 2014, 15:3871–3888
[23] Hu Y, Yu D. BRASSINOSTEROID INSENSITIVE2 interacts with ABSCISIC ACID INSENSITIVE5 to mediate the antagonism of brassinosteroids to abscisic acid during seed germination in arabidopsis. Plant Cell, 2014, 26: 4394–4408
[24] Wu C Y, Trieu A, Radhakrishnan P, Kwok S F, Harris S, Zhang K, Wang J L, Wan J M, Zhai H Q, Takatsuto S, Matsumoto S, FujiokaS, Feldmann K A, Pennell R I. Brassinosteroids regulate grain filling in rice. Plant Cell, 2008, 20: 2130–2145

[1] 肖颖妮, 于永涛, 谢利华, 祁喜涛, 李春艳, 文天祥, 李高科, 胡建广. 基于SNP标记揭示中国鲜食玉米品种的遗传多样性[J]. 作物学报, 2022, 48(6): 1301-1311.
[2] 崔连花, 詹为民, 杨陆浩, 王少瓷, 马文奇, 姜良良, 张艳培, 杨建平, 杨青华. 2个玉米ZmCOP1基因的克隆及其转录丰度对不同光质处理的响应[J]. 作物学报, 2022, 48(6): 1312-1324.
[3] 王丹, 周宝元, 马玮, 葛均筑, 丁在松, 李从锋, 赵明. 长江中游双季玉米种植模式周年气候资源分配与利用特征[J]. 作物学报, 2022, 48(6): 1437-1450.
[4] 杨欢, 周颖, 陈平, 杜青, 郑本川, 蒲甜, 温晶, 杨文钰, 雍太文. 玉米-豆科作物带状间套作对养分吸收利用及产量优势的影响[J]. 作物学报, 2022, 48(6): 1476-1487.
[5] 陈静, 任佰朝, 赵斌, 刘鹏, 张吉旺. 叶面喷施甜菜碱对不同播期夏玉米产量形成及抗氧化能力的调控[J]. 作物学报, 2022, 48(6): 1502-1515.
[6] 徐田军, 张勇, 赵久然, 王荣焕, 吕天放, 刘月娥, 蔡万涛, 刘宏伟, 陈传永, 王元东. 宜机收籽粒玉米品种冠层结构、光合及灌浆脱水特性[J]. 作物学报, 2022, 48(6): 1526-1536.
[7] 单露英, 李俊, 李亮, 张丽, 王颢潜, 高佳琪, 吴刚, 武玉花, 张秀杰. 转基因玉米NK603基体标准物质研制[J]. 作物学报, 2022, 48(5): 1059-1070.
[8] 雷新慧, 万晨茜, 陶金才, 冷佳俊, 吴怡欣, 王家乐, 王鹏科, 杨清华, 冯佰利, 高金锋. 褪黑素与2,4-表油菜素内酯浸种对盐胁迫下荞麦发芽与幼苗生长的促进效应[J]. 作物学报, 2022, 48(5): 1210-1221.
[9] 许静, 高景阳, 李程成, 宋云霞, 董朝沛, 王昭, 李云梦, 栾一凡, 陈甲法, 周子键, 吴建宇. 过表达ZmCIPKHT基因增强植物耐热性[J]. 作物学报, 2022, 48(4): 851-859.
[10] 刘磊, 詹为民, 丁武思, 刘通, 崔连花, 姜良良, 张艳培, 杨建平. 玉米矮化突变体gad39的遗传分析与分子鉴定[J]. 作物学报, 2022, 48(4): 886-895.
[11] 闫宇婷, 宋秋来, 闫超, 刘爽, 张宇辉, 田静芬, 邓钰璇, 马春梅. 连作秸秆还田下玉米氮素积累与氮肥替代效应研究[J]. 作物学报, 2022, 48(4): 962-974.
[12] 徐宁坤, 李冰, 陈晓艳, 魏亚康, 刘子龙, 薛永康, 陈洪宇, 王桂凤. 一个新的玉米Bt2基因突变体的遗传分析和分子鉴定[J]. 作物学报, 2022, 48(3): 572-579.
[13] 宋仕勤, 杨清龙, 王丹, 吕艳杰, 徐文华, 魏雯雯, 刘小丹, 姚凡云, 曹玉军, 王永军, 王立春. 东北主推玉米品种种子形态及贮藏物质与萌发期耐冷性的关系[J]. 作物学报, 2022, 48(3): 726-738.
[14] 渠建洲, 冯文豪, 张兴华, 徐淑兔, 薛吉全. 基于全基因组关联分析解析玉米籽粒大小的遗传结构[J]. 作物学报, 2022, 48(2): 304-319.
[15] 张倩, 韩本高, 张博, 盛开, 李岚涛, 王宜伦. 控失尿素减施及不同配比对夏玉米产量及氮肥效率的影响[J]. 作物学报, 2022, 48(1): 180-192.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!