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作物学报 ›› 2012, Vol. 38 ›› Issue (06): 980-987.doi: 10.3724/SP.J.1006.2012.00980

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

水稻稻瘟病菌诱导表达启动子OsQ16p的克隆与功能分析

王光，吴智丹，张磊，刘凤权，邵敏*   

  1. 南京农业大学植物保护学院/农作物生物灾害综合治理教育部重点实验室，江苏南京210095
  • 收稿日期:2011-12-22 修回日期:2012-02-22 出版日期:2012-06-12 网络出版日期:2012-03-29
  • 通讯作者: 邵敏, E-mail: minshao@njau.edu.cn
  • 基金资助:

    本研究由国家高技术研究发展计划(863计划)项目(2007AA10Z188)和国家转基因生物新品种培育项目(2009ZX08001-005B)资助。

Cloning and Functional Analysis of Magnaporthe oryzae-Induced Promoter OsQ16p in Rice

WANG Guang,WU Zhi-Dan,ZHANG Lei,LIU Feng-Quan,SHAO Min*   

  1. College of Plant Protection, Nanjing Agricultural University/Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Nanjing 210095, China
  • Received:2011-12-22 Revised:2012-02-22 Published:2012-06-12 Published online:2012-03-29
  • Contact: 邵敏, E-mail: minshao@njau.edu.cn

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被引次数

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摘要： qRT-PCR分析表明，日本晴OsQ16基因受稻瘟病菌诱导表达。利用PCR技术从日本晴基因组中克隆了该基因编码区5′端上游1 229 bp的启动子序列，命名为OsQ16p。用其取代pBI121中gus基因上游的CaMV35S启动子，构建重组表达载体pBIQ16p，经农杆菌介导转化日本晴，获得转基因植株。GUS组织化学染色和qRT-PCR分析表明: (1) gus基因在抗性愈伤组织和阳性转基因植株中均能表达; (2)转基因植株在接种稻瘟病菌后12 h，GUS表达量是处理前的2.7倍; (3)抗病相关信号分子水杨酸(salicylic acid，SA)和茉莉酸甲酯(methyl jasmonate，MeJA)喷施转基因植株叶面后12 h，GUS表达量分别为处理前的3.1倍和3.5倍。以上结果表明，OsQ16p启动子具有启动活性，并明显受稻瘟病菌、MeJA和SA诱导表达。

关键词: 转基因水稻, 诱导型启动子, OsQ16p, GUS, 稻瘟病菌

Abstract: The qRT-PCR analysis showed that, in Nipponbare (Oryza sativa L. ssp japonica), the expressionof OsQ16 gene was induced by Magnaporthe grisea. The 1 299 bp-fragment of 5'-end of OsQ16 gene, named as OsQ16p,was amplified by PCR from Nipponbare. The plasmids pBIQ16p was constructed by replacing the CaMV35S promoter of pBI121 with the OsQ16p, and transformed into Nipponbare through Agrobacterium-mediated transformation. The analysis of GUS activity and qRT-PCR showed that gus gene could express in transgenic plants and callui. The expression of gus gene in transgenic plants was obviously enhanced by M. grisea. After treatment with the resistance-related signaling molecules SA and MeJA, the GUS activities in transgenic plants were increased by 3.1- and 3.5-fold, respectively. It suggested that M. grisea, SA and MeJA were inductive factors of OsQ16p promoter.

Key words: Transgenic rice, Inducible promoter, OsQ16p, GUS, Magnaporthe grisea

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