中国水稻科学

中国水稻科学

• 研究报告 • 上一篇    下一篇

水稻OsWRKY89基因启动子的表达特性

郝中娜1;王海华1;郭泽建2，*   

  1. 1浙江大学 生物技术研究所， 浙江 杭州 310029； 2中国农业大学 植物病理系， 北京 100094
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2006-03-10 发布日期:2006-03-10

Expression Characteristics of a Rice OsWRKY89 Gene Promoter

HAO Zhong-na , WANG Hai-hua , GUO Ze-jian   

  • Received:1900-01-01 Revised:1900-01-01 Online:2006-03-10 Published:2006-03-10

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被引次数

7

摘要： 利用PCR技术从粳稻品种秀水11中扩增了转录因子WRKY89编码区5′上游大小为1470 bp的调控序列，命名为OsW89p，将它与GUS基因融合，构建植物表达载体。用农杆菌介导法将载体转化水稻，GUS组织化学分析结果表明，OsW89p驱动的GUS基因在转基因植株各个时期的叶、茎和叶鞘中均有表达；在花器的外稃和花药中有活性而在花粉和成熟种子中均无活性；种子萌发时在胚中有活性；2.5叶期时在种子根、不定根和侧根中均有活性但根尖中无活性，5叶期时根部只有侧根中有活性。分析了GUS在T1代苗期时的诱导表达特性，GUS荧光测定结果表明：1) GUS的表达被茉莉酸甲酯增强，诱导倍数是4.0，被2,4D抑制，水杨酸对OsW89p的表达几乎没有影响；2) 用NaCl和PEG这2种非生物因子处理转基因苗根部，根中GUS的表达被抑制，但叶片中可被诱导增强；3) GUS的表达可被紫外线照射、高温(42℃)、低温(5℃)和伤处理这4种非生物逆境因子增强，其中以紫外线照射处理的诱导倍数最高。

关键词: WRKY基因, 启动子, 表达特性, 水稻

Abstract: A 1470 bp promoter fragment of a rice transcription factor WRKY89 gene(OsW89p) amplified from a rice variety (Oryza sativa L. subsp. japonica cv. Xiushui 11) by polymerase chain reaction(PCR) technique, was fused with GUS gene and transformed into rice embryos by Agrobacterium-mediated method. The results of histochemical GUS staining showed that leaves, culms and sheaths of the transformants had GUS activities at different developmental stages. In terms of flower organ and roots, lemmae and anthers had GUS activities except for pollen, and primary, adventitious and lateral roots had the activities but root tips not at the 2.5-leaf stage, and only lateral roots had the activities at the 5-leaf stage. There were GUS activities in the embryos of the germinating seeds, but not in mature seeds. The inducible expression characteristic of OsW89p in rice detected by GUS fluorometric measurement revealed that:1)Expression level of GUS was increased 4-fold by MeJA treatment, and repressed by the presence of 2,4-D, but was not influenced by salicylic acid; 2)The expression of GUS was repressed in the roots and promoted in the leaves by NaCl and PEG; 3)Ultraviolet, high temperature (42℃), low temperature(5℃) and injury treatments can increase the expression of GUS in leaves, and the induced efficiency of ultraviolet was the highest.

Key words: WRKY gene, promoter, expression characteristic, rice