Identification of fungus-responsive cis-acting element in the promoter of Brassica juncea chitinase gene, BjCHI1
Identification of fungus-responsive cis-acting element in the promoter of Brassica juncea chitinase gene, BjCHI1
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DOI：
10.1016/j.plantsci.2013.11.008
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发表时间：
2014-02-01
期刊：
影响因子：
5.2
通讯作者：
Zhao, Kai-Jun
中科院分区：
文献类型：
--
作者：
Gao, Ying;Zan, Xin-Li;Zhao, Kai-Jun
几丁质酶是一组与病程相关的蛋白质。芥菜几丁质酶基因BjCHI 1可被病原真菌高度诱导，表明BjCHI 1的启动子中可能含有响应真菌攻击的特异性顺式作用元件。为了鉴定BjCHI 1启动子（BjC-P）中的真菌应答元件，将BjC-P及其缺失衍生物与GUS报告基因融合，构建了一系列植物转化载体。利用瞬时基因表达系统，系统地检测了GUS报告基因在本氏烟草（Nicotiana benthamiana）叶片中的表达，以及在转基因拟南芥（Arabidopsis）中的表达。组织化学和GUS定量分析表明，W-boxlike元件（GTAGTGACTCAT）在该区域（-668到-657）是真菌反应所必需的，尽管BjC-P中还有另外5个W-box样元件。功能获得性分析表明，（-409至-337）偶联到W-box样元件是真菌诱导的全部幅度所需要的。这些结果揭示了W-box类元件参与植物抗病性调控的新机制，为BjC-P在工程作物中的应用奠定了基础。(C)2013爱思唯尔爱尔兰有限公司版权所有。
Chitinases are a group of pathogenesis-related proteins. The Brassica juncea chitinase gene BjCHI1 is highly inducible by pathogenic fungal infection, suggesting that the promoter of BjCHI1 might contain specific cis-acting element responsive to fungal attack. To identify the fungus-responsive element in BjCHI1 promoter (BjC-P), a series of binary plant transformation vectors were constructed by fusing the BjC-P or its deletion-derivatives to p-glucuronidase (GUS) reporter gene. Expression of the GUS reporter gene was systematically assayed by a transient gene expression system in Nicotiana benthamiana leaves treated with fungal elicitor Hexa-N-Acetyl-Chitohexaose, as well as in transgenic Arabidopsis plants inoculated with fungus Botrytis cinerea. The histochemical and quantitative GUS assays showed that the W-boxlike element (GTAGTGACTCAT) in the region (-668 to -657) was necessary for the fungus-response, although there were another five W-box-like elements in BjC-P. In addition, gain-of-function analysis demonstrated that the fragment (-409 to -337) coupled to the W-box-like element was needed for full magnitude of the fungal induction. These results revealed the existence of a novel regulation mechanism of W-box-like element involved in plant pathogenic resistance, and will benefit the potential application of BjC-P in engineering crops. (C) 2013 Elsevier Ireland Ltd. All rights reserved.