Gao Ying, Jia Shuangwei, Wang Chunlian, Wang Fujun, Wang Fajun, Zhao Kaijun
National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI), Institute of Crop Science, Chinese Academy of Agriculture Sciences (CAAS), Beijing 100081, China.
National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI), Institute of Crop Science, Chinese Academy of Agriculture Sciences (CAAS), Beijing 100081, China
J Exp Bot. 2016 Aug;67(15):4647-58. doi: 10.1093/jxb/erw240. Epub 2016 Jun 27.
We previously identified the W-box-like-4 (Wbl-4) element (GTAGTGACTCAT), one of six Wbl elements in the BjC-P promoter of the unusual chitinase gene BjCHI1 from Brassica juncea, as the core element responsive to fungal infection. Here, we report the isolation and characterization of the cognate transcription factor interacting with the Wbl-4 element. Using Wbl-4 as a target, we performed yeast one-hybrid screening of a B. juncea cDNA library and isolated an R2R3-MYB transcription factor designated as BjMYB1. BjMYB1 was localized in the nucleus of plant cells. EMSA assays confirmed that BjMYB1 binds to the Wbl-4 element. Transiently expressed BjMYB1 up-regulated the activity of the BjC-P promoter through its binding to the Wbl-4 element in tobacco (Nicotiana benthamiana) leaves. In B. juncea, BjMYB1 displayed a similar induced expression pattern as that of BjCHI1 upon infection by the fungus Botrytis cinerea Moreover, heterogeneous overexpression of BjMYB1 significantly elevated the resistance of transgenic Arabidopsis thaliana to the fungus B. cinerea These results suggest that BjMYB1 is potentially involved in host defence against fungal attack through activating the expression of BjCHI1 by binding to the Wbl-4 element in the BjC-P promoter. This finding demonstrates a novel DNA target of plant MYB transcription factors.
我们之前鉴定出了W-box样-4(Wbl-4)元件(GTAGTGACTCAT),它是来自芥菜型油菜的异常几丁质酶基因BjCHI1的BjC-P启动子中六个Wbl元件之一,是对真菌感染作出响应的核心元件。在此,我们报告了与Wbl-4元件相互作用的同源转录因子的分离与特性分析。以Wbl-4为靶点,我们对芥菜型油菜cDNA文库进行了酵母单杂交筛选,并分离出一个命名为BjMYB1的R2R3-MYB转录因子。BjMYB1定位于植物细胞的细胞核中。电泳迁移率变动分析(EMSA)证实BjMYB1与Wbl-4元件结合。瞬时表达的BjMYB1通过其与烟草(本氏烟草)叶片中Wbl-4元件的结合上调了BjC-P启动子的活性。在芥菜型油菜中,灰葡萄孢菌感染后,BjMYB1呈现出与BjCHI1相似的诱导表达模式。此外,BjMYB1的异源过表达显著提高了转基因拟南芥对灰葡萄孢菌的抗性。这些结果表明,BjMYB1可能通过与BjC-P启动子中的Wbl-4元件结合来激活BjCHI1的表达,从而参与宿主对真菌攻击的防御。这一发现揭示了植物MYB转录因子的一个新的DNA靶点。