State Key Laboratory of Cotton Biology, Institute of Cotton Research of Chinese Academy of Agricultural Science, Anyang 455000, China.
College of Biology and Food Engineering, Anyang Institute of Technology, Anyang 455000, China.
Int J Mol Sci. 2020 Sep 27;21(19):7126. doi: 10.3390/ijms21197126.
Verticillium wilt will seriously affect cotton yield and fiber quality. BEL1-Like transcription factors are involved in the regulation of secondary cell wall (SCW) formation, especially the biosynthesis of lignin that also plays a key role in cotton disease resistance. However, there is no report on the role of BEL1-Like transcription factor in the regulation of plant biological stress. In this study, tissue expression pattern analysis showed that a BEL1-Like transcription factor GhBLH7-D06 was predominantly expressed in vascular tissues and the SCW thickening stage of fiber development, while its expression could also respond to infection and the phytohormone MeJA treatment, which indicated that GhBLH7-D06 might be involved in the defense response of Verticillium wilt. Using virus-induced gene silencing (VIGS) technology, we found silencing the expression of could enhance the resistance of cotton plants to Verticillium wilt, and the acquisition of resistance might be mainly due to the significant overexpression of genes related to lignin biosynthesis and JA signaling pathway, which also proves that GhBLH7-D06 negatively regulates the resistance of cotton to Verticillium wilt. Based on the results of yeast two-hybrid (Y2H) library screening and confirmation by bimolecular fluorescence complementary (BiFC) experiment, we found an Ovate Family Protein (OFP) transcription factor GhOFP3-D13 which was also a negative regulator of cotton Verticillium wilt resistance could that interacts with GhBLH7-D06. Furthermore, the dual-luciferase reporter assay and yeast one-hybrid (Y1H) experiment indicated that GhBLH7-D06 could target binding to the promoter region of to suppress its expression and eventually lead to the inhibition of lignin biosynthesis. In general, the GhBLH7-D06/GhOFP3-D13 complex can negatively regulate resistance to Verticillium wilt of cotton by inhibiting lignin biosynthesis and JA signaling pathway.
黄萎病会严重影响棉花的产量和纤维质量。BEL1 样转录因子参与次生细胞壁(SCW)形成的调节,特别是木质素的生物合成,木质素在棉花抗病性中也起着关键作用。然而,目前还没有关于 BEL1 样转录因子在植物生物胁迫调节中的作用的报道。在这项研究中,组织表达模式分析表明,BEL1 样转录因子 GhBLH7-D06 在维管束组织和纤维发育的次生细胞壁增厚阶段表达量较高,而其表达也能响应 感染和植物激素 MeJA 处理,这表明 GhBLH7-D06 可能参与了黄萎病的防御反应。利用病毒诱导的基因沉默(VIGS)技术,我们发现沉默 的表达可以增强棉花植株对黄萎病的抗性,而获得抗性可能主要是由于与木质素生物合成和 JA 信号通路相关的基因的显著过表达,这也证明了 GhBLH7-D06 负调控棉花对黄萎病的抗性。基于酵母双杂交(Y2H)文库筛选的结果和双分子荧光互补(BiFC)实验的验证,我们发现 Ovate Family Protein(OFP)转录因子 GhOFP3-D13 也是棉花黄萎病抗性的负调控因子,它可以与 GhBLH7-D06 相互作用。此外,双荧光素酶报告基因检测和酵母单杂交(Y1H)实验表明,GhBLH7-D06 可以靶向结合 启动子区域抑制其表达,最终导致木质素生物合成的抑制。总的来说,GhBLH7-D06/GhOFP3-D13 复合物可以通过抑制木质素生物合成和 JA 信号通路来负调控棉花对黄萎病的抗性。