Zhong Chenli, Lan Hujiao, Wang Wenxu, Zhao Yating, Ma Xiaohan, Liu Jianzhong
College of Life Sciences, Zhejiang Normal University, Jinhua 321004, Zhejiang, China.
Zhejiang Provincial Key Laboratory of Biotechnology on Specialty Economic Plants, College of Life Sciences, Zhejiang Normal University, Jinhua 321004, Zhejiang, China.
Sheng Wu Gong Cheng Xue Bao. 2024 Oct 25;40(10):3810-3822. doi: 10.13345/j.cjb.230805.
The WRKY transcription factor gene family is a plant-specific transcription factor that plays important roles defense responses. Studies in model plant demonstrated that WRKYs function downstream of mitogen activated-protein kinase (MAPK) signaling cascade and participate in defense responses through activating the expression of defense-related genes. However, the roles of WRKYs in defense responses have not been previously investigated in paleopolyploidy soybean. Bioinfomatic analysis revealed that there are three pair of genes in the soybean genome. The identity of first two pair of GmWRKY33 genes is greater than 84% (named as ). The identity of genes within the same pair is greater than 95%. A 300 bp fragment highly homologous to these four was chosen to clone into bean pod mosaic virus (BPMV)-based silencing vector (BPMV-VIGS) to achieve the goal of silencing four genes simultaneously. In this study, we simultaneously silenced four homologous genes of using a bean pod mottle virus (BPMV) vector carrying a single fragment of . Comparing the silenced plants with the vector control plants, no evident morphological phenotypes were observed. However, the silenced plants exhibited significantly reduced resistance to pv. (), pv. (), as well as to soybean mosaic virus (SMV). Furthermore, we demonstrated that silencing these genes significantly inhibited the activation of MPK3/MPK6 induced by infection. Collectively, our results suggest that WRKY33As are involved in soybean immunity through regulating the transcription of genes or activating the kinase activities of MPK3/6. Taken together, our results demonstrated that WRKY33As are positive regulators of soybean immune responses.
WRKY转录因子基因家族是植物特有的转录因子，在防御反应中发挥重要作用。在模式植物中的研究表明，WRKY蛋白在丝裂原活化蛋白激酶（MAPK）信号级联反应的下游发挥作用，并通过激活防御相关基因的表达参与防御反应。然而，此前尚未在古多倍体大豆中研究WRKY蛋白在防御反应中的作用。生物信息学分析表明，大豆基因组中有三对基因。前两对GmWRKY33基因的同一性大于84%（命名为 ）。同一对基因内的同一性大于95%。选择与这四个基因高度同源的300 bp片段克隆到基于菜豆荚斑驳病毒（BPMV）的沉默载体（BPMV-VIGS）中，以实现同时沉默四个基因的目的。在本研究中，我们使用携带单个片段的菜豆斑驳病毒（BPMV）载体同时沉默了四个同源基因。将沉默植株与载体对照植株进行比较，未观察到明显的形态表型。然而，沉默植株对大豆疫霉（ ）、番茄疫霉（ ）以及大豆花叶病毒（SMV）的抗性显著降低。此外，我们证明沉默这些基因显著抑制了由感染诱导的MPK3/MPK6的激活。总体而言，我们的结果表明WRKY33A通过调节基因转录或激活MPK3/6的激酶活性参与大豆免疫。综上所述，我们的结果表明WRKY33A是大豆免疫反应的正调控因子。
