Zhang Jun, Cui Weiye, Abdul Haseeb Hafiz, Guo Wei
Key Laboratory of Agro-products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Beijing, China.
Environ Microbiol. 2020 Dec;22(12):5387-5401. doi: 10.1111/1462-2920.15268. Epub 2020 Oct 17.
Previous studies have reported the ability of fungi to overwinter in soil or on crop debris under different environmental conditions, but how fungi adapt to chilling is still largely unknown. In this study, we have identified and characterized the RNA binding protein (RBP) (VdNop12) by screening an Agrobacterium tumefaciens-mediated transformation-mediated insertional mutational library of Verticillium dahliae. We determined that this protein was essential to the pathogen for virulence on cotton plants. VdNop12 contains two tandem RNA recognition motif domains, and its orthologs are widely distributed in filamentous fungi. Mutants produced by disruption of VdNop12 showed defects in vegetative growth, conidiation and cell wall integrity. The mutant also showed an increase in sensitivity to low temperature, as compared to the wildtype and complementation strains. Yeast complementation assay showed that VdNop12 could functionally restore the growth phenotype of ΔScNop12 mutant of Saccharomyces cerevisiae at 15°C. We demonstrated that the VdNop12 is localized in the nucleus, and its loss resulted in the downregulated expression of several genes related to cAMP-PKA and MAPK pathways in V. dahliae. Our results demonstrated a crucial role of RBPs in the regulation of morphology, cold adaption, and pathogenic development in V. dahliae.
以往的研究报道了真菌在不同环境条件下能够在土壤或作物残体上过冬,但真菌如何适应低温仍 largely 未知。在本研究中,我们通过筛选根癌农杆菌介导的大丽轮枝菌转化介导的插入突变文库,鉴定并表征了 RNA 结合蛋白(RBP)(VdNop12)。我们确定该蛋白对病原菌在棉花植株上的毒力至关重要。VdNop12 包含两个串联的 RNA 识别基序结构域,其直系同源物广泛分布于丝状真菌中。通过破坏 VdNop12 产生的突变体在营养生长、产孢和细胞壁完整性方面表现出缺陷。与野生型和互补菌株相比,该突变体对低温的敏感性也有所增加。酵母互补试验表明,VdNop12 能够在 15°C 下功能性恢复酿酒酵母 ΔScNop12 突变体的生长表型。我们证明 VdNop12 定位于细胞核,其缺失导致大丽轮枝菌中几个与 cAMP-PKA 和 MAPK 途径相关的基因表达下调。我们的结果表明 RBPs 在大丽轮枝菌的形态调控、冷适应和致病发育中起关键作用。