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Abstract：
Coniella vitis is a dominant phytopathogen of grape white rot in China, significantly impacting grape yield and quality. Previous studies showed that the growth and pathogenicity of C. vitis were affected by the environmental pH. Arrestin-like protein PalF plays a key role in mediating the activation of an intracellular-signaling cascade in response to alkaline ambient. However, it remains unclear whether palF affects the growth, development, and virulence of C. vitis during the sensing of environmental pH changes. In this study, we identified a homologous gene of PalF/Rim8 in C. vitis and constructed CvpalF-silenced strains via RNA interference. CvpalF-silenced strains exhibited impaired fungal growth at neutral/alkaline pH, accompanied by reduced pathogenicity compared to the wild-type (WT) and empty vector control (CK) strains. The distance between the hyphal branches was significantly increased in the CvpalF-silenced strains. Additionally, CvpalF-silenced strains showed increased sensitivity to NaCl, H2O2, and Congo red, and decreased sensitive to CaSO4. RT-qPCR analysis demonstrated that the expression level of genes related to pectinase and cellulase were significantly down-regulated in CvpalF-silenced strains compared to WT and CK strains. Moreover, the expression of PacC, PalA/B/C/F/H/I was directly or indirectly affected by silencing CvpalF. Additionally, the expression of genes related to plant cell wall-degrading enzymes, which are key virulence factors for plant pathogenic fungi, was regulated by CvpalF. Our results indicate the important roles of CvpalF in growth, osmotolerance, and pathogenicity in C. vitis.
摘要：
短尾球藻是我国葡萄白腐病的主要植物病原菌,显著影响葡萄产量和品质。先前的研究表明，C.vitis的生长和致病性受到环境pH值的影响。抑制蛋白样蛋白PalF在介导响应碱性环境的细胞内信号传导级联的激活中起关键作用。然而,目前尚不清楚palF是否会影响生长，发展,在感知环境pH变化过程中，C.vitis的毒力。在这项研究中,我们在C.vitis中鉴定了PalF/Rim8的同源基因，并通过RNA干扰构建了CvpalF沉默菌株。CvpalF沉默的菌株在中性/碱性pH下表现出受损的真菌生长，与野生型（WT）和空载体对照（CK）菌株相比，致病性降低。在CvpalF沉默的菌株中，菌丝分支之间的距离显着增加。此外,CvpalF沉默的菌株对NaCl的敏感性增加，H2O2和刚果红,并降低对CaSO4的敏感性。RT-qPCR分析表明，与野生型和CK菌株相比，在CvpalF沉默的菌株中果胶酶和纤维素酶相关基因的表达水平显着下调。此外，PacC的表达式，沉默CvpalF直接或间接影响PalA/B/C/F/H/I。此外,与植物细胞壁降解酶相关的基因的表达，是植物病原真菌的关键毒力因子，受CvpalF调控。我们的结果表明CvpalF在生长中的重要作用，osmotolerance,和C.葡萄的致病性。
