State Key Laboratory of Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 2 Yuanmingyuan West Road, Beijing, 100193, China.
Qian Xinan Branch of Guizhou Provincial Tobacco Company, 60 Ruijin Southern Road, Xingyi, 562499, China.
Appl Microbiol Biotechnol. 2024 Jun 28;108(1):398. doi: 10.1007/s00253-024-13238-8.
Grey mould caused by Botrytis cinerea is a devastating disease responsible for large losses to agricultural production, and B. cinerea is a necrotrophic model fungal plant pathogen. Membrane proteins are important targets of fungicides and hotspots in the research and development of fungicide products. Wuyiencin affects the permeability and pathogenicity of B. cinerea, parallel reaction monitoring revealed the association of membrane protein Bcsdr2, and the bacteriostatic mechanism of wuyiencin was elucidated. In the present work, we generated and characterised ΔBcsdr2 deletion and complemented mutant B. cinerea strains. The ΔBcsdr2 deletion mutants exhibited biofilm loss and dissolution, and their functional activity was illustrated by reduced necrotic colonisation on strawberry and grape fruits. Targeted deletion of Bcsdr2 also blocked several phenotypic defects in aspects of mycelial growth, conidiation and virulence. All phenotypic defects were restored by targeted gene complementation. The roles of Bcsdr2 in biofilms and pathogenicity were also supported by quantitative real-time RT-PCR results showing that phosphatidylserine decarboxylase synthesis gene Bcpsd and chitin synthase gene BcCHSV II were downregulated in the early stages of infection for the ΔBcsdr2 strain. The results suggest that Bcsdr2 plays important roles in regulating various cellular processes in B. cinerea. KEY POINTS: • The mechanism of wuyiencin inhibits B. cinerea is closely associated with membrane proteins. • Wuyiencin can downregulate the expression of the membrane protein Bcsdr2 in B. cinerea. • Bcsdr2 is involved in regulating B. cinerea virulence, growth and development.
灰霉病是由灰葡萄孢(Botrytis cinerea)引起的,是导致农业生产大量损失的破坏性疾病,而灰葡萄孢是一种专性坏死型植物病原真菌。膜蛋白是杀菌剂的重要靶标,也是杀菌剂产品研发的热点。武夷菌素影响灰葡萄孢的通透性和致病性,平行反应监测揭示了膜蛋白 Bcsdr2 的关联,阐明了武夷菌素的抑菌机制。在本工作中,我们生成并鉴定了ΔBcsdr2 缺失和互补突变体灰葡萄孢菌株。ΔBcsdr2 缺失突变体表现出生物膜丧失和溶解,其在草莓和葡萄果实上的坏死定殖减少说明了其功能活性降低。Bcsdr2 的靶向缺失也阻断了菌丝生长、产孢和毒力等方面的几个表型缺陷。通过靶向基因互补,所有表型缺陷都得到了恢复。Bcsdr2 在生物膜和致病性中的作用也得到了定量实时 RT-PCR 结果的支持,结果表明,在ΔBcsdr2 菌株感染的早期阶段,磷脂酰丝氨酸脱羧酶合成基因 Bcpsd 和几丁质合酶基因 BcCHSV II 的表达下调。结果表明,Bcsdr2 在灰葡萄孢的各种细胞过程中发挥重要作用。关键点:• 武夷菌素抑制灰葡萄孢的机制与膜蛋白密切相关。• 武夷菌素可以下调灰葡萄孢中膜蛋白 Bcsdr2 的表达。• Bcsdr2 参与调节灰葡萄孢的毒力、生长和发育。