菌物学报2026，Vol.45Issue(8)：94-103,10.DOI:10.13346/j.mycosystema.260003
五味子叶枯病菌细极链格孢GFP荧光标记及侵染过程
Green fluorescent labeling and infection process of Alternaria tenuissima causing leaf blight in Schisandra chinensis
摘要
Abstract
Leaf blight of Schisandra chinensis,caused by Alternaria tenuissima,is an important disease in commercial fields.To observe the infection process of the pathogen,a GFP fluorescent labeling vector pBS-HYG-GFP was constructed using homologous recombination technology and the protoplast preparation conditions were optimized(hyphae aged 36 h enzymolysed with 0.01 g/mL driselase,0.01 g/mL lyticase and 0.01 g/mL snailase at 28℃for 2.5 h).The protoplast yield reached 1.14×106/mL.Following genetic transformation,a stable fluorescent strain SccA071GFP was obtained that exhibited no significant differences from the wild-type strain in terms of growth rate,spore production capacity and pathogenicity.Infection process observation revealed that conidia attached and germinated on the leaf surface,with germ tubes developing into primary hyphae.The hyphae preferentially extended along the depressions on the leaf surface and mainly invaded the mesophyll tissue through stomata or intercellular spaces in 36 hours after inoculation.In 96 hours after inoculation,the hyphae spread in a network within the mesophyll tissue,and extensively branched and densely colonized the tissue,completing the entire process of penetration,incubation,and symptom development in 120 hours after inoculation.In this study,an efficient and stable genetic transformation system was first established for the pathogen causing leaf blight in S.chinensis.The findings provide a stable and efficient genetic transformation system for identifying functional genes in the pathogen,and advantage in-depth investigations into pathogen infection and pathogenesis mechanisms.关键词
细极链格孢/绿色荧光蛋白标记/遗传转化/侵染过程Key words
Alternaria tenuissima/green fluorescent labeling/genetic transformation/infection process引用本文复制引用
孔德宇,吴丽婷,孙春蕾,李自博,朴静子,周如军..五味子叶枯病菌细极链格孢GFP荧光标记及侵染过程[J].菌物学报,2026,45(8):94-103,10.基金项目
国家重点研发计划(2024YFD1600600)This work was supported by the National Key Research and Development Program of China(2024YFD1600600). （2024YFD1600600）
