浙江大学学报（农业与生命科学版）2026，Vol.52Issue(2)：294-303,10.DOI:10.3785/j.issn.1008-9209.2025.03.212
柑橘间座壳菌绿色荧光蛋白标记菌株的构建及致病性分析
Construction and pathogenicity analysis of green fluorescent protein(GFP)-tagged strains of Diaporthe citri
摘要
Abstract
Diaporthe citri,an important pathogen of citrus,poses a threat to citrus growth and fruit storage.In this study,a genetic transformation system for Diaporthe citri was developed using polyethylene glycol(PEG)-mediated transformation.A filamentous fungal expression vector,pBARGPE1-Hygro,carrying green fluorescent protein(GFP)and hygromycin resistance genes,was successfully introduced into Diaporthe citri strain ZJUQ7.Building on previous fungal transformation techniques,we established an efficient genetic transformation system for Diaporthe citri.Following hygromycin and green fluorescence screening,biological characterization,fluorescence stability verification,and pathogenicity assays of the transformants,34 stable fluorescent strains were obtained.These strains exhibited hyphal morphology,growth rate,and pathogenicity consistent with the wild-type strain.The hyphae of the ZJUQ7-2 transformant were inoculated onto leaves of Newhall Navel orange,and visible lesions appeared after 3 d.These lesions gradually darkened and expanded,eventually leading to the decay of the entire leaf.Fluorescence microscopy of the inoculation site clearly revealed the infection process of Diaporthe citri.This study successfully constructed a protoplast-mediated genetic transformation system for Diaporthe citri and generated stable GFP-labeled strains,providing key tools and technical support for elucidating the pathogen's life cycle,disease progression,and pathogenic mechanism.关键词
柑橘间座壳菌/柑橘黑点病/绿色荧光蛋白/遗传转化Key words
Diaporthe citri/citrus melanose/green fluorescent protein/genetic transformation分类
农业科技引用本文复制引用
牛帆,闫晋珩,程苗嘉,王璟睿,王敏慧,肖小娥,李红叶,焦晨..柑橘间座壳菌绿色荧光蛋白标记菌株的构建及致病性分析[J].浙江大学学报（农业与生命科学版）,2026,52(2):294-303,10.基金项目
国家自然科学基金项目(32470228) （32470228）
浙江省自然科学基金项目(LR25C020001) （LR25C020001）
国家现代农业(柑橘)产业技术体系建设专项(CARS-26). （柑橘）
