菌物学报2025，Vol.44Issue(4)：59-67,9.DOI:10.13346/j.mycosystema.240283
越南假拟盘多毛孢转化体系构建
Construction of a transformation system for Pseudopestalotiopsis vietnamensis
摘要
Abstract
Pseudopestalotiopsis vietnamensis is widespread in the tea gardens in Wuyi Mountain,causing gray blight disease(GBD)that severely affects the quality of tea.For understanding the pathogenic mechanism of Ps.vietnamensis,the research on protoplast transformation system of the fungus was conducted.Protoplasts of purified Ps.vietnamensis were isolated by using an enzymatic digestion method.pSULPH plasmid was employed as a vector,and the target plasmid was introduced into the protoplasts via PEG-Ca2+-mediated transformation to promote its regeneration.Finally,the GFP-transformed strains were validated through confocal laser scanning microscopy and protein extraction.It was found that the mycelia of Ps.vietnamensis exhibited severe clumping on CM medium.The clumping appearance could be alleviated by switching the mycelia to PD medium,and thereby improving the efficiency of enzymatic digestion and providing strong support for obtaining high-quality protoplasts.Using the PEG-Ca2+-mediated transformation method,the green fluorescent protein gene was successfully introduced into Ps.vietnamensis strain CLBB1,resulting in the acquirement of transformant with uniform fluorescence intensity and genetic stability.This study lays the foundation for further research into the functions of Ps.vietnamensis pathogenic genes.关键词
越南假拟盘多毛孢/原生质体/PEG-Ca2+介导/GFP转化/茶轮斑病Key words
Pseudopestalotiopsis vietnamensis/protoplast/PEG-Ca2+-mediated/GFP transformation/gray blight disease引用本文复制引用
刘午鸿,平均雄,齐赛洋,罗心阳,郭依娜,戴方妍,傅仙玉,武广珩..越南假拟盘多毛孢转化体系构建[J].菌物学报,2025,44(4):59-67,9.基金项目
南平市资源化学产业科技创新联合资助项目(N2020Z007) （N2020Z007）
福建省自然科学基金(2022J011200)This work was supported by the Joint Funding Project for Science and Technology Innovation of Nanping Resource Chemical Industry(N2020Z007)and the Natural Science Foundation of Fujian Province(2022J011200). （2022J011200）
