菌物学报2026，Vol.45Issue(6)：189-209,21.DOI:10.13346/j.mycosystema.260011
PEG介导的环链虫草菌遗传转化体系建立及hsp30基因抗逆功能分析
Establishment of a PEG-mediated genetic transformation system for Cordyceps cateniannulata and analyses of hsp30 gene function in stress tolerance
摘要
Abstract
Cordyceps cateniannulata is an entomopathogenic fungus with significant biocontrol potential against agricultural and forestry pests.Although the transcriptome sequencing of this fungus has been completed,the lack of establishment of a genetic transformation system has severely hindered functional gene research.In this study,protoplasts of C.cateniannulata were successfully prepared and a PEG-mediated protoplast genetic transformation system was established,achieving stable expression of the exogenous GFP gene.Two novel hsp30 genes,Cchsp30a and Cchsp30b,were cloned from C.cateniannulata(GenBank accession numbers:PX352604 and PX352605,respectively).Using the established transformation system,overexpression mutant strains of Cchsp30a and Cchsp30b were obtained.Phenotypic analysis revealed that,compared to the wild-type strain,the mutant strains exhibited significant enhancement of spore germination rates under heat shock(48 ℃),oxidative stress(menadione),salt stress(NaCl),and cell wall stress(Congo red),and enhanced germ tube elongation under UV radiation.These results confirm the positive regulatory role of Cchsp30 genes in broad-spectrum stress tolerance.Further transcriptome analysis under heat stress showed that a large number of differentially expressed genes(DEGs)in the mutants were significantly enriched in stress-related pathways,particularly the ABC transporter pathway.Notably,genes belonging to the ABCC transporter subfamily were most prominently differentially expressed in the mutants,suggesting that Hsp30 might enhance cellular stress adaptation by modulating the transcription of ABCC subfamily members,thereby influencing substance transport.In summary,this study not only established a genetic manipulation system for C.cateniannulata,but also but also preliminarily characterized the broad-spectrum stress resistance function of Cchsp30 and revealed its potential involvement in stress adaptation through regulating the transcription of ABCC subfamily transporters.These findings provide clues for further exploring the role of Cchsp30 in the stress resistance mechanisms of this entomopathogenic fungus.关键词
PEG 介导/环链虫草/原生质体/遗传转化/hsp30基因/ABC 转运蛋白Key words
PEG-mediated/Cordyceps cateniannulata/protoplast/genetic transformation/hsp30 gene/ATP-binding cassette(ABC)transporters引用本文复制引用
王定锋,李良德,李慧玲,周文鲜,张辉..PEG介导的环链虫草菌遗传转化体系建立及hsp30基因抗逆功能分析[J].菌物学报,2026,45(6):189-209,21.基金项目
福建省自然科学基金(2022J01477) （2022J01477）
福建省农业科学院青年英才项目(YC2021008) This work was supported by the Natural Science Foundation of Fujian Province(2022J01477)and the'Fund for Hundred'Talent Program of Young Science Elite of the Fujian Academy of Agricultural Sciences(YC2021008). （YC2021008）
