华北农学报2025，Vol.40Issue(5)：188-197,10.DOI:10.7668/hbnxb.20195746
PgβGlu4基因调控大麦条纹病菌致病性的功能分析
Analysis of the Function of PgβGlu4 Gene to Regulate Pathogenicity in Pyrenophora graminea
摘要
Abstract
To investigate the function of the β-glucosidase(βGlu)gene PgβGlu4 from Pyrenophora graminea,which previous studies found to be highly expressed during the infection stage,we constructed a subcellular localiza-tion vector of PCE2-EGFP-PgβGlu4 and transformed rice protoplasts,observed the fluorescence distribution and ana-lyzed its location of existence.Simultaneously,the PgβGlu4 gene RNAi vector was constructed,and QWC protoplasts were prepared by CaCl2-PEG4000 mediated method for genetic transformation.The function of PgβGlu4 gene was studied by detecting the vegetative growth and pathogenicity of the RNAi mutants.Phylogenetic analysis of PgβGlu4 and other homologous proteins from different pathogens showed that PgβGlu4 had a closer evolutionary relationship with that from Pyrenophora tritici-repentis.The subcellular localization results showed that PgβGlu4 was mainly local-ized in the nucleus and cell membrane.Four PgβGlu4 gene RNAi mutants were verified by hygromycin.qRT-PCR a-nalysis showed that the expression of PgβGlu4 gene in four RNAi mutants decreased by 66.31％,68.60％,54.37％and 69.89％,respectively,compared with the wild isolate.The colony diameter was smaller than that of the wild iso-late,and their incidence rate was reduced by 56.69,52.76,47.43,and 53.30 percentage points.After infection with the mutant strain of RNAi-PgβGlu4,the relative chlorophyll content in barley leaves ranged from 30.3 to 35.0,which was significantly higher than that of the wild-type group.The effect of PgβGlu4 gene silencing on the height of barley plants before and after infection was significant compared with that of the wild-type.The results indicated that the PgβGlu4 gene was involved in the regulation of the growth,development,and pathogenicity of Pyronophora graminea.关键词
大麦条纹病菌/β-葡萄糖苷酶/RNA干扰/致病性Key words
Pyrenophora graminea/β-glucosidase/RNA interference(RNAi)/Pathogenicity分类
农业科技引用本文复制引用
杨文娟,祁天涛,王艳婷,孟亚雄,汪军成,王化俊,司二静..PgβGlu4基因调控大麦条纹病菌致病性的功能分析[J].华北农学报,2025,40(5):188-197,10.基金项目
国家自然科学基金项目(32160496 （）
32460517) （）
国家大麦青稞产业技术体系(CARS-05-03B-03) （CARS-05-03B-03）
甘南科技计划项目(22CX8NA047) （22CX8NA047）
甘肃省省级大学生创新创业训练计划项目(S202410733033) （S202410733033）
