基因组学与应用生物学
基因組學與應用生物學
기인조학여응용생물학
GENOMICS AND APPLIED BIOLOGY
2013年
4期
420-427
,共8页
李茹%李洋%李林生%廖素环%白凌云%陈保善
李茹%李洋%李林生%廖素環%白凌雲%陳保善
리여%리양%리림생%료소배%백릉운%진보선
板栗疫病菌%ntl基因%低毒病毒%致病力
闆慄疫病菌%ntl基因%低毒病毒%緻病力
판률역병균%ntl기인%저독병독%치병력
Cryphonectria parasitica%ntl%Hypovirus%Virulence
板栗疫病菌(Cryphonectria parasitica)是引起板栗疫病的病原真菌。在实验室前期研究中,获得了一个不支持病毒复制且丧失致病力的板栗疫病菌紫外诱变突变株UV57。与野生型菌株EP155相比,UV57中检测不到蛋白100472的表达。为研究蛋白100472的功能,我们构建了编码100472蛋白的ntl基因的缺失突变体△ntl及其互补转化株△ntl-com。与野生型EP155相比,△ntl菌株表型不变,但对休眠板栗树枝的致病性明显降低,而互补转化株△ntl-com的致病力与野生型没有区别。ntl基因的缺失不影响低毒病毒CHV1-EP713的复制累积量,但导致参与G蛋白信号传导途径的cpga1、cpgb1、cpgc1和ste12基因以及参与MAPK途径的cpmk1转录水平明显下调。本研究结果为阐明病原真菌致病机制提供了新的知识。
闆慄疫病菌(Cryphonectria parasitica)是引起闆慄疫病的病原真菌。在實驗室前期研究中,穫得瞭一箇不支持病毒複製且喪失緻病力的闆慄疫病菌紫外誘變突變株UV57。與野生型菌株EP155相比,UV57中檢測不到蛋白100472的錶達。為研究蛋白100472的功能,我們構建瞭編碼100472蛋白的ntl基因的缺失突變體△ntl及其互補轉化株△ntl-com。與野生型EP155相比,△ntl菌株錶型不變,但對休眠闆慄樹枝的緻病性明顯降低,而互補轉化株△ntl-com的緻病力與野生型沒有區彆。ntl基因的缺失不影響低毒病毒CHV1-EP713的複製纍積量,但導緻參與G蛋白信號傳導途徑的cpga1、cpgb1、cpgc1和ste12基因以及參與MAPK途徑的cpmk1轉錄水平明顯下調。本研究結果為闡明病原真菌緻病機製提供瞭新的知識。
판률역병균(Cryphonectria parasitica)시인기판률역병적병원진균。재실험실전기연구중,획득료일개불지지병독복제차상실치병력적판률역병균자외유변돌변주UV57。여야생형균주EP155상비,UV57중검측불도단백100472적표체。위연구단백100472적공능,아문구건료편마100472단백적ntl기인적결실돌변체△ntl급기호보전화주△ntl-com。여야생형EP155상비,△ntl균주표형불변,단대휴면판률수지적치병성명현강저,이호보전화주△ntl-com적치병력여야생형몰유구별。ntl기인적결실불영향저독병독CHV1-EP713적복제루적량,단도치삼여G단백신호전도도경적cpga1、cpgb1、cpgc1화ste12기인이급삼여MAPK도경적cpmk1전록수평명현하조。본연구결과위천명병원진균치병궤제제공료신적지식。
Cryphonectria parasitica is a causative fungus responsible for the devastating chestnut blight. In our pre-vious study, a UV-induced mutant UV57 which lacks the ability to support the replication of a hypovirus, was ob-tained. Protein 100472 encoded by ntl gene in C. parasitica was not found in UV57 as compared with the wild-type strain EP155. To investigate the function of this protein, ntl deletion mutants were constructed. The ntl null mutants were indistinguishable from the EP155 in phenotype on PDA plate, but exhibited reduced virulence on chestnut stems. Complementation strain of ntl null mutant with a wild-type copy of ntl gene restored the virulence phenotype. Deletion of ntl gene did not change the accumulation of the hypovirus dsRNA. Transcriptional analysis revealed that deletion of ntl reduced the transcript levels of cpga1, cpgb1, cpgc1, ste12, and cpmk1 that encode Gα, Gβ, Gγ, Ste12, and Cpmk1, respectively, key components of heterotrimeric G-protein pathway. These results add new knowledge toward the understanding of the mechanism of virulence regulation in a pathogenic fungus.