粉红螺旋聚孢霉67-1短链脱氢酶基因CrSdr功能研究  

Function of Short-chain Dehydrogenase Gene CrSdr in Clonostachys rosea 67-1

作　　者：陈莹莹 王亚楠 吕斌娜[2] 谭晓东 李世东[2] 孙漫红[1] 马桂珍 CHEN Yingying;WANG Yanan;LüBinna;TAN Xiaodong;LI Shidong;SUN Manhong;MA Guizhen(Jiangsu Key Laboratory of Marine Bioresources and Eco-environment,Jiangsu Ocean University,Lianyungang 222000,China;Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China;Beijing Soil Fertilizer Workstation,Beijing 100029,China)

机构地区：[1]江苏海洋大学江苏省海洋生物资源与生态环境重点实验室,连云港222000 [2]中国农业科学院植物保护研究所,北京100193 [3]北京市土肥工作站,北京100029

出　　处：《中国生物防治学报》2021年第4期750-761,共12页Chinese Journal of Biological Control

基　　金：国家重点研发计划(2019YFD1002000);现代农业产业体系(CARS-23-D05);青海省科技计划(2019-NK-116);中国农业科学院重大产出科研选题(CAAS-ZDXT2018005)。

摘　　要：本研究从实验室前期构建的粉红螺旋聚孢霉67-1寄生核盘菌转录组中获得1个差异表达的短链脱氢酶编码基因CrSdr。实时荧光PCR定量监测显示,菌核诱导下24 h时CrSdr基因的表达水平提高4倍。通过基因敲除与回补研究其功能结果表明,CrSdr敲除后对菌株的生长没有影响,但产孢量比野生菌株提高了43.2%,并且对NaCl、KCl和山梨醇引起的渗透压胁迫更为敏感。敲除突变株对番茄灰霉病菌、大豆菌核病菌和黄瓜枯萎病菌的拮抗作用明显减弱(P<0.05),对核盘菌菌核的寄生能力由野生型菌株的4级降为2级,温室对菌核病的防效降低了50.5%,基因回补后生防作用恢复,表明短链脱氢酶基因在粉红螺旋聚孢霉寄生及生防过程中起着重要的作用。本研究将为揭示粉红螺旋聚孢霉菌寄生机制奠定基础,并对高效植病生防真菌菌剂的研发具有一定的指导意义。A differentially expressed short-chain dehydrogenase gene,CrSdr,was identified from the transcriptome of Clonostachys rosea 67-1 mycoparasitizing the sclerotia of Sclerotinia sclerotiorum,which was previously constructed in our lab.Quantitative real-time PCR detection showed that the expression level of CrSdr increased 4 times after the induction of sclerotia for 24 h.Gene knock-out and complementation were conducted to investigate the function of CrSdr.The results showed that the gene-deleted mutants had the same growth speed with the wild type,spore yields increased by 43.2%,and they were more sensitive to the osmotic pressures of NaCl,KCl and sorbitol.In gene-deleted mutants,the antagonistic activities to Botrytis cinerea,S.sclerotiorum and Fusarium oxysporum f.sp.cucumerinum significantly decreased(P<0.05),and their parasitic ability to S.sclerotiorum sclerotia dropped from 4th of the wild type to 2nd degree.In greenhouse,the control efficiencies against soybean Sclerotinia stem rot decreased by 50.5%compared to the wild type.However,after the gene CrSdr was complemented,the biocontrol activities of 67-1 were recovered,suggesting that CrSdr plays important roles in mycoparasitism and biological control of C.rosea.The research provides a foundation for understanding the mechanism of C.rosea mycoparasitism,and will be helpful for the development of highly efficient biocontrol fungal agents against plant diseases.

关 键 词：粉红螺旋聚孢霉 短链脱氢酶 菌寄生作用 核盘菌 基因敲除与回补 荧光定量PCR 

分 类 号：S476[农业科学—农业昆虫与害虫防治]