根癌农杆菌介导的少孢节丛孢菌遗传转化体系研究  

Study on Agrobacterium tumefaciens-Mediated Transformation System of Arthrobotrys oligospora

作　　者：黄运福 贡莎莎 孟庆玲[1] 乔军[1] 张国武 王熙凤 才学鹏[2] HUANG Yun-fu;GONG Sha-sha;MENG Qing-ling;QIAO Jun;ZHANG Guo-wu;WANG Xi-feng;CAI Xue-peng(Colege of Animal Science and Technology,Shihezi University,Shihezi,Xinjiang,832003,China;China Institute of Veterinary Drug Control,Beijing,100081,China)

机构地区：[1]石河子大学动物科技学院,新疆石河子832003 [2]中国兽医药品监察所,北京100081

出　　处：《动物医学进展》2021年第2期60-65,共6页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31460654,31260601);国家重点研发计划项目(2017YFD0501200)。

摘　　要：根癌农杆菌介导的遗传转化法(ATMT)是一种丝状真菌常用的遗传转化手段,但目前还不能有效地应用于少孢节丛孢菌的转化。构建由PgpdA启动子调控mgfp-gus基因的真菌表达载体pCAMBIA1304'-PgpdA并转入农杆菌中,利用ATMT技术对少孢节丛孢菌进行转化。结果显示,农杆菌AGL-1介导的转化在共培养时间为60 h,温度为23℃的条件下获得了1株转化子,以其基因组为模板可以扩增出hph和gus基因片段,进行GUS组织染色能观察到蓝色,表明目的基因成功转入少孢节丛孢菌基因组中并能正常表达。初步建立了农杆菌介导的少孢节丛孢菌遗传转化方法,为少孢节丛孢菌功能新基因的研究、菌株改良等研究奠定了基础。Agrobacterium tumefaciens-mediated transformation(ATMT)is a common genetic transformation method of filamentous fungi,but it can not be effectively applied to the transformation of Arthrobotrys oligospora at present.In this study,a fungal expression vector pCAMBIA1304'-PgpdA,which regulates the mgfp-gus gene by the PgpdA promoter,was constructed and transferred into Agrobacterium tumefaciens,and then Arthrobotrys oligospora was transformed by ATMT.The result showed that a transformant was obtained by AGL-1 strain-mediated transformation under the co-cultivation condition of 23℃for 60 hours.The hph and gus genes were amplified in the genome,and the blue color was observed by GUS tissue staining,indicating that the target gene had been transferred into the genome of Arthrobotrys oligospora and could be expressed normally.Agrobacterium-mediated transformation of Arthrobotrys oligospora was established,which laid a foundation for the study of new functional genes and strain improvement of Arthrobotrys oligospora.

关 键 词：少孢节丛孢菌 根癌农杆菌 遗传转化 mgfp-gus基因 

分 类 号：S852.66[农业科学—基础兽医学] Q789[农业科学—兽医学]