双孢蘑菇萎锈灵抗性基因定点突变的载体构建与遗传转化  

Construction of a vector containing a point-mutated carboxin-resistance gene and genetic transformation of Agaricus bisporus

作　　者：卢园萍 肖婷婷 尚俊军 曾志恒[1] 陈美元[1] 鲍大鹏[2] LU Yuan-Ping;XIAO Ting-Ting;SHANG Jun-Jun;ZENG Zhi-Heng;CHEN Mei-Yuan;BAO Da-Peng(Institute of Edible Fungi,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350012,China;Institute of Edible Fungi,Shanghai Academy of Agricultural Sciences,Shanghai 201403,China)

机构地区：[1]福建省农业科学院食用菌研究所,福建福州350012 [2]上海市农业科学院食用菌研究所,上海201403

出　　处：《菌物学报》2021年第12期3256-3264,共9页Mycosystema

基　　金：福建省自然科学基金(2019J05143);福建省农业科学院科研项目(AGP2018-10);国家现代农业产业技术体系专项资金(CARS-20);“5511”协同创新工程(XTCXGC2021007)。

摘　　要：为建立更为安全、有效的双孢蘑菇遗传转化体系,构建了双孢蘑菇琥珀酸脱氢酶的铁硫蛋白亚基Agsdi1突变(His突变为Leu)表达载体pAgsdi1,并通过农杆菌介导方法转化双孢蘑菇W192,经萎锈灵筛选以及PCR扩增和MnlⅠ酶切验证后获得了转化菌株。验证结果表明,点突变的铁硫蛋白亚基Agsdi1可以作为双孢蘑菇有效的抗性标记基因。因其并未引入新的外源基因,是一种比潮霉素抗性基因更为安全的筛选标记,将可用于双孢蘑菇等食用菌的遗传转化。In order to establish a safe and efficient genetic transformation system for Agaricus bisporus,an expressed vector with mutant Agsdi1(a single amino-acid substitution,His to Leu)encoding iron-sulphur protein of succinate dehydrogenase was constructed.After transformation of A.bisporus strain W192 by Agrobacterium-mediated method,putative transformants were selected by carboxin and identified by PCR amplification and digestion with MnlⅠrestriction enzyme.The result showed that a point-mutated Agsdi1 can serve as an effective resistance marker for A.bisporus.Without introducing exogenous gene,point-mutated Agsdi1 gene is a safer selective marker than hygromycin resistance gene,and can be used for genetic transformation of edible mushroom such as A.bisporus.

关 键 词：食用菌 遗传转化 抗性标记 萎锈灵 

分 类 号：S646[农业科学—蔬菜学]