草菇MADS-box转录因子Vvrin1基因过表达转化子菌株的构建及其生长速度的初步差异分析  

Construction of the overexpression transformant strains with MADS-box transcription factor Vvrin1 gene and analysis on differential growth rate of Volvariella volvacea

作　　者：陈炳智[1] 李凌 陈天赐 邱铭锰 吴敏文 谢宝贵[1,2] 江玉姬[1,2] CHEN Bing-Zhi;LI Ling;CHEN Tian-Ci;QIU Ming-Meng;WU Min-Wen;XIE Bao-Gui;JIANG Yu-Ji(College of Food Science,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;Mycological Research Center,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;Fuzhou Agricultural Bureau,Fuzhou,Fujian 350000,China)

机构地区：[1]福建农林大学食品科学学院,福建福州350002 [2]福建农林大学菌物研究中心,福建福州350002 [3]福建省福州市农业局,福建福州350000

出　　处：《菌物学报》2020年第6期1049-1055,共7页Mycosystema

基　　金：国家自然科学基金(31801920);国家现代农业产业技术体系建设专项资金(CARS24);国家重点基础研究发展计划(2014CB138302);福建省教育厅中青年教师科研项目(JT180115);福建农林大学科技创新基金项目(CXZX2017410)。

摘　　要：新鲜草菇味道鲜美,香味浓郁,而且营养丰富,是具有中国特色的食药用菌。草菇采后极易开伞,低温贮藏(10℃以下)则容易自溶、渗水腐、发出异味,是最不易贮藏的食用菌之一。本课题组先前的研究表明草菇MADS-box转录因子Vvrin1基因可能在草菇菌柄的伸长、菌盖的开伞过程中起到一定的作用。因此,本研究以Vvrin1基因为研究对象,构建该基因的过表达载体,通过农杆菌介导的方法进行转化草菇异核菌株H1521菌丝块。通过潮霉素抗性平板筛选,PCR扩增潮霉素抗性基因及Vvrin1基因过表达特异片段,q RT-PCR分析拟转化子菌株内的Vvrin1基因表达量,最终获得了8个比较可靠的转化子。进一步对这些转化子的表型进行初探,发现其中7个转化子的生长速度比出发菌株H1521快,具有显著性差异(P<0.05),且转化子菌丝更浓密,菌落表面颜色更深,推测草菇MADS-box转录因子Vvrin1基因可能参与了菌丝阶段生长速度和色素合成或积累的调控。研究结果为进一步研究草菇MADS-box转录因子Vvrin1基因的功能提供了菌株材料及数据支持。The pileus of Volvariella volvacea easily opens after harvest and easily self-dissolves, oozily rots and smells in cold storage(below 10℃), making the fungus difficult to store. Previous studies showed that the Vvrin1 gene of MADS-box transcription factor might play a certain role in the elongation of stalk and the pileus expansion. In this study, the overexpression vector of Vvrin1 gene was constructed, and the heterokaryotic strain H1521 mycelium block was transformed by agrobacterium-mediated method. Through hyodamycin resistance plate screening, the hyodamycin resistance gene and Vvrin1 gene overexpression special fragment were amplified by PCR, and the Vvrin1 gene expression level in the transformants were analyzed by q RT-PCR, and finally 8 relatively reliable transformants were obtained. Preliminary study of the phenotype of the transformants indicated that in which seven transformants grew faster than the wild strain H1521, showing significant difference(P<0.05), and the hyphae of transformants were denser, with deeper color on colony surface. It could be inferred that MADS-box transcription factor Vvrin1 gene might participate in regulation of hypha growth and pigment synthesis and accumulation in V. volvacea. The results provide the strain materials and data support for further study on the regulation mechanism of mature senescence(especially pileus expansion) of V. volvacea.

关 键 词：草菇 MADS-box转录因子 过表达 生长速度 

分 类 号：S646.13[农业科学—蔬菜学]