草菇Vvrin1基因RNAi转化子的筛选及其转录组分析  

Transformation and transcriptomics analyses of Volvariella volvacea after RNA interference of the Vvrin1 gene

作　　者：陈天赐 陈娟钦 杨国辉 谢宝贵[2] 江玉姬[1,2] 陈炳智[1,2] CHEN Tian-Ci;CHEN Juan-Qin;YANG Gou-Hui;XIE Bao-Gui;JIANG Yu-Ji;CHEN Bing-Zhi(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;Yongtai Agricultural and Rural Bureau,Fuzhou,Fujian 350700,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]永泰县农业农村局,福建福州350700

出　　处：《菌物学报》2021年第12期3265-3275,共11页Mycosystema

基　　金：国家自然科学基金(31801920);福建省自然科学基金(2020J01557);国家现代农业产业技术体系建设专项资金(CARS24)。

摘　　要：草菇味道鲜美,食用价值高,是我国主要栽培的商业食用菌之一。MADS-box转录因子对真核生物的生长发育和信号传导具有关键性的调控作用。本研究通过农杆菌介导的方法获得5个草菇MADS-box转录因子Vvrin1基因的RNA干扰转化子,并进一步对转化子的表型进行分析,发现5个转化子菌丝在PDA固体培养基和栽培料中的生长速度均显著小于(P<0.05)转化野生型菌株H1521,且转化子菌丝颜色呈黄白色,较粗,较稀疏。出菇实验发现,RNA干扰转化子生长停滞在菌丝阶段未能形成原基出菇。转化子菌丝阶段的转录组测序数据发现,上调基因主要富集在核糖体、氨基酸生物合成和次生代谢物的生物合成等途径;下调基因主要富集在MAPK信号通路、脂肪酸氧化、磷脂酰肌醇信号系统等途径。进一步推测MADS-box转录因子Vvrin1基因表达水平降低会影响MAPK信号通路和磷脂酰肌醇信号通路的表达,进而降低菌丝生长速度,影响原基形成。Straw mushroom Volvariella volvacea is delicious and has high edible value.It is one of the main commercial edible fungi cultivated in China.MADS-box transcription factors play a key role in regulating eukaryotic growth and signal transduction.In this study,five RNA interference transformants of the MADS-box transcription factor Vvrin1 gene of V.volvacea were obtained by Agrobacterium-mediated method,and the phenotype of the transformants was further analyzed.It was found that the mycelium growth rate of five transformants was significantly lower(P<0.05)than that of the wild-type strain H1521 on the PDA medium and culture compost,and the mycelia of the transformants were yellowish-white,thicker,and sparse.Fruiting experiment found that the growth of RNA interference transformant was stagnant and did not form primordium.Transcriptomics analysis of the mycelia of transformants showed that the up-regulated genes were mainly enriched in the pathways of ribosome,biosynthesis of amino acids,and secondary metabolites and so on.The down-regulated genes were mainly enriched in MAPK signaling pathway,fatty acid degradation and phosphatidylinositol signaling system.It is inferred that the decreased expression of MADS-box transcription factor Vvrin1 will affect the expression of MAPK signaling pathway and phosphatidylinositol signaling system,and then reduce the mycelial growth rate and affect the primordium formation.

关 键 词：草菇 MADS-BOX 转录因子 RNA 干扰 转录组学 

分 类 号：S646.13[农业科学—蔬菜学]