RNAi法分析香菇YUCCA8基因的功能  

Functional Analysis of Indole-3-pyruvate Monooxygenase Gene YUCCA8 Involved in Heat Resistance of Lentinula edodes by RNAi

作　　者：钟海英 罗义 龚钰华[1] 徐瑞平 边银丙[1] ZHONG Haiying;LUO Yi;GONG Yuhua;XU Ruiping;BIAN Yingbing(Institute of Applied Fungi,Huazhong Agricultural University,Wuhan 430070,Hubei,China)

机构地区：[1]华中农业大学植物科学技术学院应用真菌研究所,湖北武汉430070

出　　处：《食用菌学报》2021年第3期11-18,共8页Acta Edulis Fungi

基　　金：国家自然科学基金(31972476、32072641)。

摘　　要：构建香菇(Lentinula edodes)YUCCA 8基因的双向启动子(Leactin和Legpd)RNA干扰载体,通过根癌农杆菌介导的香菇菌丝转化方法,获得12个相对稳定的YUCCA 8基因沉默阳性转化子。qRT-PCR分析表明,转化子的YUCCA 8基因表达量下调至野生型菌株S606的10%~40%;与S606相比,转化子菌丝生长显著加快,菌丝耐热性变弱,菌丝IAA含量降低。研究表明,香菇YUCCA 8基因可控制菌丝IAA的合成且可影响香菇菌丝耐热性。In plants,3-indolepyruvate monooxygenase(YUCCA8)is a rate-limiting enzyme for the biosynthesis of the major auxin indole-3-acetic acid(IAA).Previous studies found that endogenous IAA level affected heat resistance in Lentinula edodes.In this paper,the RNA interference technology was used to analyze the function of YUCCA8 in L.edodes.The thermotolerant L.edodes strain S606 was used to be silenced for YUCCA 8 by a two-way promoter(Leactin and Legpd)RNAi vector.Through Agrobacterium tumefaciens mediated transformation of L.edodes hyphae,12 relatively stable YUCCA8-silenced positive transformants were obtained.The results of qRT-PCR showed that expression levels of YUCCA8 in the 12 transformants were 10%-40%of that in S606.Compared with S606,the transformants showed significantly faster mycelial growth,weaker tolerance to heat stress and a lower IAA content.The results indicate that YUCCA 8 controls the biosynthesis of IAA in L.edodes hypha and facilitates heat resistance in L.edodes.

关 键 词：香菇 耐热性 吲哚-3-乙酸 YUCCA8 

分 类 号：S646.12[农业科学—蔬菜学]