灵芝漆酶基因家族的克隆及其在不同发育阶段的表达分析  

Cloning and Expression Analysis of Laccase Genes Family at Different Developmental Stages of Ganoderma lucidum

作　　者：刘冬梅 刁文彤 孙雪言 亓希武[1] 梁呈元[1] LIU Dong-mei;DIAO Wen-tong;SUN Xue-yan;QI Xi-wu;LIANG Cheng-yuan(Jiangsu Key Lab.for the Res.&Util.of Plant Res,Inst.of Botany,Jiangsu Prov.&Chinese Acad.of Sci.,Nanjing 210014;Schl.of Environ.Eco.&Bio-Engin,Wuhan Inst.of Technol.,Wuhan 430205)

机构地区：[1]江苏省中国科学院植物研究所江苏省植物资源研究与利用重点实验室,江苏南京210014 [2]武汉工程大学环境生态与生物工程学院,湖北武汉430205

出　　处：《微生物学杂志》2025年第1期34-42,共9页Journal of Microbiology

基　　金：国家自然科学基金青年基金项目(32102457)。

摘　　要：为了明确漆酶基因在灵芝生长发育过程中的作用,克隆了灵芝的15个漆酶基因,并对其进行了生物信息学分析,采用qRT-PCR检测了15个漆酶基因在灵芝不同发育阶段的表达水平。结果表明这些漆酶基因的核苷酸序列长度在1814~2570个碱基对之间,含有4~12个内含子,编码517~628个氨基酸;除Glac14外,其余漆酶基因均含有信号肽;15个漆酶的二级结构均以无规则卷曲和延伸链为主;所有灵芝漆酶均含有Cu-oxidase、Cu-oxidase_2和Cu-oxidase_3保守结构域;Gllac1、Gllac8和Gllac10在菌丝阶段高表达,表明它们在营养生长阶段发挥重要作用;Gllac2、Gllac4、Gllac5、Gllac6、Gllac7、Gllac9、Gllac12和Gllac13在成熟子实体阶段高表达,表明它们可能与子实体的分化和成熟有关;Gllac3、Gllac11、Gllac14和Gllac15在整个发育阶段均有表达,但差异不显著,表明它们可能在灵芝整个发育阶段均发挥作用。本研究为进一步阐明漆酶基因在灵芝生长发育阶段的作用及开发应用漆酶提供参考。In order to clarify the role of laccase genes in the growth and development of Ganoderma lucidum,in this study,15 laccase genes of G.lucidum were cloned and bioinformatically analyzed,and the expression levels of the 15 laccase genes were detected at different developmental stages by qRT-PCR.The results showed that the nucleotide sequences of these laccase genes ranged from 1814-2570 base pairs in length,contained 4-12 introns,and encoded 517-628 amino acids.Except for Glac14,all laccase genes contained signal peptides.The secondary structures of the 15 laccase proteins mainly were irregular coil and extended strand.All laccase proteins contained conservative domains Cu-oxidase,Cu-oxidase_2,and Cu-oxidase_3.Gllac1,Gllac8 and Gllac10 were highly expressed in the mycelium stage,indicating that they play important roles in the nutrient growth stage.Gllac2,Gllac4,Gllac5,Gllac6,Gllac7,Gllac9,Gllac12 and Gllac13 were highly expressed at the stage of mature fructification,suggesting that they may be related to fructification differentiation and maturation.Gllac3,Gllac11,Gllac14 and Gllac15 were expressed throughout the developmental stages,but the differences were not significant,suggesting that they may play roles throughout the developmental stages of G.lucidum.This study lays a foundation for further elucidating the role of laccase genes in the developmental stages of G.lucidum and developing the application of laccase.

关 键 词：灵芝 漆酶 基因克隆 子实体发育 表达水平 

分 类 号：Q933[生物学—微生物学]