漆酶同工酶基因在斑玉蕈生长发育过程中的表达分析  

Expression analysis of laccase isozyme gene during Hypsizygus marmoreus growth and development

作　　者：陈辉[1] 郝海波 赵静 王倩[1] 隽加香 陈明杰[1] 冯志勇[1,3] 张津京[1] CHEN Hui;HAO Hai-Bo;ZHAO Jing;WANG Qian;JUAN Jia-Xiang;CHEN Ming-Jie;FENG Zhi-Yong;ZHANG Jin-Jing(Institute of Edible Fungi,Shanghai Academy of Agricultural Sciences,Shanghai Key Laboratory of Agricultural Genetics and Breeding,Key Laboratory of Resources and Utilization of Edible Fungi(South),Ministry of Agriculture,National Engineering Research Center of Edible Fungi,Shanghai 201403,China;Shanghai Bright Esunyes Bio-tech Co.,Ltd.,Shanghai 201408,China;College of Life Sciences,Nanjing Agricultural University,Nanjing,Jiangsu 210014,China)

机构地区：[1]上海市农业科学院食用菌研究所,上海市农业遗传育种重点开放实验室,农业部南方食用菌资源利用重点实验室,国家食用菌工程技术研究中心,上海201403 [2]上海光明森源生物科技有限公司,上海201408 [3]南京农业大学生命科学学院,江苏南京210014

出　　处：《菌物学报》2020年第6期1038-1048,共11页Mycosystema

基　　金：国家自然科学基金(31601802)。

摘　　要：为了探明漆酶在斑玉蕈生长发育过程中的功能,对斑玉蕈转录测序预测的13个漆酶基因序列进行分析、鉴定和构建分子系统发育树;检测了不同生长发育时期漆酶的活性和漆酶基因表达水平。研究结果显示:13个基因片段中有10个是漆酶基因。不同的漆酶同工酶之间进化关系存在明显差异,大多数漆酶与木腐菌(金针菇Flammulina filiformis和侧耳属Pleurotus)进化关系较近。对斑玉蕈不同生长发育时期的酶活检测结果显示,从斑玉蕈的菌丝恢复期到钉头期,漆酶活性逐渐升高,而在子实体形成后期酶活逐渐降低。对培养40d、60d和80d的菌丝样品以及不同生长发育时期的样品进RT-q PCR检测,结果显示在菌丝营养生长时期,大多数漆酶基因在第40–60天表达量持续增加1–3倍,而在第60–80天时表达量出现降低的情况。而在生殖生长时期,大多数漆酶基因在转色期或者原基期相对表达量达到最大值,并在子实体期出现降低,这与漆酶活性的检测具有一致性。lcc3、lcc7、lcc8和lcc9在斑玉蕈生殖生长过程中相对表达量出现了10–100倍的上调。这说明从菌丝培养到菌丝扭结形成子实体和子实体发育的过程中,不同的漆酶可能发挥着不同的作用,表达量较高的漆酶基因可能对基质降解和子实体形成起主要作用。The function of laccase at different developmental stages of Hypsizygus marmoreus was clarified.Thirteen laccase gene sequences predicted by transcription sequencing of Hypsizygus marmoreus were analyzed and identified,and a phylogenetic tree was constructed.Laccase activity and gene expression levels in different growth and development stages were detected.The results showed that 10 of the 13 gene fragments were laccase genes.The evolutionary relationship between different laccase isoenzymes is obvious difference.Most laccases are closely related to those of wood rot fungi(Flammulina filiformis and Pleurotus spp.).Enzyme activity detection in different growth and development stages of Hypsizygus marmoreus showed that the laccase activity increased gradually from the mycelial regeneration stage to pinhead stage,while decreased gradually in the later stage of fruiting body formation.RT-qPCR of mycelial samples cultured in 40d,60d,and 80d and samples in different growth and development stages was performed.The results revealed that during the period of vegetative growth of mycelium,the expression of most laccase genes continually increased by 1-3 times in 40-60d,and decreased in 60-80d.However,during reproductive period,the relative expression of most laccase genes reached the maximum level at pigmentation stage or primordium stage,but decreased at fruiting stage,consisting with the detection result of laccase activity.The relative expression of lcc3,lcc7,lcc8 and lcc9 increased by 10-100 times during the reproductive period of Hypsizygus marmoreus,indicating that different laccase might play different roles in the growth process from vegetative mycelium,interwoven mycelium to fruiting body formation and development,while highly expressed laccase genes might play a major role in matrix degradation and fruiting body formation.

关 键 词：斑玉蕈 漆酶基因 子实体形成 表达量 

分 类 号：S646[农业科学—蔬菜学]