利用转录组挖掘羊肚菌继代菌丝退化相关基因的初步探讨  

Preliminary Transcriptome Mining of Genes Related to Mycelium Degeneration of Morchella esculenta During Subculture

作　　者：徐爱国 储婷 杨瑞恒[2] 李福后 张建 唐利华[2] XU Aiguo;CHU Ting;YANG Ruiheng;LI Fuhou;ZHANG Jian;TANG Lihua(Institute of Plateau Biology,Tibet Autonomous Region,Lhasa 850000,Tibet,China;Institute of Edible Fungi,Shanghai Academy of Agricultural Sciences,Shanghai 201403,China;School of Food Science and Engineering,Jiangsu Ocean University,Lianyungang 222005,Jiangsu,China)

机构地区：[1]西藏自治区高原生物研究所,西藏拉萨850000 [2]上海市农业科学院食用菌研究所,上海201403 [3]江苏海洋大学食品科学与工程学院,江苏连云港222005

出　　处：《食用菌学报》2023年第1期10-16,共7页Acta Edulis Fungi

基　　金：西藏自治区重点研发计划(XZ202101ZY0005N)。

摘　　要：继代培养7代羊肚菌(Morchella esculenta)菌丝体,计算不同代数的菌丝生长速度,再分别收集第一代(M 1)和第六代(M 6)菌丝体,通过转录组测序,GO功能注释和富集与KEGG通路富集,得到关键差异基因,最后选取7个与退化相关的关键差异基因cel1、lac2、cah、png1、zpr1、did2和Hsp31,采用荧光定量PCR测定基因相对表达量,以验证转录组分析结果的可靠性。结果表明:当继代培养至第7代时,羊肚菌菌丝不能生长;M6与M1相比,共得到575个关键差异基因,其中表达上调198个,表达下调377个;GO富集表明,关键差异基因主要富集在活性氮代谢过程、硝酸盐代谢过程、硝酸盐同化和从头合成次黄嘌呤核苷酸的生物过程;KEGG通路富集表明,关键差异基因主要富集在磷酸戊糖途径、泛酸和乙酰辅酶A生物合成、硒化合物代谢、生物素代谢和精氨酸生物合成通路;7个关键差异基因的相对表达量与转录组测序分析结果一致。The underlying mechanism of mycelium degeneration of Morchella esculenta in the process of subculture was studied through comparative transcriptome analysis.Mycelia of M.esculenta Liumei No.8 were successively subcultured for seven passages,and then mycelial growth rate was calculated for different passages.The mycelia of the first(M1)and the sixth(M6)passage were collected,sequenced and then compared for transcriptome data through GO functional annotation and KEGG pathway enrichment analysis.Eventually,seven differentially expressed genes(DEGs)related to mycelium degeneration were selected,and they were cel1,lac2,cah,png1,zpr1,did2 and Hsp31.These genes were then verified by qRT-PCR.The results showed that mycelia of M.esculenta Liumei No.8 failed to proliferate at the seventh passage.There were 575 DEGs between M1 and M 6,among which 198 DEGs were up-regulated and 377 DEGs were down-regulated.GO enrichment analysis showed that the DEGs were enriched in reactive nitrogen species metabolic process,nitrate metabolic process,nitrate assimilation,and‘De novo’IMP biosynthetic process.KEGG pathway enrichment analysis showed that the DEGs were enriched in pentose phosphate pathway,pantothenate and CoA biosynthesis,selenocompound metabolism,biosynthesis of biotin cofactors,biotin metabolism,and arginine biosynthesis.The relative expression levels of the seven selected DEGs were consistent with the transcriptome analysis.

关 键 词：羊肚菌 菌丝 继代 退化 转录组 

分 类 号：S646.7[农业科学—蔬菜学]