雪峰虫草子座不同生长发育时期的转录组研究  

Transcriptomic Analysis of Ophiocordyceps xuefengensis Stromata at Different Development Stages

作　　者：钟灿[1,2] 金剑 刘浩 谢景[1] 刘平安 张水寒 ZHONG Can;JIN Jian;LIU Hao;XIE Jing;LIU Ping-an;ZHANG Shui-han(Institute of Chinese Materia Medica,Hunan Academy of Chinese Medicine,Changsha 410013,China;State Key Laboratory of Dao-di Herbs,Beijng 100700,China)

机构地区：[1]湖南省中医药研究院中药研究所,湖南长沙410013 [2]道地药材国家重点实验室,北京100700

出　　处：《中国现代中药》2023年第3期523-532,共10页Modern Chinese Medicine

基　　金：国家自然科学基金项目(81673585);湖南省自然科学基金项目(2020JJ5330);2017年中医药公共卫生服务补助专项(财社〔2017〕66号);中央本级重大增减支项目(2060302);湖南省高层次卫生人才“225”工程培养项目(湘卫函〔2019〕196号);湖南省科技重大专项(2014FJ1007)。

摘　　要：目的:挖掘调控雪峰虫草生长发育的相关通路和基因,揭示其子座生长发育的内在分子机制。方法:采用转录组测序技术(RNA-Seq)对雪峰虫草子座菌丝体(MD1)、子座原基(PD2)和子座(SD3)生长发育3个不同阶段进行测序;数据通过非冗余蛋白(NR)、Swiss-Prot、Pfam、京都基因与基因组百科全书(KEGG)、真核生物相邻类的聚簇(KOG)、基因本体(GO)数据库进行功能注释。利用EdgeR软件进行差异表达基因分析,挖掘调控雪峰虫草子座生长发育的相关通路和基因。结果:测序结果显示,MD1、PD2、SD3分别获得了8.15、7.51、6.87GB分析数据,拼接分析数据共获得34218个转录本,平均长度为5502bp,组装及去冗余后获得10511条unigenes,平均长度为2470bp。KOG功能注释和GO富集分析显示,细胞和细胞组分、新陈代谢过程、信号转导及碳代谢是雪峰虫草子座发育过程中的关键基因类别。对子座不同生长发育时期的基因进行差异表达分析发现,PD2和SD3相比差异表达基因最多,其中上调基因1073个,下调基因1036个,MD1和SD3相比差异表达基因最少,上调基因533个,下调基因578个。差异表达基因的KEGG通路富集结果显示,细胞外信号调节激酶(MAPK)信号通路是参与调节雪峰虫草子座生长发育的关键信号通路。结论:Ste2和Gpd1及其所在途径可能分别调控雪峰虫草子座原基的形成和子座的生长。Objective:To study the molecular mechanism of the growth and development of Ophiocordyceps xuefengensis stromata.Methods:RNA-Seq was employed to determine the sequences and to mine the differentially expressed genes(DEGs)of the mycelium(MD1),primordium(PD2),and stroma(SD3)samples of O.xuefengensis.The obtained data were used for annotation against the Non-Redundant Proteins(NR),Swiss-Prot,Pfam,Kyoto Encyclopedia of Genes and Genomes(KEGG),clusters of EuKaryotic Orthologous Groups(KOG),and Gene Ontology(GO)databases.The differential expressed genes were analyzed by EdgeR software,and the related pathways and genes regulating the growth and development of O.xuefengensis stromata were excavated.Results:From MD1,PD2,and SD3 samples respectively,8.15,7.51,and 6.87 GB clean reads were obtained.A total of 34218 transcripts with an average length of 5502 bp were acquired.After assembly and redundancy removal,10511 unigenes were obtained with an average length of 2470 bp.The KOG annotation and GO enrichment analysis showed that the key genes in the development of O.xuefengensis were involved in cells and cellular parts,metabolic processes,signal transduction,and carbon metabolism.There were 1073 significantly upregulated genes and 1036 significantly down-regulated genes in SD3 as compared with PD2,as well as 533 significantly upregulated genes and 578 significantly down-regulated genes in SD3 as compared with MD1.The KEGG pathway enrichment showed that the DEGs were mainly involved in the MAPK signaling pathway in the growth and development of O.xuefengensis stromata.Conclusion:Ste2 and Gpd1 respectively regulate the formation of the primordium and stroma of O.xuefengensis.

关 键 词：雪峰虫草 人工培育 丝状真菌 

分 类 号：R282.12[医药卫生—中药学]