转录组测序筛选克氏原螯虾卵巢发育、免疫和生长相关基因  

Transcriptome analysis of Procambarus clarkii to screen genes related to ovary development, immunity and growth

作　　者：江红霞 刘慧芬 马晓 吴利敏 孔祥会 李学军 JIANG Hongxia;LIU Huifen;MA Xiao;WU Limin;KONG Xianghui;LI Xuejun(Engineering Lab of Henan Province for Aquatic Animal Disease Control,Engineering Technology Research Center of Henan Province for Aquatic Animal Cultivation,College of Fisheries,Henan Normal University,Xinxiang 453007,China)

机构地区：[1]河南师范大学,水产学院水产动物疾病控制河南省工程实验室,河南省水产动物养殖工程技术研究中心,河南新乡453007

出　　处：《水产学报》2021年第3期396-414,共19页Journal of Fisheries of China

基　　金：国家自然科学基金(31972812);河南省重点科技攻关项目(192102110081);河南师范大学博士科研启动费支持项目(qd17142);河南师范大学个人科研项目结余经费资助专项(20180531)

摘　　要：为发掘克氏原螯虾卵巢发育、免疫和肌肉生长的重要功能基因,采用Illumina HiSeqTM 2500高通量测序平台对克氏原螯虾的卵巢、肝胰腺和肌肉组织进行了转录组测序。所得序列经质控、组装后比对到NR、Swiss-Prot、pfam、COG、GO和KEGG数据库中注释,并进行差异基因聚类分析。结果显示,测序共获得了53006个unigene,平均长度为1194 bp。对3个组织样品的测序文库进行两两比较,发现在卵巢vs.肝胰腺中有差异表达基因(differentially expressed gene,DEG)20382个,在肝胰腺vs.肌肉中有DEG 12753个,在肌肉vs.卵巢中有DEG 21629个。GO功能分类分析发现,部分DEG被注释到繁殖(reproduction)、繁殖过程(reproduction process)、免疫系统过程(immune system process)和生长(growth)GO条目。KEGG pathway分析显示,一部分DEG在卵巢发育、免疫和肌肉生长相关的信号通路中得到了富集。根据GO功能分类和KEGG信号通路分析筛选出了大量与克氏原螯虾卵巢发育、免疫和肌肉生长相关的候选基因,如卵黄蛋白原、卵黄蛋白原受体、Toll样受体2、Toll样受体相互作用蛋白、肌肉生长抑制素和5-羟色胺受体等。本研究结果丰富了克氏原螯虾的基因资源,可为克氏原螯虾的遗传育种和免疫研究提供基础数据。Procambarus clarkii is one of the freshwater lobsters which has important economic value in China.However,long-term high-density farming and inbreeding have brought many negative effects on the cultivation of P.clarkii,such as slow growth,small size,frequent diseases,and the difficulty of artificial breeding caused by the asynchrony of gonadal development,which have severely restricted the further development of P.clarkii aquaculture.In order to find candidate genes involved in ovary development,immunity and muscle growth of P.clarkii for improving the production performance of this species,the transcriptomes of ovary,hepatopancreas and muscle of P.clarkii were sequenced by a new generation of high throughput sequencing technology.After quality control and assembly,the sequences acquired were blasted against NR,Swiss-Prot,pfam,COG,GO and KEGG databases,and then cluster analyses were performed.In total,53006 final unigenes with an average length of 1194 bp were obtained.Pairwise comparison of sequencing libraries of 3 tissue samples revealed that 20382 differentially expressed genes(DEGs)were found in ovary vs.hepatopancreas group,12753 DEGs were found in hepatopancreas vs.muscle group,21629 DEGs were found in muscle vs.ovary group.Gene ontology analysis indicated that some DEGs were annotated into"reproduction","reproduction process","immune system process"and"growth"GO terms.KEGG pathway analysis showed that some DEGs were enriched in signaling pathways related to ovarian development,immunity and muscle growth.Based on GO functional classification and KEGG pathway analysis,a large number of candidate genes related to ovarian development,immunity and muscle growth of P.clarkii were screened,such as vitellogenin,vitellogenin receptor,Toll-like receptor 2,Toll-interacting protein,myostatin,5-hydroxytryptamine receptor,etc.The results of this study enriched the gene resources of P.clarkii and could provide basic data for the further genetic breeding and immunity research of this commercial species.

关 键 词：克氏原螯虾 转录组测序 差异表达基因 功能注释 

分 类 号：S917.4[农业科学—水产科学]