巨须裂腹鱼水霉菌的分离鉴定及其对脾脏转录组的影响  

Isolation,identification and the effect of Saprolegniasis on the spleen transcriptome of Schizothorax macropogon

作　　者：孔庆辉 刘海平 刘锁珠 索朗斯珠[1] 徐业芬 赵晓玲 商振达 KONG Qinghui;LIU Haiping;LIU Suozhu;SUO Langsizhu;XU Yefen;ZHAO Xiaoling;SHANG Zhenda(College of Animal Science,Tibet Agricultural&Animal Husbandry University,Nyingchi 860000,China;Institute of Fisheries Science,Tibet Academy of Agricultural and Animal Husbandry Sciences,Lhasa 850002,China;Tibetan Plateau Feed Processing Research Center,Tibet Agricultural&Animal Husbandry University,Nyingchi 860000,China)

机构地区：[1]西藏农牧学院动物科学学院,西藏自治区林芝860000 [2]西藏自治区农牧科学院水产科学研究所,西藏自治区拉萨850002 [3]西藏农牧学院,西藏高原饲料加工工程研究中心,西藏自治区林芝860000

出　　处：《水产学报》2023年第8期133-142,共10页Journal of Fisheries of China

基　　金：西藏自治区重点研发及转化计划项目(XZ202001ZY0016N);西藏高原饲料加工工程研究中心项目(XZJYT2018GCZX)。

摘　　要：为分析感染水霉菌对巨须裂腹鱼脾脏转录组的影响,探索水霉菌感染巨须裂腹鱼的分子机制,实验随机选取生活在同一水域中的5尾健康和5尾感染水霉菌的巨须裂腹鱼,采用PDA琼脂培养基和分子生物学方法对巨须裂腹鱼水霉菌进行分离鉴定,并采用Illumina HiSeqTM 2000高通量测序平台,对健康和感染水霉菌的巨须裂腹鱼脾脏组织进行转录组测序,并对测序数据进行拼接、注释和差异表达基因分析。结果显示,从巨须裂腹鱼皮肤上分离鉴定得到3株水霉菌;转录组数据显示,健康组和感染水霉病组分别获得46619504条和43912876条数据,与健康组相比,感染水霉菌组共有1889个基因发生差异表达,其中1414个基因上调,475个基因下调,随机选取6个差异表达基因进行实时荧光定量PCR(RT-qPCR)验证,验证结果与转录组测序一致。对健康组和感染水霉菌组的差异表达基因进行GO功能富集发现,上调基因主要富集于241个功能中,下调基因主要富集于60个功能中,上述基因主要涉及分子功能类、细胞组分类和生物过程类等生理功能;KEGG通路富集分析发现,差异表达基因主要富集在免疫疾病、内分泌和代谢疾病、病毒感染性疾病、消化系统及排泄系统等。研究表明,感染水霉菌会影响巨须裂腹鱼脾脏组织多种基因的表达量,实验结果为进一步探索巨须裂腹鱼水霉菌的感染机制奠定了基础。Schizothorax macropogon is one of the main economic fishes in Tibet area,only distributed in the upper and middle reaches of the Yarlung Tsangpo River and its tributaries.Saprolegniasis is one of the most difficult fish diseases in prevention and control,and brings huge economic losses to the aquaculture industry.This experiment was conducted to analyze the effects of Saprolegniasis on the spleen transcriptome of S.macropogon and understand the molecular mechanisms of S.macropogon infected by Saprolegniasis.A total of 5 healthy and 5 Saprolegnia infected S.macropogon living in the same aquatorium were randomly selected.The PDA agar medium and molecular biology methods were used for isolation and identification the S.strain from S.Macropogon.In addition to the transcriptomes of spleen tissues of healthy and fungus infection,S.macropogon were sequenced based on the Illumina HiSeqTM 2000 high-throughput sequencing platform and the data were analyzed by bioinformatics tools in this study.The research showed that we obtained 3 S.strains from the skin of S.Macropogon,including S.parasitica and S.ferax.A total of 46619504 and 43912876 clean reads were obtained in the healthy group and fungus infection group,respectively.359164 transcripts were assembled and 123863 unigenes were spliced after removing redundancy.Compared with the healthy group,1889 unigenes showed differential expressions in the fungus infection group,1414 unigenes were up-regulated and 475 unigenes were down-regulated among these genes.Six differentially expressed unigenes(natural killer cells-lysin gene,somatostatin gene,preproinsulin gene,chymotrypsin gene,protein disulfide isomerase gene and prolactin receptor gene)were randomly selected for quantitative real-time PCR and the results were consistent with the RNA-seq.The results of GO functional enrichment showed that the up-regulated genes are mainly enriched in 241 terms,and down-regulated genes are mainly enriched in 60 terms.These genes involve molecular functions,cell components and biological pr

关 键 词：巨须裂腹鱼 水霉菌 转录组测序 脾脏组织 差异表达基因 

分 类 号：S941.4[农业科学—水产养殖]