嗜水气单胞菌感染后唇䱻的转录组测序及相关免疫基因分析  

Transcriptome Sequencing and Analysis of Immune-related Genes of Barbel Steed(Hemibarbus labeo)After Aeromonas hydrophila Infection

作　　者：陈洁[1,2,3] 俞静 朱群茵 刘子明 陈旭堂[3] 朱阳春 丁国骅 舒妙安[2] CHEN Jie;YU Jing;ZHU Qun-Yin;LIU Zi-Ming;CHEN Xu-Tang;ZHU Yang-Chun;DING Guo-Hua;SHU Miao-An(College of Ecology,Lishui University,Lishui 323000,China;College of Animal Sciences,Zhejiang University,Hangzhou 310058,China;Lishui Institute for Ecological Economy Research,Lishui 323000,China)

机构地区：[1]丽水学院生态学院,丽水323000 [2]浙江大学动物科学学院,杭州310058 [3]丽水市生态经济研究所,丽水323000

出　　处：《农业生物技术学报》2023年第1期136-145,共10页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(32002407);丽水市重点研发计划项目(2022ZDYF02);丽水市公益性技术研究项目(2022GYX22)。

摘　　要：细菌性肠炎在水产养殖过程中频繁发生,然而细菌性肠炎的确切机制尚不十分清楚。本研究旨在鉴定与嗜水气单胞菌(Aeromonas hydrophila)感染诱导的肠炎相关的新候选基因,以嗜水气单胞菌感染唇䱻(Hemibarbus labeo)为材料,利用BGISEQ-500平台,对其肠道进行转录组测序。结果显示,测序共获得120173个unigene,平均长度为1328 bp,组装质量指标N50为2647 bp,其中72.71%为已知基因。差异表达基因筛选发现,嗜水气单胞菌感染后共有33706个基因显著差异表达,其中18878个基因上调,14828个基因下调。通过GO富集分析发现,差异表达基因主要富集于细胞过程、代谢进程、生物调控、细胞、细胞组分和结合等。通过KEGG富集分析发现,差异表达基因主要富集于信号转导、免疫系统、癌症和传染病等信号通路。差异表达基因分析和qPCR验证显示,抗菌肽基因铁调素(hepcidin)、肝表达抗菌肽2(liver-expressed antimicrobial peptide 2,leap2)和炎症细胞因子基因白细胞介素1β(interleukin-1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、IL6和IL8的表达在嗜水气单胞菌感染后显著上调。化学合成Hepcidin、LEAP2和细胞溶素(NK-lysin)并检测了其对嗜水气单胞菌的抗菌活性,发现Hepcidin和LEAP2可以有效抑制嗜水气单胞菌的繁殖。本研究为进一步了解唇䱻和其他养殖鱼类细菌性肠炎的致病机制提供了基础数据。Bacterial enteritis occurs frequently in aquaculture,however,the exact mechanism of bacterial enteritis was not well understood.This study aimed to identify novel candidate genes associated with Aeromonas hydrophila infection-induced enteritis.Barbel steed(Hemibarbus labeo)were infected with A.hydrophila,transcriptome sequencing of its intestine using the BGISEQ-500 platform.The results showed that a total of 120173 unigenes were obtained by sequencing,with an average length of 1328 bp and an N50 of2647 bp,of which 72.71%were known genes.Screening of differentially expressed genes(DEGs)found that there were 33706 genes significantly differentially expressed after A.hydrophila infection,of which 18878unigenes were up-regulated,and 14828 genes were down-regulated.Through GO enrichment analysis,DEGs were mainly enriched in cellular process,metabolic process,biological regulation,cell,cell part,and binding.The KEGG enrichment analysis showed that DEGs were mainly enriched in signal pathways such as signal transduction,immune system,cancers,infectious diseases,etc.According to DEGseq analysis and qPCR verification,it was found that the antimicrobial peptide genes,antimicrobial peptide genes(hepcidin and liverexpressed antimicrobial peptide 2(leap2))and inflammatory cytokine genes(interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),IL6 and IL8)were significantly increased after A.hydrophila infection.Meanwhile,the antibacterial activities of chemically synthesized Hepcidin,LEAP2 and NK-lysin against A.hydrophila were detected,and it was found that Hepcidin and LEAP2 could effectively inhibit the reproduction of A.hydrophila.This study provides basic data for further understanding of the pathogenic mechanism of bacterial enteritis in barbel steed and other farmed fish.

关 键 词：唇䱻 嗜水气单胞菌 转录组分析 抗菌肽 炎症因子 

分 类 号：S917.4[农业科学—水产科学]