基于转录组学分析黑斑侧褶蛙歪头病机理  

Based on Transcriptomic Analysis of the Mechanism of the Crooked Head Disease in the Pelophylax nigromaculata

作　　者：任思齐 蔡卓 郑安瑞 叶嘉政 佟皖晴 李倩慧 郑善坚[1,2] REN Si-Qi;CAI Zhuo;ZHENG An-Rui;YE Jia-Zheng;TONG Wan-Qing;LI Qian-Hui;ZHENG Shan-Jian(College of Bioscience,Zhejiang Normal University,Jinhua 321004,China;Key Laboratory of Wildlife Biotechnology and Conservation and Utilization of Zhejiang Province,Jinhua 321004,China)

机构地区：[1]浙江师范大学生命科学学院,金华321004 [2]浙江省野生动物生物技术与保护利用重点实验室,金华321004

出　　处：《农业生物技术学报》2024年第12期2822-2836,共15页Journal of Agricultural Biotechnology

基　　金：浙江省重点研发计划(2021C02044)。

摘　　要：歪头病是黑斑侧褶蛙(Pelophylax nigromaculata)养殖中危害最大的病症之一,该病死亡率高、传染性强,给蛙类养殖造成了巨大损失,且目前对该病无有效防治手段。针对目前黑斑侧褶蛙的养殖现状,急需探明歪头病的致病机理,以减少黑斑侧褶蛙的养殖风险。本研究对12只患歪头病的黑斑侧褶蛙进行病原菌分离,得到1株优势菌HBG0512,取患病和健康黑斑侧褶蛙的脑及肝脏组织进行转录组测序。获得了差异表达基因,对其进行GO和KEGG分析,并采用实时荧光定量PCR验证基因表达水平。研究结果显示,从患病黑斑侧褶蛙分离的HBG0512菌株为米尔伊丽莎白菌(Elizabethkingia miricola);转录组测序发现患病与健康黑斑侧褶蛙的脑与肝脏组织各有212和612个差异基因;GO富集分析发现了肝脏内质网相关基因的异常表达及脑组织促炎因子的积累;KEGG通路分析揭示,导致内质网应激的激活转录因子6(activating transcription factor 6,ATF6)异常表达,脑部炎症反应则可能是由IgE/FcεRI通路介导发生。患歪头病黑斑侧褶蛙的肝脏及脑部均有炎症反应发生,肝脏内质网感受器ATF6基因显著下调,内质网折叠反应能力下降,未折叠蛋白聚集导致了内质网应激,致使炎症反应发生;脑部主要通过IgE/FcεRI介导的炎症反应,导致血脑屏障通透性破坏,引起细菌性歪头病的发生。本研究通过转录组学揭示了黑斑侧褶蛙歪头病的致病机理,为蛙类及其他两栖爬行动物的疾病防治提供参考。Crooked head disease is one of the most harmful diseases in Pelophylax nigromaculata breeding.It has high mortality and strong infectivity.This disease has a high mortality rate and strong infectivity,causing significant losses in frog farming.Currently,there are no effective prevention or treatment methods for this disease.In view of the current breeding status of P.nigromaculata,it is necessary to explore the pathogenic mechanism of the crooked head disease in order to reduce the breeding risks of P.nigromaculata.In this study,the aseptic operation of P.nigromaculata suffering from the crooked head disease was inoculated with bacteria,and a dominant bacteria HBG0512 was isolated.The brain and liver tissues of diseased and healthy black spotted frogs were sequenced.The differentially expressed genes were obtained and analyzed by GO and KEGG,and the gene expression level was verified by real-time fluorescence quantitative PCR.The results showed that the HBG0512 strain isolated from the diseased P.nigromaculata was Elizabethkingia miricola.Transcriptome sequencing found 212 and 612 differential genes in the brain and liver tissues of the diseased and healthy P.nigromaculata.GO enrichment analysis showed abnormal expression of liver endoplasmic reticulum related genes and accumulation of pro-inflammatory factors in brain tissue.KEGG pathway analysis revealed that activating transcription factor 6(ATF6)was abnormally expressed,which caused endoplasmic reticulum stress,and brain inflammatory response might be mediated by the IgE/FcεRI pathway.Inflammation occured in the liver and brain of P.nigromaculata with the crooked head disease,the liver endoplasmic reticulum receptor ATF6 gene was significantly down-regulated,the ability of endoplasmic reticulum folding reaction was decreased,and the accumulation of unfolded proteins led to endoplasmic reticulum stressinduced inflammation;brain inflammation was mainly mediated by IgE/FcεRI,which led to the destruction of blood-brain barrier permeability and the occurrence of

关 键 词：歪头病 黑斑侧褶蛙 转录组 内质网应激 激活转录因子6(ATF6) 

分 类 号：S917.4[农业科学—水产科学]