Transcriptomic analysis of Pacific white shrimp (Litopenaeus vannamei, Boone 1931) in response to acute hepatopancreatic necrosis disease caused by Vibrio parahaemolyticus
Transcriptomic analysis of Pacific white shrimp (Litopenaeus vannamei, Boone 1931) in response to acute hepatopancreatic necrosis disease caused by Vibrio parahaemolyticus
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DOI：
10.1371/journal.pone.0220993
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发表时间：
2019-08-13
期刊：
影响因子：
3.7
通讯作者：
Ascencio, Felipe
中科院分区：
文献类型：
--
作者：
Velazquez-Lizarraga, Adrian E.;Luis Juarez-Morales, Jose;Ascencio, Felipe
由海洋细菌副溶血性弧菌引起的急性肝胰腺坏死病（AHPND）是对虾养殖中的一个严重问题。副溶血性弧菌感染物质包含在编码致死毒素PirAB（Vp）的质粒中，其主要靶组织是肝胰腺，引起上皮细胞脱落、坏死和大量血细胞浸润。为了更好地了解AHPND期间肝胰腺的反应，通过浸泡副溶血性弧菌感染凡纳滨对虾幼虾。在感染后24小时，我们对感染的虾肝胰腺进行了转录组mRNA测序，以鉴定新的差异表达基因，共检测了174，098个转录本，其中915个转录本在比较转录组分析后发现差异表达：442个上调转录本和473个下调转录本。基因本体论术语富集分析上调转录物包括代谢过程、程序性细胞死亡的调节、碳水化合物代谢过程和生物粘附，而下调转录物包括基于微管的过程、细胞活化和几丁质代谢过程。对上调和下调基因之间蛋白质网络的分析表明，第一个基因相互作用与氧化过程和肌节组织有关。此外，蛋白质-蛋白质网络分析确定了20个顶部高度连接的枢纽节点。根据其免疫或代谢功能，选择10个候选转录子，采用RT-qPCR方法检测其在AHPND感染的对虾肝胰腺中的mRNA相对表达水平。我们的研究结果表明，在AHPND感染的免疫和代谢系统之间的密切联系。我们的RNA-Seq和RT-qPCR数据提供了可能的免疫学和生理学场景以及在虾肝胰腺中响应感染性疾病的分子途径。
Acute hepatopancreatic necrosis disease (AHPND), caused by marine bacteria Vibrio Parahaemolyticus, is a huge problem in shrimp farms. The V. parahaemolyticus infecting material is contained in a plasmid which encodes for the lethal toxins PirAB(Vp), whose primary target tissue is the hepatopancreas, causing sloughing of epithelial cells, necrosis, and massive hemocyte infiltration. To get a better understanding of the hepatopancreas response during AHPND, juvenile shrimp Litopenaeus vannamei were infected by immersion with V. parahaemolyticus. We performed transcriptomic mRNA sequencing of infected shrimp hepatopancreas, at 24 hours post-infection, to identify novel differentially expressed genes a total of 174,098 transcripts were examined of which 915 transcripts were found differentially expressed after comparative transcriptomic analysis: 442 up-regulated and 473 down-regulated transcripts. Gene Ontology term enrichment analysis for up-regulated transcripts includes metabolic process, regulation of programmed cell death, carbohydrate metabolic process, and biological adhesion, whereas for down-regulated transcripts include, microtubule-based process, cell activation, and chitin metabolic process. The analysis of protein-protein network between up and down-regulated genes indicates that the first gene interactions are connected to oxidation-processes and sarcomere organization. Additionally, protein-protein networks analysis identified 20-top highly connected hub nodes. Based on their immunological or metabolic function, ten candidate transcripts were selected to measure their mRNA relative expression levels in AHPND infected shrimp hepatopancreas by RT-qPCR. Our results indicate a close connection between the immune and metabolism systems during AHPND infection. Our RNA-Seq and RT-qPCR data provide the possible immunological and physiological scenario as well as the molecular pathways that take place in the shrimp hepatopancreas in response to an infectious disease.