Differentially Expressed Genes and Alternative Splicing Analysis Revealed the Difference in Virulence to American Eels (Anguilla rostrata) Infected by Edwardsiella anguillarum and Aeromonas hydrophila

IF 2.6 3区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Peng Lin, Zihao Chen, Guanghua Sun, Songlin Guo
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引用次数: 0

Abstract

Edwardsiella anguillarum and Aeromonas hydrophila are two common bacterial pathogens affecting cultivated eels, and the differences in their virulence remain unclear. In this study, after two groups of American eels (Anguilla rostrata) were administered the LD50 dose of E. anguillarum and A. hydrophila, respectively, the histopathology of the liver, trunk kidney, and spleen, as well as transcriptomic RNA sequencing (RNA-seq) analysis of the spleen, was examined at three time points: pre-infection (Con group) and post-infection at 36 h (Ea_36 group, Ah_36 group) and 60 h (Ea_60 group, Ah_60 group). The results showed that the differences in pathological changes were characterized by severe hepatocyte edema at 36 h post-infection (hpi) and hepatocyte atrophy at 60 hpi in the livers of eels infected by A. hydrophila, in contrast to the severe atrophy of glomeruli in the trunk kidneys and numerous bacterial nodules in the spleens of eels infected by E. anguillarum. The RNA-seq results revealed 906 and 77 typical differentially expressed genes (DEGs) in eels infected with E. anguillarum and A. hydrophila, respectively, compared to the control eels. The DEGs between the infected and control groups were predominantly annotated in GO terms related to binding, catalytic activity, membrane part, cell part, and cellular process, as well as in KEGG pathways associated with human diseases and organismal systems. The GO enrichment analysis showed 83 and 146 differential GO terms, along with 32 and 78 differential KEGG pathways in two comparisons of Ea_36 vs Con versus Ah_36 vs Con and Ea_60 vs Con versus Ah_60 vs Con, respectively. Furthermore, the analysis of differential alternative splicing genes (DASs) showed 1244 and 1341 DASs out of 12,907 and 12,833 AS genes, respectively, in the comparisons of Ea_36 vs Ah_36 and Ea_60 vs Ah_60. These DASs were enriched in two common KEGG pathways: “NOD-like receptor signaling pathway” and “necroptosis” which shared 11 hub DASs. Finally, analysis of protein–protein interactions revealed that 91 of 412 cross DASs between Ea_36 vs Ah_36 and Ea_60 vs Ah_60 potentially play an essential role in the difference in virulence of E. anguillarum and A. hydrophila in American eels, with 12 encoded proteins being particularly notable. Together, this study is the first to report a comparative pathogenicity and RNA-seq analysis of E. anguillarum and A. hydrophila in American eels, shedding new light on our understanding of the differences in virulence as revealed by pathological changes, DEGs, and DASs, contributing to more effective control strategies to prevent outbreaks of bacterial infections.

差异表达基因和交替剪接分析揭示了鳗鲡和嗜水气单胞菌感染美洲鳗的毒性差异。
鳗鲡爱德华氏菌(Edwardsiella anguillarum)和嗜水气单胞菌(Aeromonas hydrophila)是影响养殖鳗鱼的两种常见细菌病原体,它们的致病力差异仍不清楚。在本研究中,两组美洲鳗鲡分别摄入 LD50 剂量的鳗鲡嗜血杆菌和嗜水气单胞菌后,其组织结构发生了变化。分别在感染前(Con 组)、感染后 36 小时(Ea_36 组,Ah_36 组)和 60 小时(Ea_60 组,Ah_60 组)三个时间点检测了肝脏、肾脏和脾脏的组织病理学以及脾脏的转录组 RNA 测序(RNA-seq)分析。结果表明,感染鳗鲡肝脏后 36 h 肝细胞水肿严重,60 h 肝细胞萎缩,而感染鳗鲡肾脏后躯干肾小球严重萎缩,脾脏内有大量细菌结节,两者的病理变化存在差异。RNA-seq结果显示,与对照组相比,感染鳗鲡和纤毛虫的鳗鱼分别有906个和77个典型的差异表达基因(DEGs)。感染组和对照组之间的差异表达基因主要被注释在与结合、催化活性、膜部分、细胞部分和细胞过程相关的GO术语中,以及与人类疾病和生物系统相关的KEGG通路中。GO富集分析显示,在Ea_36 vs Con与Ah_36 vs Con和Ea_60 vs Con与Ah_60 vs Con的两次比较中,分别出现了83和146个不同的GO术语,以及32和78个不同的KEGG通路。此外,对差异替代剪接基因(DAS)的分析表明,在 Ea_36 vs Ah_36 和 Ea_60 vs Ah_60 的比较中,12907 和 12833 个 AS 基因中分别有 1244 和 1341 个 DAS。这些DASs富集在两个共同的KEGG通路中:"NOD样受体信号通路 "和 "坏死 "共享11个枢纽DAS。最后,蛋白质-蛋白质相互作用分析表明,在 Ea_36 与 Ah_36 之间以及 Ea_60 与 Ah_60 之间的 412 个交叉 DAS 中,有 91 个可能在鳗鲡和嗜水杆线虫在美洲鳗鱼中的毒力差异中发挥重要作用,其中 12 个编码蛋白质尤为显著。总之,本研究首次报告了鳗鲡和纤毛虫在美洲鳗鱼中的致病性比较和 RNA-seq 分析,为我们了解病理变化、DEGs 和 DASs 揭示的致病性差异提供了新的思路,有助于制定更有效的控制策略,防止细菌感染的爆发。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Marine Biotechnology
Marine Biotechnology 工程技术-海洋与淡水生物学
CiteScore
4.80
自引率
3.30%
发文量
95
审稿时长
2 months
期刊介绍: Marine Biotechnology welcomes high-quality research papers presenting novel data on the biotechnology of aquatic organisms. The journal publishes high quality papers in the areas of molecular biology, genomics, proteomics, cell biology, and biochemistry, and particularly encourages submissions of papers related to genome biology such as linkage mapping, large-scale gene discoveries, QTL analysis, physical mapping, and comparative and functional genome analysis. Papers on technological development and marine natural products should demonstrate innovation and novel applications.
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