高温胁迫下罗非鱼源无乳链球菌的比较转录组分析

doi:10.16431/j.cnki.1671-7236.2023.06.004

摘要/Abstract

摘要： 【目的】从分子水平了解高温季节罗非鱼无乳链球菌病容易暴发的原因。【方法】在35和28 ℃分别培养无乳链球菌，提取总RNA并进行转录组测序；利用华大基因Dr.Tom自主分析平台进行差异表达基因分析，并针对差异基因进行了KEGG通路和GO功能的分类和富集；通过实时荧光定量PCR随机检测10个基因的差异表达，确定转录组数据的可靠性。【结果】共得到35和28 ℃培养条件下的显著性差异表达基因357个，其中上调基因229个，下调基因128个；GO数据库富集能得到多个具有显著性的功能集，多集中在碳源代谢相关的途径，包括碳水化合物分解代谢过程（carbohydrate catabolic process）、糖原代谢过程（glycogen metabolic process）、细胞葡聚糖代谢过程（cellular glucan metabolic process）等；差异基因中发现了多个与无乳链球菌致病性相关的基因，如烷基过氧化氢还原酶亚基、cAMP因子、C5a肽酶等编码基因；实时荧光定量PCR与转录组数据间皮尔逊相关系数R值为0.979（P<0.001），说明转录组数据可靠有效。【结论】高温能够显著影响无乳链球菌碳源代谢相关的途径，可能通过加快菌体利用碳源而促进菌体的生长和扩散。高温也诱导了多个与致病性相关的基因出现上调表达，其中cAMP因子引起的细胞膜溶解和氧化还原酶类引起的免疫逃避可能是高温下无乳链球菌致病性增强的主要因素。

关键词: 罗非鱼; 无乳链球菌; 转录组测序; cAMP因子; 烷基过氧化氢还原酶

Abstract: 【Objective】 The purpose of this study was to understand the causes of the outbreak of Streptococcus agalactis (S.agalactis) disease in high temperature season at the molecular level.【Method】 S.agalactis was cultured at 35 and 28 ℃ respectively, and total RNA was extracted for transcriptome sequencing.The differentially expressed genes(DEGs) were analyzed by Dr Tom's analysis platform of BGI, and the classification and enrichment of KEGG pathway and GO function were carried out.The reliability of transcriptome data was determined by randomly detecting the differential expression of 10 genes by Real-time quantitative PCR.【Result】 A total of 357 DEGs were obtained under 35 and 28 ℃, including 229 up-regulated genes and 128 down-regulated genes.The enrichment of GO database could obtain multiple functional sets with significant, most of which focus on the pathways related to carbon source metabolism, such as carbohydrate catabolic process, glycogen metabolic process, cellular glucan metabolic process, etc.. Many genes related to the pathogenicity of S.agalactis were found among the DEGs, such as the coding genes of alkyl hydroperoxide reductase subunit, CAMP factor, C5a peptidase, etc.. The Pearson correlation coefficient R value between Real-time quantitative PCR and transcriptome data was 0.979 (P<0.001), indicating that the transcriptome data was reliable and effective.【Conclusion】 High temperature could significantly affect the pathway related to the carbon source metabolism of S.agalactis, which might promote the growth and diffusion of the cell by accelerating the use of carbon source.High temperature also induced the up-regulation of several genes related to pathogenicity.Among them, cell membrane lysis caused by cAMP factor and immune escape caused by oxidoreductases might be the main factors that enhance the pathogenicity of S.agalactis under high temperature.

Key words: tilapia; Streptococcus agalactiae; transcriptome sequencing; cAMP factor; alkyl hydrogen peroxide reductase

中图分类号:

S852

施金谷, 黄珊珊, 黄基伟, 陈坤海, 胡大胜, 朱志雄, 韩书煜. 高温胁迫下罗非鱼源无乳链球菌的比较转录组分析[J]. 中国畜牧兽医, 2023, 50(6): 2207-2216.

SHI Jingu, HUANG Shanshan, HUANG Jiwei, CHEN Kunhai, HU Dasheng, ZHU Zhixiong, HAN Shuyu. Comparative Transcriptome Analysis of Streptococcus agalactiae from Tilapia Under High Temperature Stress[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2207-2216.

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