不同培养温度的鱼源海豚链球菌转录组分析  

Transcriptome analysis of Streptococcus iniae from fish at different culture temperatures

作　　者：贾新蕾 黄增朝 杨林狄 吕静 李妍萍 简纪常[1] 黄郁葱[1,2] JIA Xinlei;HUANG Zengchao;YANG Lindi;LYU Jing;LI Yanping;JIAN Jichang;HUANG Yucong(Fisheries College of Guangdong Ocean University&Guangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Aquaculture,Zhanjiang,Guangdong 524088,China;Southern Marine Science and Engineering Guangdong Laboratory(Zhanjiang),Zhanjiang,Guangdong 524006,China)

机构地区：[1]广东海洋大学水产学院/广东省水产动物病害防控与健康养殖重点实验室,广东湛江524088 [2]南方海洋科学与工程广东省实验室(湛江),广东湛江524006

出　　处：《热带生物学报》2024年第1期109-121,共13页Journal of Tropical Biology

基　　金：湛江市科技计划项目(2021A05196);国家重点研发计划项目(2022YFD2401200);广东省重点领域研发计划项目(2021B0202040002);南方海洋科学与工程广东省实验室(湛江)资助项目(ZJW-2019-06)。

摘　　要：为鉴定海豚链球菌(Streptococcus iniae)在不同温度的转录水平差异,通过生长曲线测定和人工感染试验分析该菌在25℃和35℃下的生长情况和致病性,采用链特异性转录组测序(Strand-specific RNA-seq)技术对25℃和35℃培养的海豚链球菌Tozj-1菌株进行测序分析,筛选差异表达基因,通过GO(Gene Ontology)数据库和KEGG(Kyoto Encyclopedia of Genes and Genomes)数据库对差异表达基因进行GO功能和KEGG通路富集分析,然后利用VFDB(Virulence factor database)数据库筛选差异表达的重要毒力基因并使用实时荧光定量PCR进行验证。结果显示,海豚链球菌在35℃条件下有更快的生长速度和更强的毒力;转录组共筛选获得927个显著差异表达基因(P<0.05),其中包括820个上调基因和107个下调基因;GO功能富集分析发现,差异基因主要富集在代谢、细胞、催化及结合过程;KEGG富集分析发现,差异基因主要富集在核糖体通路、ABC转运通路、群体感应等信号通路。以上结果表明温度调控海豚链球菌基因的转录表达及相关通路的富集,为后续海豚链球菌致病机理的研究提供数据支持。To identify the differences in transcription level of Streptococcus iniae at different temperatures,the growth and pathogenicity of S.iniae cultured at 25℃and 35℃were analyzed by using growth curve and artificial infection test,and strand-specific RNA-seq technology was used for sequencing analysis of S.iniae Tozj-1 strain cultured at 25℃and 35℃.The differentially expressed genes(DEGs)were screened and their GO function and KEGG pathway enrichment were analyzed based on the GO(Gene Ontology)database and KEGG(Kyoto Encyclopedia of Genes and Genomes)database.The key differentially expressed virulence genes were filtrated using virulence factor database(VFDB)and verified by real-time quantitative PCR.The results showed that S.iniae grew faster with a higher virulence at 35℃.A total of 927 significantly differentially expressed genes were screened(P<0.05),including 820 up-regulated genes and 107 down-regulated genes.GO functional enrichment analysis showed that the DEGs were mainly enriched in metabolic process,cellular process,binding process and catalytic activity.KEGG enrichment analysis revealed that the DEGs were mainly enriched in ribosome,quorum sensing,ABC transporters and other signaling pathways.These results indicated that temperature regulated the transcription and expression of S.iniae genes and the enrichment of related pathways,which provided data support for further research in the pathogenesis of S.iniae.

关 键 词：海豚链球菌 转录组测序 温度 毒力因子 QRT-PCR 

分 类 号：Q819[生物学—生物工程]