南方农业学报2025，Vol.56Issue(6)：1752-1762,11.DOI:10.3969/j.issn.2095-1191.2025.06.006
基于转录组测序的花斑裸鲤温度应激功能基因挖掘
Mining of temperature stress functional genes in Gymnocypris eckloni based on transcriptome sequencing
摘要
Abstract
[Objective]The purpose of this study was to explore the temperature stress functional genes of Gymnocy-pris eckloni based on transcriptome sequencing,which could provide theoretical reference for exploring the adaptability and temperature response mechanism of Gymnocypris eckloni under heat stress.[Method]Sixty healthy Gymnocypris eck-loni were selected and randomly divided into a control group and a heat stress group,which were reared in 10 and 28℃water environments respectively.After the rearing,the liver tissues were obtained after MS-222 anesthesia.The liver tis-sues were ground with liquid nitrogen and then the total RNA was extracted by TRIzol method.The high-throughput se-quencing was performed using the Illumina NovaSeq 6000 platform.The obtained unigenes were compared with the NR,NT,KO,SWISS-PROT,Pfam,GO and KOG databases for functional annotation analysis.Based on the transcript ex-pression level,differentially expressed genes(DEGs)were screened by DESeq2,and KEGG signaling pathway enrich-ment analysis was performed.Six DEGs were randomly selected for real-time fluorescence quantitative PCR detection to verify the accuracy of the transcriptome data.[Result]GO functional annotation analysis results showed that a total of 23464 unigenes were successfully annotated in the GO database,involving 3 functional categories:biological processes,cellular components and molecular functions.KOG functional annotation analysis results showed that 16583 unigenes were successfully annotated in the KOG database,involving 26 KOG functional terms such as RNA processing and modi-fication of chromosome structure and dynamics.KEGG functional annotation analysis results showed that unigenes anno-tated to the KEGG database involved in 5 major functional categories,among which the KEGG functional term with the largest number of annotated unigenes was signal transduction,followed by the immune system.DEGs analysis results showed that there were 2388 DEGs between the control group and the heat stress group,1520 DEGs were up-regulated,and 868 DEGs were down-regulated.KEGG signaling pathway enrichment analysis results showed that the up-regulated DEGs were mainly enriched in signaling pathways such as antigen processing and presentation,endoplasmic reticulum protein processing,and estrogen.The down-regulated DEGs mainly enriched in metobolic pathways.The results of real-time fluorescence quantitative PCR showed that fragments per kilobase of exon model per million mapped fragments(FPKM)in DNAJA4,HSP90a,FKBP4,Caspase1,Caspase7 and Caspase8 transcriptomes were consistent with the gene expression variation trend,indicating that the transcriptome sequencing data had a high accuracy.[Conclusion]Under heat stress,the expression levels of immune-related DEGs of Gymnocypris eckloni are down-regulated,and the expres-sion levels of heat shock protein are up-regulated.Heat stress may inhibit the immune function of Gymnocypris eckloni,and the heat shock protein family members may play an anti-heat stress role by stabilizing protein structure and maintai-ning cell homeostasis.Under heat stress,the expression levels of apoptosis-related DEGs Caspase7 and Caspase8 in Gym-nocypris eckloni are significantly down-regulated,which may be related to the inhibitory effect of heat shock protein on apoptosis.Gymnocypris eckloni may reduce the risk of apoptosis by enhancing the activity of molecular chaperones under heat stress.关键词
花斑裸鲤/热应激/细胞凋亡/热休克蛋白Key words
Gymnocypris eckloni/heat stress/cell apoptosis/heat shock protein分类
农业科技引用本文复制引用
吴晓雲,陈叶雨,刘亚,宋明江,李飞扬,邹巧林,赖见生..基于转录组测序的花斑裸鲤温度应激功能基因挖掘[J].南方农业学报,2025,56(6):1752-1762,11.基金项目
农业农村部财政专项"黄河渔业资源与环境调查"(HHDC-2025-0104) （HHDC-2025-0104）
国家现代农业产业技术体系四川淡水鱼创新团队建设专项(SCCXTD-2025-15) （SCCXTD-2025-15）
四川省科技计划项目(20211YFYZ0015) The Project of Yellow River Fisheries Resources and Environment Investigation from the MARA,P.R.China(HHDC-2025-0104) （20211YFYZ0015）
Sichuan Freshwater Fish Innovation Team Construction Project of China Agri-culture Research System(SCCXTD-2025-15) （SCCXTD-2025-15）
Sichuan Science and Technology Plan Project(20211YFYZ0015) （20211YFYZ0015）
