许氏平鲉对慢性高温胁迫响应的转录组学分析
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1.中国水产科学研究院黄海水产研究所, 海水养殖生物育种与可持续产出全国重点实验室, 农业农村部海洋渔业可持续发展重点实验室, 山东 青岛 266071 ;2.烟台市海洋经济研究院, 山东 烟台 264006

作者简介:

谭杰(1980-),男,博士,助理研究员,研究方向为水产养殖学.E-mail:tanjie@ysfri.ac.cn

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S917

基金项目:

山东省重点研发计划项目(科技示范工程)项目(2021SFGC0701); 中国水产科学研究院黄海水产研究所基本科研业务费项目(20603022024026)


Transcriptomic responses to chronic heat stress in black rockfish (Sebastes schlegelii)
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1.State Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Key Laboratory of Marine Fisheries andSustainable Development, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences, Qingdao 266071 , China ;2.Yantai Marine Economy Institute, Yantai 264006 , China

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    摘要:

    为探索许氏平鲉(Sebastes schlegelii)高温胁迫条件下基因表达水平的变化, 利用 Illumina Hiseq 4000 测序平台分别对许氏平鲉高温处理组(温度由 18 ℃缓慢升至 27 ℃后养殖 5 d)和常温对照组(18 ℃下养殖)的鳃、肝和肠道组织进行了转录组测序。以|log2fold change|≥1 和 FDR (false discovery rate, 错误发现率)<0.05 作为显著差异表达基因(DEGs)筛选条件。在鳃组织中, 共筛选出 306 个 DEGs, 其中 96 个基因显著上调, 210 个基因显著下调; 在肝脏组织中, 共筛选出 806 个 DEGs, 其中 382 个基因显著上调, 424 个基因显著下调; 在肠道组织中, 共筛选出 343 个 DEGs, 其中 162 个基因显著上调, 181 个基因显著下调。3 个组织共表达的 DEGs 有 12 个。GO 功能注释分析结果显示, 鳃组织中的 DEGs 较多富集在蛋白水解作用、胞外区、结构分子活性、受体调节器活性、受体配体活性等功能中; 肝组织中 DEGs 较多富集在脂肪代谢过程、细胞氨基酸代谢过程、细胞质、氧化还原酶活性等功能中; 肠道组织中 DEGs 较多富集在氧化还原过程、胞外区、辅因子结合等功能中。运用逆转录实时荧光定量 PCR (RT-qPCR) 对 10 个 DEGs 的表达量进行验证, 结果显示, RT-qPCR 结果与转录组分析结果一致, 表明转录组分析可靠。本研究为解析许氏平鲉对慢性高温胁迫应激反应的分子机制提供了转录水平上的参考数据。

    Abstract:

    Black rockfish (Sebastes schlegelii) inhabit the coastal regions of the northwestern Pacific Ocean, including the East China Sea, Yellow Sea, and the coastal areas of Korea and Japan. It is highly favored by consumers because of its delicate flavor and high nutritional content. Black rockfish is among the most economically cultivated marine species in northern China. The juveniles were raised in offshore net cages. The optimum temperature for black rockfish growth ranges from 18 ℃ to 24 ℃. In summer, the seawater temperature sometimes exceeds 28 ℃ in black rockfish farms. As global warming intensifies, long-term exposure to high temperatures in summer will affect the growth and survival of black rockfish cultured in net cages, indicating that the effects of chronic heat stress on black rockfish merit further research. However, the molecular mechanisms underlying the responses of black rockfish to chronic heat stress remain largely unknown. Understanding these mechanisms will improve fish welfare and farm production. Marker genes to monitor heat stress are required to identify heat-resistant fish. In this study, we conducted RNA-seq analysis to characterize the genes and pathways involved in chronic thermal stress responses in the gills, liver, and intestines of black rockfish. Healthy black rockfishes were cultured at a normal temperature (18 ℃) and a high temperature (27 ℃). For the chronic heat stress treatment, water temperature was increased from 18 ℃ to 27 ℃ at a constant rate of 1 ℃ per day, and maintained for 10 days. The gill, liver, and intestinal tissues were used as experimental materials in both the heat stress and normal groups. Total RNA was extracted, and 18 mRNA libraries were constructed and sequenced using the Illumina HiSeq-4000 technology platform. Differentially expressed genes were analyzed using edgeR. Bioinformatic analysis was performed on the GO and KEGG functions of the differentially expressed genes, and the key differentially expressed genes were further validated using RT-qPCR. In total, 306 annotated differentially expressed genes (DEGs) were identified in the gill, of which 96 and 210 were up-and downregulated, respectively. In total, 806 annotated differentially expressed genes (DEGs) were identified in the liver, of which 382 were upregulated and 424 were downregulated. A total of 343 annotated differentially expressed genes (DEGs) were identified in the intestine, among which 162 and 181 were up-and downregulated, respectively. A Venn diagram showed that 12 DEGs were shared among three tissues. And 40, 53, and 49 DEGs were shared between liver and gill, liver and intestine, gill and intestine, respectively. Furthermore, GO functional enrichment analysis revealed that the DEGs were mainly enriched in proteolysis, extracellular region, structural molecule activity, receptor regulator activity, and receptor-ligand activity in the gill; in lipid metabolic process, cellular amino acid metabolic process, cytoplasm, and oxidoreductase activity in the liver; and in oxidation-reduction process, extracellular region, and cofactor binding in the intestine. Among the up-regulated genes under heat stress included the heat shock proteins 90α, period circadian protein homolog 2, serpin H1, and the down-regulated genes included period circadian protein homolog 1. Ten DEGs were subjected to reverse transcription quantitative PCR (RT-qPCR) for relative quantification to assess the differences in gene expression between the normal and high temperature groups. The expression trends observed in the RT-qPCR analysis were consistent with those identified in the RNAseq data, which confirmed the reliability of the transcriptomic sequencing results. These results provide abundant data for further studies on the molecular mechanisms of the chronic heat stress response in Sebastes schlegelii.

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谭杰,张正,刘长琳,王亮,肖文杰,姜达.许氏平鲉对慢性高温胁迫响应的转录组学分析[J].中国水产科学,2024,31(12):1453-1463
TAN Jie, ZHANG Zheng, LIU Changlin, WANG Liang, XIAO Wenjie, JIANG Da. Transcriptomic responses to chronic heat stress in black rockfish (Sebastes schlegelii)[J]. Journal of Fishery Sciences of China,2024,31(12):1453-1463

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  • 收稿日期:2024-07-19
  • 最后修改日期:2024-09-02
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  • 在线发布日期: 2025-02-11
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