生物技术通报 ›› 2024, Vol. 40 ›› Issue (11): 321-334.doi: 10.13560/j.cnki.biotech.bull.1985.2024-0184

• 研究报告 • 上一篇    下一篇

克氏原螯虾响应高温胁迫的生理代谢机制

邹永烽1,2(), 包志明1,2, 曹攀辉1,2, 张嘉媛1,2, 郭介宇1,2, 苏现斌1,2, 徐宇3, 许志强3, 郭慧1,2()   

  1. 1.广东海洋大学水产学院,湛江 524000
    2.湛江市海洋生态与养殖环境重点实验室,湛江 524088
    3.江苏省淡水水产研究所 农业农村部淡水虾蟹遗传育种与养殖重点实验室,南京210017
  • 收稿日期:2024-02-28 出版日期:2024-11-26 发布日期:2024-12-19
  • 通讯作者: 郭慧,女,副教授,研究方向:水产动物健康养殖生态学;E-mail: guoh@gdou.edu.cn
  • 作者简介:邹永烽,男,硕士研究生,研究方向:水产动物健康养殖;E-mail: zouyf111@126.com
  • 基金资助:
    江苏种业振兴“揭榜挂帅”项目(JBGS〔2021〕119)

Physiological and Metabolic Mechanisms of Procambarus clarkii in Response to High Temperature Stress

ZOU Yong-feng1,2(), BAO Zhi-ming1,2, CAO Pan-hui1,2, ZHANG Jia-yuan1,2, GUO Jie-yu1,2, SU Xian-bin1,2, XU Yu3, XU Zhi-qiang3, GUO Hui1,2()   

  1. 1. College of Fisheries, Guangdong Ocean University, Zhanjiang 524000
    2. Zhanjiang Key Laboratory of Marine Ecology and Aquaculture Environment, Zhanjiang 524088
    3. Key Laboratory of Genetic Breeding and cultivation for Freshwater Crustacean, Ministry of Agriculture and Rural Affairs, Freshwater Fisheries Research Institute of Jiangsu Province, Nanjing 210017
  • Received:2024-02-28 Published:2024-11-26 Online:2024-12-19

摘要:

【目的】 研究克氏原螯虾响应高温胁迫的生理代谢机制,为改善其养殖管理和品种选育提供参考。【方法】 以26℃为对照组,对克氏原螯虾进行32℃和37℃的高温胁迫实验,分别在胁迫后24 h和72 h收集鳃丝样品。通过组织学观察、生化指标测定(丙二醛、活性氧、总抗氧化能力、丙酮酸激酶、己糖激酶、碱性磷酸酶、磷酸烯醇式丙酮酸羧激酶、溶菌酶、酸性磷酸酶)和转录组学分析,综合探讨克氏原螯虾响应高温胁迫的生理代谢机制。【结果】 高温导致鳃丝组织损伤,生化指标总体上随着温度的升高而下降。与对照组相比,32℃胁迫组在24和72 h分别鉴定出226和6 937个差异表达基因(DEGs);37℃胁迫组则有高达2 928和9 449个DEGs。GO和KEGG富集分析表明,DEGs显著富集在O-聚糖生物合成、细胞外基质-受体相互作用、花生四烯酸代谢、鞘脂代谢以及淀粉和蔗糖代谢等过程。此外,加权基因表达网络分析发现DEGs聚类在免疫和能量代谢等模块上,并筛选出组蛋白(H2A)、线粒体(ND6)等核心基因。选取10个DEGs进行RT-qPCR验证,结果和测序结果一致,验证了转录组结果的准确性。【结论】 在32℃和37℃的温度下,克氏原螯虾的鳃丝受到损伤并降低了葡萄糖的利用率,迫使其通过调整热休克蛋白、抗氧化能力和代谢等途径维持细胞功能。

关键词: 克氏原螯虾, 高温, 鳃, 生理代谢, 机制

Abstract:

【Objective】 To understand the physiological and metabolic mechanisms of Procambarus clarkii in response to high temperature stress, and to provide references for improving P. clarkii culture management and variety breeding. 【Method】 The P. clarkii were exposed to 32℃ and 37℃, and 26℃ serving as the control condition. Gill samples were collected at 24 and 72 h post-exposure. The study employed a comprehensive analysis of histological observation, biochemical indicators(malondialdehyde, reactive oxygen species, total antioxidant capacity, pyruvate kinase, hexokinase, alkaline phosphatase, phosphoenolpyruvate carboxykinase, lysozyme, and acid phosphatase), and transcriptome to comprehensively explore the physiological and metabolic responses of P. clarkii to high temperature stress. 【Result】 High temperature damaged the gill tissues and the biochemical indicators overall decreased with the increasing of temperature. When compared to the control group, a total of 226 and 6 937 differentially expressed genes(DEGs)were identified at 24 and 72 h in 32℃ group, and a total of 2 928 and 9 449 DEGs were identified at 24 and 72 h in 37℃ group. GO and KEGG enrichment analyses indicate that DEGs were significantly enriched in O-glycan biosynthesis, extracellular matrix-receptor interaction, arachidonic acid metabolism, sphingolipid metabolism, as well as starch and sucrose metabolism processes. Additionally, the DEGs were found to cluster in modules pertinent to immune and energy metabolism based on weighted gene co-expression network analysis(WGCNA), hub genes like histones(H2A)and mitochondrial components(ND6)were identified. Ten DEGs were randomly selected to validate the RNA-seq results using reverse transcription-quantitative polymerase chain reaction(RT-qPCR), and the results and the expression profiles of these DEGs were of a well consistent with the high-throughput data, which confirmed the reliability of transcriptome results. 【Conclusion】 At temperatures of 32℃ and 37℃, the gills of P. clarkii are damaged. The glucose utilization rate of P. clarkii is reduced in high temperature environments, causing cellular function to be maintained through the adjustments of heat shock proteins, antioxidant capacity, and metabolism.

Key words: Procambarus clarkii, high temperature, gill, physiological metabolism, mechanism