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33
2025年5月11日 星期日
农业生物技术学报  2025, Vol. 33 Issue (5): 1106-1116    DOI: 10.3969/j.issn.1674-7968.2025.05.014
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
小黄鱼Trypsin基因克隆及其对高温胁迫和变形假单胞菌感染的响应
刘浩文1, 刘峰2,*, 张天乐2, 李倩3, 刘四芳1, 朱家杰4, 楼宝2,*, 俞晓平1
1 中国计量大学 生命科学学院,杭州 310018;
2 浙江省农业科学院 水生生物研究所/全省近岸生物种质资源保护与利用重点实验室,杭州 310021;
3 浙江海洋大学 国家海洋设施养殖工程技术研究中心,舟山 316022;
4 宁波大学 海洋学院,宁波 315832
Cloning of the Trypsin Gene in Larimichthys polyactis and Its Response to High Temperature Stress and Pseudomonas plecoglossicida Infection
LIU Hao-Wen1, LIU Feng2,*, ZHANG Tian-Le2, LI Qian3, LIU Si-Fang1, ZHU Jia-Jie4, LOU Bao2,*, YU Xiao-Ping1
1 College of Life Sciences, China Jiliang University, Hangzhou 310018, China;
2 Institute of Hydrobiology, Zhejiang Academy of Agricultural Sciences / Zhejiang Key Laboratory of Coastal Biological Germplasm Resources Conservation and Utilization, Hangzhou 310021, China;
3 National Engineering Research Center for Marine Aquaculture, Zhejiang Ocean University, Zhoushan 316022, China;
4 School of Marine Sciences, Ningbo University, Ningbo 315832, China
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摘要 夏季频发的高温以及养殖过程中变形假单胞菌(Pseudomonas plecoglossicida)引起的内脏白点病已成为制约小黄鱼(Larimichthys polyactis)养殖业发展不容忽视的问题,而参与氨基酸代谢的胰蛋白酶(trypsin)可能与小黄鱼面对高温以及变形假单胞菌感染的响应过程存在关联。为探究胰蛋白酶基因在小黄鱼应对高温胁迫和变形假单胞菌感染的响应特征,本研究克隆获得小黄鱼trypsin基因Lp_try的CDS序列共744 bp,编码247个氨基酸,含有Tryp_SPc保守结构域。组织定量分析表明Lp_try基因呈现泛组织表达特性,但是其在消化系统(肠道, 肝脏)和循环系统(心脏)中呈现特异性高表达模式。分别对32 ℃高温胁迫和变形假单胞菌感染后小黄鱼肝脏中Lp_try表达特征进行分析,发现高温胁迫导致Lp_try表达量显著增加(P<0.05),但是随着高温处理时间延长,基因表达量迅速下降;在细菌感染实验中,Lp_try表达量随着病原菌感染时间的延长呈现出降低-升高-降低的动态变化趋势,说明Lp_try在小黄鱼遭受高温胁迫和病原菌侵染过程中发挥了一定的调节作用。本研究为深入解析鱼类响应高温胁迫、病原菌感染等过程中的生理调节机制奠定了重要基础。
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刘浩文
刘峰
张天乐
李倩
刘四芳
朱家杰
楼宝
俞晓平
关键词 小黄鱼胰蛋白酶基因(try)克隆高温胁迫变形假单胞菌    
Abstract:The frequent occurrence of summer heatwaves and visceral white-nodules disease caused by Pseudomonas plecoglossicida has emerged as critical constraints on the sustainable development of small yellow croaker (Larimichthys polyactis) aquaculture. Notably, trypsin, a key enzyme in amino acid metabolism, may play a pivotal role in mediating the physiological responses of small yellow croaker to both thermal stress and bacterial infection. To investigate the response characteristics of the trypsin gene in L. polyactis under high temperature stress and P. plecoglossicida infection, the CDS sequence of the trypsin gene (Lp_try) in L. polyactis was cloned in this study. The sequence of Lp_try spaned 744 bp, encoding 247 amino acids and containing the conserved Tryp_SPc domain. Expression analysis revealed that the Lp_try gene showed pan-tissue expression characteristics, but it showed a specific high expression pattern in the digestive system (intestine, liver) and circulatory system (heart). The expression characteristics of Lp_try gene in the liver of L. polyactis subjected to 32 ℃ high temperature stress and P. plecoglossicida infection were analyzed. The results showed that high temperature stress initially induced a significant increase of Lp_try expression (P<0.05), followed by a swift decreased with prolonged exposure. Additionally, during P. plecoglossicida infection, Lp_try expression exhibited a dynamic trend of decrease-increase-decrease over time, suggesting its regulatory role in the processes of L. polyactis in response to both high temperature stress and bacterial infection. This study provides an important foundation for further understanding of the physiological regulatory mechanisms in fish in response to high temperature stress and pathogen infection.
Key wordsLarimichthys polyactis    Trypsin gene (try)    Cloning    High temperature stress    Pseudomonas plecoglossicida
收稿日期: 2024-08-21     
中图分类号: S91
基金资助:国家自然科学基金(32102765); 浙江省重点研发计划(2021C02055); 南通市科技计划重点项目(MS12022029)
通讯作者: *lengfeng0210@126.com;loubao6577@163.com   
引用本文:   
刘浩文, 刘峰, 张天乐, 李倩, 刘四芳, 朱家杰, 楼宝, 俞晓平. 小黄鱼Trypsin基因克隆及其对高温胁迫和变形假单胞菌感染的响应[J]. 农业生物技术学报, 2025, 33(5): 1106-1116.
LIU Hao-Wen, LIU Feng, ZHANG Tian-Le, LI Qian, LIU Si-Fang, ZHU Jia-Jie, LOU Bao, YU Xiao-Ping. Cloning of the Trypsin Gene in Larimichthys polyactis and Its Response to High Temperature Stress and Pseudomonas plecoglossicida Infection. 农业生物技术学报, 2025, 33(5): 1106-1116.
链接本文:  
https://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2025.05.014     或     https://journal05.magtech.org.cn/Jwk_ny/CN/Y2025/V33/I5/1106
 
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