| 摘要: |
| 水产养殖环境是影响鱼类福利的关键因素。其中, 光谱环境显著影响养殖鱼类的生长发育、生理状态和行为特征。近年来, 随着工厂化养殖模式的快速发展, 人工光源获得广泛应用, 关于养殖鱼类光环境调控的研究日益增多, 光谱环境优化已成为推动精准化、集约化养殖发展的重要环节。然而, 在相关基因表达分析中, 内参基因的选择仍缺乏充分依据。本研究以大菱鲆幼鱼为研究对象, 利用实时荧光定量PCR(qRT-PCR)技术, 结合geNorm、Normfinder和Bestkeeper三种算法, 系统分析了5种光谱条件下肝脏和肌肉组织中7个候选内参基因(18S、actb、rpl17、tuba、ef1a、b2m和rpsd)的表达稳定性。结果表明, 不同光谱对肝脏和肌肉中内参基因的表达水平具有显著影响。内参基因的稳定性存在组织特异性: 在肝脏中, 最佳双内参基因组合为actb和b2m, 最佳单内参基因为ef1a或b2m; 在肌肉中, 最佳双内参基因组合为rpl17和rpsd, 最佳单内参基因为tuba。本研究筛选获得的稳定内参基因, 为后续大菱鲆养殖系统光谱环境优化研究中的基因表达分析提供了可靠校准依据。 |
| 关键词: 大菱鲆 光谱 内参基因 实时荧光定量 表达稳定性 |
| DOI：10.11759/hykx20250827003 |
| 分类号:Q955;S965 |
| 基金项目:国家自然科学基金青年基金(32402989); 国家重点研发计划(2024YFD2400101); 国家自然科学基金(32273104); 山东省博士后创新项目(SDCX-ZG-202302005); 青岛市博士后资助项目(QDBSH20230202089);设施渔业教育部重点实验室(大连海洋大学)资助项目(116023) |
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| Expression levels and stability of reference genes in the muscles and livers of juvenile turbots (Scophthalmus maximus) under different light spectra |
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Zhou Jiale1, Li Xian1, Song Zongcheng2, Wu Lele1
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1.Fisheries College, Ocean University of China, Qingdao 266003, China;2.Weihai Shenghang Aquatic Product Science and Technology Co., Ltd., Weihai 264200, China
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| Abstract: |
| Optimization of the light spectrum in aquaculture systems is an important component of fish welfare. In recent years, the research on optimizing such a spectral environment has become crucial. In this study, the expression levels and stability of seven candidate internal reference genes—18S, actb, rpl17, tuba, ef1a, b2m, and rpsd—in livers and muscles were analyzed by qRT-PCR as well as geNorm, Normfinder, and Bestkeeper software. The results showed that the spectrum considerably impacts the expression levels of reference genes in livers and muscles. The expression stability of internal reference genes was tissue-specific. In livers, the best combination of two internal reference genes was actb and b2m, and the best reference gene combination is ef1a or b2m. In muscles, the best combination of two internal reference genes was rpl17 and rpsd, and the best internal reference gene was tuba. The results provide a calibration basis for subsequent studies on spectrum optimization for juvenile turbots in aquaculture. |
| Key words: Scophthalmus maximus light spectrum internal control gene real-time fluorescence quantitative expression stability |
