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| 小黄鱼Gfpt1基因克隆及其对高温胁迫和变形假单胞菌感染的响应 |
| Cloning of the GFPT1 Gene in Larimichthys polyactis and Its Response to High Temperature Stress and Pseudomonas plecoglossicida Infection |
| 投稿时间：2025-03-11 修订日期：2025-04-08 |
| DOI： |
| 中文关键词: 小黄鱼，gfpt1基因，克隆，高温胁迫，变形假单胞菌 |
| 英文关键词: Larimichthys polyactis, Glutamine-fructose-6-phosphate transaminase-1 (gfpt1), Cloning, Heat stress, Pseudomonas plecoglossicida |
| 基金项目:浙江省“三农九方”科技计划项目(2025SNJF096); 国家自然科学基金项目(32102765); 浙江省重点研发计划项目(2021C02055) |
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| 中文摘要: |
| 夏季频发的高温以及养殖过程中变形假单胞菌引起的内脏白点病已成为制约小黄鱼养殖业发展不容忽视的问题。谷氨酰胺果糖-6-磷酸转氨酶-1(gfpt1)基因是信号转导和应激响应的关键调节因子。为探究gfpt1基因在小黄鱼(Larimichthys polyactis)对高温胁迫和变形假单胞菌(Pseudomonas plecoglossicida)感染的响应特征，本研究首次克隆获得小黄鱼gfpt1基因的CDS序列，序列长度2049 bp，编码682个氨基酸，含有PLN02981 super family保守结构域，同源序列比对发现其与大黄鱼相似性最高，达99.37%。荧光定量检测发现gfpt1基因在各组织中广泛表达，但表达量具有组织差异性，肝脏中的表达量显著高于其他组织。通过荧光定量PCR技术检测32℃高温胁迫和变形假单胞菌感染不同时间小黄鱼肝脏中gfpt1表达水平变化发现，高温胁迫导致gfpt1表达量显著增加，随着高温处理时间延长，基因表达量呈现先升高后降低的变化趋势，6 h时的表达量最高；变形假单胞菌感染同样导致gfpt1表达量发生显著变化，在感染后6 h的基因表达量显著低于对照组，随着感染时间的延长表达量逐渐上升，48 h时达到最高，此时显著高于对照组，随后表达量逐渐降低，至96 h时，其表达量显著低于对照组，说明gfpt1在小黄鱼响应高温胁迫和病原菌侵染过程中发挥了一定的调节作用，但是两者响应机制存在明显不同。本研究揭示了小黄鱼肝脏中gfpt1在响应高温胁迫和变形假单胞菌感染的表达变化特征，研究结果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。 |
| 英文摘要: |
| The frequent occurrence of summer heatwaves and Pseudomonas plecoglossicida-induced visceral white-nodules disease (VWND) during aquaculture practices have emerged as critical challenges hindering the sustainable development of the small yellow croaker (Larimichthys polyactis) aquaculture. The glutamine fructose-6-phosphate transaminase-1 (gftp1) gene is a critical regulator of signal transduction and stress responses To investigate the response characteristics of gfpt1 gene in L. polyactis to heat stress and P. plecoglossicida infection, this study cloned the coding sequence (CDS) of L. polyactis gfpt1 for the first time. The CDS spans 2,049 bp, encoding a 682-amino acid protein containing the conserved PLN02981 superfamily domain. Homology alignment revealed the highest sequence similarity (99.37%) with L. crocea. Expression analysis revealed that the gfpt1 gene is widely expressed across tissues but exhibits significant tissue-specific expression variation, with the highest expression observed in the liver. Quantitative real-time PCR (qRT-PCR) further demonstrated dynamic transcriptional responses of gfpt1 in L. polyactis liver to heat stress (32°C) and P. plecoglossicida challenge. Under heat stress, gfpt1 expression was significantly upregulated, peaking at 6 h, followed by a gradual decline. In contrast, P. plecoglossicida infection induced a distinct temporal pattern: gfpt1 expression decreased significantly at 6 h post-infection, followed by a significant upregulation peaking at 48 h (P<0.05), which was then followed by a significant downregulation at 96 h (P<0.05). These findings indicate that gfpt1 plays a regulatory role in both heat stress and pathogenic responses, albeit through divergent mechanisms. By integrating molecular cloning and bioinformatics analysis, this study provides pioneering insights into the molecular mechanisms underlying fish responses to environmental stressors and pathogenic invasion. These findings provide critical insights into the physiological regulatory mechanisms underlying fish responses to environmental stressors and pathogen invasion, laying a foundation for advancing aquaculture resilience research. |
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