云龙石斑鱼（Epinephelus moara♀和Epinephelus lanceolatus♂）幼虫细胞系的建立、鉴定及其对感染性脾肾坏死病毒感染的反应

IF 3.9 1区农林科学 Q1 FISHERIES

Aquaculture Pub Date : 2025-04-07 DOI:10.1016/j.aquaculture.2025.742476

Chenfei Guo , Zhihong Gong , Fei Fang , Qi Zhang , Na Wang , Yuqin Su , Lei Wang , Zhenxia Sha , Songlin Chen

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引用次数: 0

摘要

云龙石斑鱼是由石斑鱼母与石斑鱼公杂交而成的新杂交种。这种杂交种表现出从两个亲本种遗传而来的理想性状的组合。本研究建立了云龙石斑鱼幼虫细胞系，命名为Glar （grouper幼虫）。细胞在24℃含15%血清的L-15培养基中培养，培养80代以上。染色体核型分析显示染色体数目为2n = 48，转录组分析表明该细胞系主要由成纤维细胞组成。在液氮中低温保存6个月后，细胞存活率保持在70%以上。细胞可以转染，转染cy3标记的scramble siRNA和pEGFP-N1质粒后均观察到荧光。此外，两种虹膜病毒，斑点刀颌虹膜病毒（SKIV-SD）和斑点鲈鱼虹膜病毒（SBIV-V12）感染后，玻璃细胞表现出明显的细胞病变效应（CPE），并在透射电镜下观察到病毒颗粒。转录组分析显示，感染SBIV-V12 5天后，差异表达基因主要富集于与细胞复制、细胞周期、DNA修复、p53信号通路和Fanconi贫血通路相关的通路，其中p53通路21个基因中有20个基因表达上调。p53信号通路在抗病毒免疫中起关键作用。因此，该细胞系可为虹膜病毒的体外防治研究、研究云龙石斑鱼的发病机制和潜在的治疗策略提供有价值的工具。

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Establishment and characterization of a cell line from Yunlong grouper (Epinephelus moara ♀ and Epinephelus lanceolatus ♂) larva and its response to infectious spleen and kidney necrosis virus infection

The Yunlong grouper is a novel hybrid species resulting from the crossbreeding of Epinephelus moara ♀ and Epinephelus lanceolatus ♂. This hybrid species exhibits a combination of desirable traits inherited from both parent species. This study established the first Yunlong grouper cell line from larvae, named Glar (Grouper larvae).

The cells were cultured in L-15 medium containing 15 % serum in 24 °C and have culture more than 80 passages. Chromosome karyotype analysis showed that the chromosome number was 2n = 48, and transcriptome analysis indicated that the cell line is mainly composed of fibroblast. After cryopreservation in liquid nitrogen for 6 months, cell viability remained above 70 %. The cells can be transfected, as fluorescence was observed after transfection with both Cy3-labeled scramble siRNA and the pEGFP-N1 plasmid. Furthermore, after infection with two iridoviruses, spotted knifejaw iridovirus (SKIV-SD) and spotted seabass iridovirus (SBIV-V12), Glar cells showed significant cytopathic effect (CPE), and viral particles were observed using transmission electron microscopy. Transcriptome analysis revealed that five days post-infection with SBIV-V12, differentially expressed genes were primarily enriched in pathways related to cell replication, the cell cycle, DNA repair, the p53 signaling pathway, and the Fanconi anaemia pathway, with 20 of 21 genes in the p53 pathway showing upregulation. The p53 signaling pathway plays a crucial role in antiviral immunity. Therefore, this cell line can serve as a valuable tool for in vitro research on the prevention and control of iridovirus, as well as for studying the disease mechanisms and potential therapeutic strategies for the Yunlong grouper.

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来源期刊

Aquaculture 农林科学-海洋与淡水生物学

CiteScore

8.60

自引率

17.80%

发文量

1246

审稿时长

56 days

期刊介绍： Aquaculture is an international journal for the exploration, improvement and management of all freshwater and marine food resources. It publishes novel and innovative research of world-wide interest on farming of aquatic organisms, which includes finfish, mollusks, crustaceans and aquatic plants for human consumption. Research on ornamentals is not a focus of the Journal. Aquaculture only publishes papers with a clear relevance to improving aquaculture practices or a potential application.