大口黑鲈头肾抗大口黑鲈虹彩病毒(LMBV)应答的转录组学分析

doi:10.11924/j.issn.1000-6850.casb2024-0440

中国农学通报 ›› 2024, Vol. 40 ›› Issue (35): 148-155.doi: 10.11924/j.issn.1000-6850.casb2024-0440

大口黑鲈头肾抗大口黑鲈虹彩病毒(LMBV)应答的转录组学分析

张晶晶(), 陈斌, 卓玉琛, 翁可佳, 林而舒, 陈宇舒, 樊海平()

福建省淡水水产研究所，福州 350002

收稿日期:2024-07-05 修回日期:2024-09-09 出版日期:2024-12-15 发布日期:2024-12-12
通讯作者:
樊海平，男，1967年出生，江苏武进人，研究员，硕士，研究方向：水产养殖病害研究。通信地址：350002福建省福州市鼓楼区西洪路555号福建省淡水水产研究，Tel：0591-87640080，E-mail：fanhaiping16@163.com。
作者简介:
张晶晶，女，1992年出生，福建莆田人，农艺师，硕士，研究方向：水产养殖病害研究。通信地址：350002福建省福州市鼓楼区西洪路555号福建省淡水水产研究所，Tel：0591-83732007，E-mail：849173247@qq.com。
基金资助:
2023年度福建省海洋服务与渔业高质量发展专项资金项目“大口黑鲈蛙虹彩病毒病的研究及免疫制剂的初步研制”(FJHY-YYKJ-2023-2-5); 2023年度福建省海洋服务与渔业高质量发展专项资金项目“大口黑鲈‘优鲈3号’工厂化育苗技术研究”(FJHY-YYKJ-2023-2-3)

Transcriptome Analysis of Head Kidney Anti-LMBV Responses in Largemouth Bass (Micropterus salmoides)

ZHANG Jingjing(), CHEN Bin, ZHUO Yuchen, WENG Kejia, LIN Ershu, CHEN Yushu, FAN Haiping()

Freshwater Fisheries Research Institute of Fujian province, Fuzhou 350002

Received:2024-07-05 Revised:2024-09-09 Published:2024-12-15 Online:2024-12-12

摘要/Abstract

摘要：

为了分析大口黑鲈(Largemouth bass，Micropterus salmoides)抗大口黑鲈蛙虹彩病毒(Largemouth bass ranavirus, LMBV)转录组图谱，揭示大口黑鲈响应LMBV感染的免疫应答机制。利用Illumina NovaSeq6000测序平台，对感染LMBV 72 h后的大口黑鲈头肾组织进行转录组测序分析。研究共获得5953个差异表达基因(Differentially expressed genes, DEGs)，其中1704个上调表达基因，4249个下调表达基因。通过GO功能注释发现，差异表达基因与单有机体过程、代谢过程、膜、细胞、结合、催化活性等功能相关。KEGG富集分析结果显示，差异表达基因主要富集于糖酵解、初级胆汁酸生物合成等代谢途径。与免疫应答相关的信号通路4条，包括细胞因子与细胞因子受体相互作用、细胞外基质受体互作、吞噬体和免疫球蛋白A产生的肠道免疫网络。免疫应答相关DEGs提高的为cxcr4、il10、mrc、ncf4、itgb2、il27、ccl25等；下降的DEGs为cxcl12、tgfb3、il20ra、col4a5、itgb1等。PPI蛋白互作网络分析显示itgb1、itga8和itgb6为免疫相关DEGs的核心基因。本研究分析了大口黑鲈感染LMBV的转录组图谱，为大口黑鲈抗LMBV免疫的分子机制和疾病防控研究提供理论基础。

关键词: 大口黑鲈, LMBV, 转录组, 免疫应答

Abstract:

The transcription profile of Micropterus salmoides against LMBV infection was analyzed to reveal the immune response mechanism of Micropterus salmoides in response to LMBV infection. The head kidney tissues of largemouth bass infected with LMBV 72h were analyzed by transcriptome sequencing using Illumina NovaSeq 6000 sequencing platform. A total of 5953 differentially expressed genes (DEGs) were obtained, of which 1704 were up-regulated and 4249 were down-regulated. Through GO function annotation, DEGs were mainly related to single-organism process, metabolic process, membrane, cell, binding, catalytic activity, etc. The KEGG enrichment analysis showed that DEGs were mainly enriched in metabolic pathways such as glycolysis, primary bile acid biosynthesis. There were 4 immune response-related signaling pathways enriched including cytokine-cytokine receptor interaction, ECM-receptor interaction, phagosome and intestinal immune network for IgA production. The immune-related increased DEGs were cxcr4, il10, mrc, ncf4, itgb2, il27, ccl25, etc. while the decreased degs were cxcl12, tgfb3, il20ra, col4a5, itgb1, etc. PPI analysis showed that itgb1, itga8 and itgb6 were the hub genes of immune-related DEGs. This study analyzed the transcription profile of largemouth bass infected with LMBV to provide a theoretical basis for the molecular mechanism and disease prevention of largemouth bass immunity against LMBV.

Key words: Micropterus salmoides, LMBV, transcriptome, immune response

张晶晶, 陈斌, 卓玉琛, 翁可佳, 林而舒, 陈宇舒, 樊海平. 大口黑鲈头肾抗大口黑鲈虹彩病毒(LMBV)应答的转录组学分析[J]. 中国农学通报, 2024, 40(35): 148-155.

ZHANG Jingjing, CHEN Bin, ZHUO Yuchen, WENG Kejia, LIN Ershu, CHEN Yushu, FAN Haiping. Transcriptome Analysis of Head Kidney Anti-LMBV Responses in Largemouth Bass (Micropterus salmoides)[J]. Chinese Agricultural Science Bulletin, 2024, 40(35): 148-155.

图/表 9

参考文献 22

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基因名称	引物(5′—3′)
itga6	GCTAAACGGAAACGCAGAC
itga6	ACCCTCCTTCAGGAAATTCACCA
ccr9	TCCATCAGCCCTTCACCC
ccr9	CCAGCCATTCAACGCCTC
cxcr2	CCTGGAAATCTGGTCGTG
cxcr2	CCTGCGAGTCTCCATAGC
ctss	TCAAAGGAAAGGCTGGCTAC
ctss	TGTCCTCCGTCTGTGCC
itgb2	GTGTCTGTGGCTCTTATCG
itgb2	CCATTTGGCTTTCTTCTT
ncf1	TGTCTCAGTACCACCTGGGTCAC
ncf1	CTGGCTTTCCGTCAGTTTGCTCT
β-actin	CCACCACAGCCGAGAGGGAA
β-actin	TCATGGTGGATGGGGCCAGG

基因名称	引物(5′—3′)
itga6	GCTAAACGGAAACGCAGAC
itga6	ACCCTCCTTCAGGAAATTCACCA
ccr9	TCCATCAGCCCTTCACCC
ccr9	CCAGCCATTCAACGCCTC
cxcr2	CCTGGAAATCTGGTCGTG
cxcr2	CCTGCGAGTCTCCATAGC
ctss	TCAAAGGAAAGGCTGGCTAC
ctss	TGTCCTCCGTCTGTGCC
itgb2	GTGTCTGTGGCTCTTATCG
itgb2	CCATTTGGCTTTCTTCTT
ncf1	TGTCTCAGTACCACCTGGGTCAC
ncf1	CTGGCTTTCCGTCAGTTTGCTCT
β-actin	CCACCACAGCCGAGAGGGAA
β-actin	TCATGGTGGATGGGGCCAGG

样本名	原始序列/条	有效序列/条	有效碱基/Gb	错误率/%	Q20/%	Q30/%	GC含量/%
EK1	40716768	40265536	6.04G	0.02	98.09	94.67	48.28
EK2	40235984	39754866	5.96G	0.02	98.09	94.61	48.93
EK3	41964132	41441194	6.22G	0.02	98.19	94.86	49.05
BK1	51406336	50946762	7.64G	0.03	96.68	91.97	47.84
BK2	41622308	41160564	6.17G	0.02	98.13	94.78	47.53
BK3	47646868	47093288	7.06G	0.02	98.24	94.89	49.46

样本名	原始序列/条	有效序列/条	有效碱基/Gb	错误率/%	Q20/%	Q30/%	GC含量/%
EK1	40716768	40265536	6.04G	0.02	98.09	94.67	48.28
EK2	40235984	39754866	5.96G	0.02	98.09	94.61	48.93
EK3	41964132	41441194	6.22G	0.02	98.19	94.86	49.05
BK1	51406336	50946762	7.64G	0.03	96.68	91.97	47.84
BK2	41622308	41160564	6.17G	0.02	98.13	94.78	47.53
BK3	47646868	47093288	7.06G	0.02	98.24	94.89	49.46

大口黑鲈头肾抗大口黑鲈虹彩病毒(LMBV)应答的转录组学分析

Transcriptome Analysis of Head Kidney Anti-LMBV Responses in Largemouth Bass (Micropterus salmoides)

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