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| Transcriptome Analysis of Hepatopancreas Tissue with Non-O1 Vibrio cholerae Infection in Giant Freshwater Prawn Macrobrachium rosenbergii |
| ZHANG Zirui, ZHU Xinhai, ZHANG Xiaojun, GAO Xiaojian, SUN Yongjun, WEI Wansheng, JIANG Qun |
| College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China |
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Abstract In 2018, outbreaks of mass mortalities occurred in giant freshwater prawn Macrobrachium rosenbergii farms in Gaoyou city, Jiangsu Province of China, causing significant economic damage, and Non-O1 Vibrio cholerae is recognized to be the etiological agent of the enzootic. To investigate the immune response, giant freshwater prawn [weight(7.22±0.46)g] was infected withNon-O1 V. cholerae (1×106 cfu/mL) for 24 h. Hepatopancreas tissue with and without non-O1 V. cholerae infection was sampled and then comparative transcriptome analysis was conducted using Illumina Hiseq 2000. After de novo assembly, 85794 high quality unigenes were obtained with an average length of 1208 bp (N50=2804 bp). Searching against the public databases, including GO, COG, KEGG, NR, SWISS and NT, 29731 unigenes were annotated. Differential expression analysis showed that 3236 significantly differently expressed genes (DEGs) were identified with 1745 up-regulated genes and 1419 down-regulated genes. According to the information of gene function annotation, 16 immune-related genes were obtained. The expression level of nine well-known immune-related genes, including LEC 3 and Serpin1, significantly increased. While other seven genes, including HMC and SCARB, showed significantly decreased expression level. The dominant enriched GO terms of DEGs were involved in cellular process, cell and binding. Three GO terms, including immune system process, response to stimulus, and antioxidant activity, involved in immune response, were enriched. This result demonstrated that non-O1 V. cholerae infection stimulated and activate the antioxidant system and further triggers non-self-recognition molecules and immune response. KEGG analysis revealed that DEGs were enriched into 121 pathways including several immune-related pathways, such as phagosome, ECM-receptor interaction, Vibrio cholerae infection, leukocyte transendothelial migration, NOD-like receptor signaling pathway and so on. To verify the accuracy of transcriptome sequencing, eight DEGs were selected to conduct quantitative real-time PCR. Results of qRT-PCR were consistent with RNA-seq, which indicated that the differential gene expression results based on the transcriptome sequencing were credible. Data from this study provided part of basic information about gene expression changes of hepatopancreas after non-O1 V. cholerae infection, which might be helpful for further studying the immune response of giant freshwater prawn against non-O1 V. cholerae infection.
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Received: 24 September 2021
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[1] 苗鹏飞,杨映,谭淑雯,等.罗氏沼虾致病性溶藻弧菌的鉴定及药敏分析[J].水产科学,2018,37(3):384-388.
[2] 唐琼英,夏正龙,蔡缪荧,等.罗氏沼虾养殖群体表型性状间的相关性及类群差异分析[J].中国水产科学,2019,26(6):1075-1085.
[3] 童桂香,黎小正,卢小花,等.罗氏沼虾诺达病毒套式RT-PCR检测方法的建立及初步应用[J].上海海洋大学学报,2012,21(1):54-59.
[4] 钱冬,石正丽,曹铮,等.罗氏沼虾苗种肌肉白浊病诺达病毒的分离和特性研究[J].中国水产科学,2003,10(6):457-461.
[5] 潘晓艺,刘杜鹃,沈锦玉,等.罗氏沼虾野田村病毒和双顺反子病毒双重RT-PCR检测方法与序列分析[J].上海海洋大学学报,2012,21(6):996-1002.
[6] 陶保华,石和荣,黄俊文,等.假单胞菌引起罗氏沼虾黄鳃、黑鳃病的研究[J].中山大学学报(自然科学版),2000,39(S1):255-259.
[7] 郭莹,周淼,李艳和,等.罗氏沼虾豚鼠气单胞菌的分离鉴定[J].中国农学通报,2020,36(33):147-153.
[8] 陈雪峰,杨国梁,高强,等.罗氏沼虾(Macrobrachium rosenbergii)幼体新暴发病病原阴沟肠杆菌的分离鉴定[J].海洋与湖沼,2015,46(6):1467-1477.
[9] 朱其建,戴习林,邹卫丽,等.罗氏沼虾抗病选育群体的抗病性能及其遗传多样性分析[J].水产学报,2013,37(10):1468-1478.
[10] 郭莹,郭青松,周淼,等.罗氏沼虾维氏气单胞菌的分离鉴定[J].中国农学通报,2020,36(36):137-143.
[11] GAO X J,MIAO Z,LI X X,et al.Pathogenicity of non-O1/O139 Vibrio cholerae and its induced immune response in Macrobrachium rosenbergii[J].Fish & Shellfish Immunology,2019,92:300-307.
[12] HARRIS J B,LAROCQUE R C,QADRI F,et al.Cholera[J].Lancet,2012,379(9835):2466-2476.
[13] LI X X,YANG H,GAO X J,et al.The pathogenicity characterization of non-O1 Vibrio cholerae and its activation on immune system in freshwater shrimp Macrobrachium nipponense[J].Fish & Shellfish Immunology,2019,87:507-514.
[14] GAO X J, CHEN N, ZHANG Y, et al. Non-O1 Vibrio cholerae pathogen from Cyprinus carpio and control with anti-non-O1 V. cholerae egg yolk powder (IgY)[J]. Aquaculture, 2017, 479: 69-74.
[15] 祝令伟,景洁,梁冰,等.野生水鸟感染霍乱弧菌和沙门菌等病原菌的分离和鉴定[J].中国人兽共患病学报,2019,35(3):212-215.
[16] HALDAR S,CHATTERJEE S,ASAKURA M,et al.Isolation of Vibrio parahaemolyticus and Vibrio cholerae (Non-O1 and O139) from moribund shrimp (Penaeus monodon) and experimental challenge study against post larvae and juveniles[J].Annals of Microbiology,2007,57(1):55-60.
[17] ZHANG X J.Identification of Vibrio cholerae as a causative bacterium for an ulcer disease of cultured loach Misgurnus anguillicaudatus in China[J].African Journal of Microbiology Research,2012,6(9):2060-2070.
[18] 秦蕾,徐静,张晓君.异育银鲫非O1/非O139群霍乱弧菌的分离及鉴定[J].中国预防兽医学报,2013,35(2):130-133.
[19] SOOKRUKSAWONG S,SUN F Y,LIU Z J,et al.RNA-Seq analysis reveals genes associated with resistance to Taura syndrome virus (TSV) in the Pacific white shrimp Litopenaeus vannamei[J].Developmental and Comparative Immunology,2013,41(4):523-533.
[20] 苗珍,李席席,高晓建,等.中华绒螯蟹病原弗氏柠檬酸杆菌的鉴定及其毒力基因检测[J].畜牧与兽医,2020,52(10):85-90.
[21] 张天天,吴征卓,陈国权,等.1株鸭源非O1/O139群霍乱弧菌的分离鉴定与致病性分析[J].中国畜牧兽医,2021,48(4):1457-1465.
[22] 王苗,张雪佳,杜万年,等.1株草龟维氏气单胞菌的分离鉴定及药敏谱分析[J].畜牧兽医学报,2020,51(10):2528-2535.
[23] SALZET M.Vertebrate innate immunity resembles a mosaic of invertebrate immune responses[J].Trends in Immunology,2001,22(6):285-288.
[24] RUNSAENG P,PUENGYAM P,UTARABHAND P.A mannose-specific C-type lectin from Fenneropenaeus merguiensis exhibited antimicrobial activity to mediate shrimp innate immunity[J].Molecular Immunology,2017,92:87-98.
[25] SUN Y D,FU L D,JIA Y P,et al.A hepatopancreas-specific C-type lectin from the Chinese shrimp Fenneropenaeus chinensis exhibits antimicrobial activity[J].Molecular Immunology,2008,45(2):348-361.
[26] LI M,LI C Z,MA C X,et al.Identification of a C-type lectin with antiviral and antibacterial activity from Pacific white shrimp Litopenaeus vannamei[J].Developmental and Comparative Immunology,2014,46(2):231-240.
[27] 刘勇杰.凡纳滨对虾免疫相关基因QM和SERPIN的功能研究[D].杨凌:西北农林科技大学,2014.
[28] 陶梦圆,李长平,伦镜盛,等.源于凡纳滨对虾血蓝蛋白的化学合成肽段的免疫调控活性[J].中国水产科学,2019,26(5):844-851.
[29] ALINEJAD T,BIN K Q,VEJAYAN J,et al.Proteomic analysis of differentially expressed protein in hemocytes of wild giant freshwater prawn Macrobrachium rosenbergii infected with infectious hypodermal and hematopoietic necrosis virus (IHHNV)[J].Meta Gene,2015,5:55-67.
[30] 高晓建.罗氏沼虾病原阴沟肠杆菌致病性及rpoS基因在其生存和毒力中的功能研究[D].扬州:扬州大学,2021:27-48.
[31] 吴羽媛,郭志颖,梁海鹰,等.马氏珠母贝(Pinctada fucata martensii)新型清道夫受体基因克隆与表达分析[J].基因组学与应用生物学,2018,37(2):733-740.
[32] 陈永华,蒋建新,谢国旗,等.内毒素血症时小鼠肺泡巨噬细胞CD14和清道夫受体的表达[J].第三军医大学学报,2000,22(7):615-618.
[33] ZHOU Y F,ZHU X H,TANG H Y,et al.Immune related gene expression analysis of Macrobrachium nipponense in different hours post-infection by non-O1 Vibrio cholerae[J].Aquaculture Reports,2021,19:100571.
[34] RAO R,ZHU Y B,ALINEJAD T,et al.RNA-seq analysis of Macrobrachium rosenbergii hepatopancreas in response to Vibrio parahaemolyticus infection[J].Gut Pathogens,2015,7:6. |
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