王哲,王姿曼,郝瑞娟,李俊辉,邓岳文

• 1:广东海洋大学水产学院
• 2:广东省珍珠养殖与加工工程技术研究中心

摘要(Abstract)：

【目的】筛选(Pinctadamartensii)金黄壳色选育群体植核后免疫相关基因与代谢通路。【方法】利用转录组对植核前后马氏珠母贝的血细胞进行转录组建库及测序分析。【结果和结论】分别获得61.78×106和62.68×106个高质量转录组数据,且映射到参考基因组的平均映射率分别为70.16%和67.82%。与植核前相比,植核后分别有2 925个和3 097个基因显著上调和下调(差异倍数≥2,校正后P≤0.001),其中包括免疫相关基因凝集素、Toll样受体,热休克蛋白等。对差异基因进行GO分类分析,分为基因的分子功能、细胞组分、生物过程3个大类,并细分为11个子类别,其中结合和催化功能的基因较多。KEGG分类分析差异基因,可将基因参与的KEGG代谢通路分为6个分支,细胞过程、环境信息处理、遗传信息处理、人类疾病(仅限动物)、代谢、有机系统。KEGG富集结果显示,差异表达基因显著富集到免疫相关通路胞质DNA感应途径、白细胞跨内皮迁移等免疫通路。

关键词(KeyWords)： 马氏珠母贝;金黄壳色选育群体;转录组;免疫相关基因

Abstract:

Keywords:

基金项目(Foundation): 国家贝类产业体系专项(CARS-049);; 广东省科技厅项目(2017A030303076)

作者(Author): 王哲,王姿曼,郝瑞娟,李俊辉,邓岳文

参考文献(References)：

• [1] SOUTHGATE P C, LUCAS J S. The pearl oyster[M].Netherlands:Elsevier Science, 2008:77-102.
• [2] KISHORE P, SOUTHGATE P C. A detailed description of pearl-sac development in the black-lip pearl oyster Pinctada margaritifera(Linnaeus 1758)[J]. Aquaculture Research, 2016, 47(7):2215-2226.
• [3]尹立鹏,邓岳文,杜晓东,等.贝龄对马氏珠母贝植核贝生长、成活率和育珠性状的影响[J].中国水产科学,2012, 19(4):715-720.
• [4]林伟财,谢绍河,杜晓东,等.影响大珠母贝育珠效果的主要因素分析[J].中国农学通报, 2016, 32(32):29-33.
• [5]符韶,梁飞龙,邓岳文.影响马氏珠母贝植核育珠绩效的关键技术研究[J].水产养殖, 2016, 37(7):45-49.
• [6] BLAY C, PLANES S, KY C L. Optimal age of the donor graft tissue in relation to cultured pearl phenotypes in the mollusc, Pinctada margaritifera[J]. PLoS One, 2018,13(6):e0198505.
• [7] MCGINTY E L, EVANS B S, TAYLOR J U U, et al.Xenografts and pearl production in two pearl oyster species, P. maxima and P. margaritifera:Effect on pearl quality and a key to understanding genetic contribution[J].Aquaculture, 2010, 302(18):175-181.
• [8] SHI Y H, WANG H, ZHAN X, et al. The influence of saibo donor and host on the nacre deposits of pearls produced from Pinctada fucata martensii[J]. Journal of Shellfish Research, 2013, 32(2):271-274.
• [9]周畅,谢莉萍,张荣庆.插核手术对合浦珠母贝免疫水平的影响[J].水产学报, 2001, 25(5):419-423.
• [10]焦钰,杜晓东,师尚丽,等.珍珠囊发育中马氏珠母贝血清免疫因子的变化规律[J].广东海洋大学学报,2007, 27(4):21-24.
• [11] WEI J F, LIU B S, FAN S G, et al. Serum immune response of pearl oyster Pinctada fucata to xenografts and allografts[J]. Fish&Shellfish Immunology, 2017,62:303-310.
• [12] WANG W, LEI Q N, LIANG H Y, et al. Towards a better understanding of allograft-induced stress response in the pearl oyster Pinctada fucata martensii:Insights from iTRAQ-based comparative proteomic analysis[J]. Fish&Shellfish Immunology, 2017, 62:247-256.
• [13] WEI J F, LIU B S, FAN S G, et al. Differentially expressed immune-related genes in hemocytes of the pearl oyster Pinctada fucata against allograft identified by transcriptome analysis.[J]. Fish&Shellfish Immunology, 2017, 67:331-345.
• [14]雷倩楠.马氏珠母贝植核育珠移植耐受免疫相关基因与蛋白质鉴定及分析[D].湛江:广东海洋大学, 2016.
• [15]祁晓翔.三角帆蚌转录组和数字基因表达谱分析及EF-hand基因的表达研究[D].上海:上海海洋大学,2015.
• [16] WEI J F, FAN S G, LIU B S, et al. Transcriptome analysis of the immune reaction of the pearl oyster Pinctada fucata to xenograft from Pinctada maxima[J].Fish&Shellfish Immunology, 2017, 67:331-345.
• [17] WU Y Y, LIANG H Y, WANG Z X, et al. A novel toll-like receptor from the pearl oyster, Pinctada fucata martensii, is induced in response to stress[J].Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology, 2017, 214:19-26.
• [18] WU S Z, HUANG X D, LI Q, et al. Interleukin-17 in pearl oyster(Pinctada fucata):Molecular cloning and functional characterization[J]. Fish&Shellfish Immunology, 2013, 34(5):1050-1056.
• [19]田荣荣.马氏珠母贝胆碱能免疫调节系统的初步研究[D].湛江:广东海洋大学, 2016.
• [20]王哲,杨创业,李俊辉,等.马氏珠母贝金黄壳色选育群体与养殖群体血清免疫酶活力比较[J].中国农学通报, 2019, 35(11):160-164.
• [21] JIAO Y, YANG S, CAO Y, et al. Genome and transcriptome analyses providing insight into the immune response of pearl oysters after allograft and xenograft transplantations[J]. Fish and Shellfish Immunology, 2019, 90:109-117.
• [22]王学颖,高远镇,杜晓东,等.马氏珠母贝金黄壳色选系F3和基础群体的遗传结构比较[J].海洋通报,2012, 31(3):324-328.
• [23] DU X D, FAN G Y, JIAO Y, et al. The pearl oyster Pinctada fucata martensii genome and multi-omic analyses provide insights into biomineralization[J].GigaScience, 2017, 6(8):1–12.
• [24] WANG L L, FENG Z X, WANG X, et al. DEGseq:an R package for identifying differentially expressed genes from RNA-seq data[J]. Bioinformatics, 2010, 26(1):136-138.
• [25] FISHER W S, DINUZZO A R. Agglutination of bacteria and erythrocytes by serum from six species of marine molluscs[J]. Journal of Invertebrate Pathology, 1991,57(3):380-394.
• [26]朱家萍.马氏珠母贝血清抗菌活性蛋白的分离纯化与鉴定[D].湛江:广东海洋大学, 2015.
• [27] WANG Z L, JIAN J J, LU Y S, et al. A tandem-repeat galectin involved in innate immune response of the pearl oyster Pinctada fucata[J]. Mar Genomics, 2011, 4(3):229-236.
• [28] NI D J, SONG L S, WU L T, et al. Molecular cloning and m RNA expression of peptidoglycan recognition protein(PGRP)gene in bay scallop(Argopecten irradians, Lamarck 1819)[J]. Developmental&Comparative Immunology, 2007, 31(6), 548-558.
• [29] OLAFSEN J A, FLETCHER T C, GRANT P T.Agglutinin activity in pacific oyster(Crassostrea gigas)hemolymph following in vivo Vibrio anguillarum challenge[J]. Developmental and Comparative Immunology, 1992, 16(2/3):123-138.
• [30] TUNKIJJANUKIJ S, OLAFSEN J A. Sialic acid-binding lectin with antibacterial activity from the horse mussel:further characterization and immunolocalization[J].Developmental and Comparative Immunology, 1998,22(2):139-150.
• [31]胡钰婷,张殿昌,崔淑歌,等.合浦珠母贝C-型凝集素基因的序列特征和功能分析[J].水产学报, 2011,35(9):1327-1335.
• [32] PARK K, BANG H W, PARK J, et al. Ecotoxicological multilevel-evaluation of the effects of fenbendazole exposure to Chironomus riparius larvae[J].Chemosphere, 2009, 77(3):359-367.
• [33]王宇萍,蒋建东.热休克蛋白70的结构和功能[J].中国细胞生物学学报, 2010, 32(2):305-313.
• [34] ZHAO X X, WANG Q H, JIAO Y, et al. Identification of genes potentially related to biomineralization and immunity by transcriptome analysis of pearl sac in pearl oyster Pinctada martensii[J]. Marine Biotechnology,2012, 14(6):730-739.
• [35] GAO Q, ZHAO J M, SONG L S, et al. Molecular cloning, characterization and expression of heat shock protein 90 gene in the haemocytes of bay scallop Argopecten irradians[J]. Fish&Shellfish Immunology,2008, 24(4):379-385.
• [36]吴羽媛,郭志颖,梁海鹰,等.马氏珠母贝TLR6基因的克隆、序列分析与表达[J].水产学报, 2017, 41(11):1687-1697.
• [37] GERDOL M, GOMEZ-CHIARRI M, CASTILLO M G,et al. Immunity in molluscs:Recognition and effector mechanisms, with a focus on Bivalvia[M] Advances in Comparative Immunology. 2018, 8:225-341.
• [38] LI R J, ZHANG R, ZHANG L, et al. Characterizations and expression analyses of NF-κB and Rel genes in the Yesso scallop(Patinopecten yessoensis)suggest specific response patterns against Gram-negative infection in bivalves[J]. Fish&Shellfish Immunology, 2015, 44(2):611-621.
• [39] MYLèNE T, GERDOL M, ROSANI U, et al. Toll-like receptors and MyD88 adaptors in Mytilus:Complete CDS and gene expression levels[J]. Developmental&Comparative Immunology, 2013, 40(2):158-166.
• [40] TOUBIANA, M, ROSANI U, GIAMBELLUCA S, et al.Toll signal transduction pathway in bivalves:Complete CDS of intermediate elements and related gene transcription levels in hemocytes of immune stimulated Mytilus galloprovincialis[J]. Developmental&Comparative Immunology, 2014, 45(2):300-312.