菲牛蛭肠道细菌的群落结构及功能
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摘要
为了揭示肠道细菌对菲牛蛭(Hirudinaria manillensis)消化、代谢等生理活动的影响,采用二代测序技术测定了三个菲牛蛭个体肠道细菌的群落结构并推断了其功能。结果共鉴定出细菌7门,16纲,24目,29科,36属,40个操作分类单元(operation taxonomic units,OTUs);其中优势属分别为色杆菌属(Chromobacterium)、甲基菌属(Methylovorus)和Jonquetella。采用PICRUSt软件对菲牛蛭肠道细菌进行功能预测,结果显示其主要功能依次是膜转运、碳水化合物代谢、氨基酸代谢、细胞迁移、能量代谢、基因复制和修复、辅酶因子和维生素代谢、外源物质降解、细胞进程和信号转导等。
To reveal the effects of gut bacteria on the digestion,metabolism and other physiological functions of Hirudinaria manillensis and to provide the theoretical foundation for the aquaculture and medicinal purposes on leeches,this paper demonstrates the community structure and functions of gut bacteria of Hirudinaria manillensis using the next-generation sequencing technique.Forty OTUs were identified,which belong to 36 genera,29 families,24 orders,16 classes and 7 phylums.Among the three samples,the dominant genera were Chromobacterium,Methylovorus and Jonquetella.The functions of these gut bacteria were further predicted using the software PICRUSt.The results showed that the dominent functional category was the membrane transport,followed by the carbohydrate metabolism,amino acid metabolism,cell motility,energy metabolism,DNA replication and repair,metabolism of cofactors and vitamins,xenobiotics biodegradation metabolism,cellular processes and signaling.
To reveal the effects of gut bacteria on the digestion,metabolism and other physiological functions of Hirudinaria manillensis and to provide the theoretical foundation for the aquaculture and medicinal purposes on leeches,this paper demonstrates the community structure and functions of gut bacteria of Hirudinaria manillensis using the next-generation sequencing technique.Forty OTUs were identified,which belong to 36 genera,29 families,24 orders,16 classes and 7 phylums.Among the three samples,the dominant genera were Chromobacterium,Methylovorus and Jonquetella.The functions of these gut bacteria were further predicted using the software PICRUSt.The results showed that the dominent functional category was the membrane transport,followed by the carbohydrate metabolism,amino acid metabolism,cell motility,energy metabolism,DNA replication and repair,metabolism of cofactors and vitamins,xenobiotics biodegradation metabolism,cellular processes and signaling.
引文
[1]王珊珊,王佳堃,刘建新.肠道微生物对宿主免疫系统的调节及其可能机制[J].动物营养学报,2015,27(2)375-382.
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[10]张广美,姜梅.水蛭抗肿瘤作用探讨[J].中华中医药学刊,2009,27(11):2257-2258.
[11]李克明,张国,武继彪.水蛭的药理研究概况[J].中医研究,2007,20(2):62-64.
[12]刘君,杨松松,栾淑华.对不同食性的水蛭药效价值的探讨[J].中医药学刊,2005,23(4):705-706.
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[14]张彬,龚元,于翔,等.体重、年龄和应激刺激对菲牛蛭繁殖性能的影响[J].四川动物,2012,31(3):397-401.
[15]李文廖,福龙,殷晓杰,等.七种水蛭抗血小板聚集与抗凝血研究[J].中药药理与临床,1997,13(5):33-35.
[16]魏恒,李敏,段红宇,等.不同试剂处理对菲牛蛭冻干粉抗凝活性的影响研究[J].成都中医药大学学报,2015,38(1):37-40.
[17]GRAF J,KIKUCHI Y,RIO R V M.Leeches and their microbiota:naturally simple symbiosis models[J].Trends in Microbiology,2006,14(8):365-371.
[18]MALTZ M A,BOMAR L,LAPIERRE P,et al.Metagenomic analysis of the medicinal leech gut microbiota[J].Frontiers in Microbiology,2014,5(151):1-12.
[19]LAUFER A S,SIDDALL M E,GRAF J.Characterization of the digestivetract microbiota of Hirudo orientalis,a European medicinal leech[J].Applied and Environmental Microbiology,2008,74(19):6151-6154.
[20]CAPORASO J G,LAUBER C L,WALTERS W A,et al.Global patterns of 16srRNA diversity at a depth of millions of sequences per sample[J].Proceedings of the National Academy of Sciences of the United States of America,2011,108:4516-4522.
[21]EDGAR R C.UPARSE:highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods,2013,10(10):996-998.
[22]LANGILLE,M G I,ZANEVELD J,CAPORASO J G,et al.Predictive functional profiling of microbial communities using 16SrRNA marker gene sequences[J].Nature.Biotechnology,2013,31:814-823.
[23]MARTIN P A W,GUNDERSENRINDAL D,BLACKBURN M,et al.Chromobacterium subtsugae sp.nov.a betaproteobacterium toxic to Colorado potato beetle and other insect pests[J].International Journal of Systematic&Evolutionary Microbiology,2007,57(5):993-999.
[24]YAMAMOTO S,MORIHARA Y,WAKAYAMA M,et al.Theanine production by coupled fermentation with energy transfer usingγ-glutamylmethylamide synthetase of Methylovorus mays No.9[J].Bioscience Biotechnology&Biochemistry,2008,72(5):1206-1211.
[25]PHILIP H,HOOPER S D,KYRPIDES N C.Focus:synergistetes[J].Environmental Microbiology,2009,11(6):1327-1329.
[26]WORTHEN P L,GODE C J,GRAF J.Culture-independent characterization of the digestive-tract microbiota of the medicinal leech reveals a tripartite symbiosis[J].Applied&Environmental Microbiology,2006,72(7):4775-81.
[27]杨预展.长江中下流域草食性雁食性及肠道微生物研究[D].合肥:中国科学技术大学生命科学学院,2016.
[28]WAITE D W,TAYLOR M W.Characterizing the avian gut microbiota:membership,driving influences,and potential function[J].Frontiers in Microbiology,2014,5(4):1-12.
[29]ZHANG B,LIN Q,LIN J,et al.Effects of broodstock density and diet on reproduction and juvenile culture of the Leech,Hirudinaria manillensis,Lesson,1842[J].Aquaculture,2008,276(1/4):198-204.
[2]王春晖,张华玲,张祺玲,等.超微七味白术散对肠道厌氧微生物代谢多样性的调控作用[J].生态学报,2015,35(14):4843-4851.
[3]周雪雁,刘翊中,陈轶霞,等.动物肠道菌群结构分析方法进展[J].微生物学杂志,2013,33(5):81-86.
[4]杨丽平,常会会,李杰,等.基于DNA复合条形码技术的蝗虫肠道共生真菌多样性研究[J].生态学报,2017,37(20):1-9.
[5]KOSTROMINA M A,ESIPOV R S,MIROSHNIKOV A I.Biotechnological production of recombinant analogs of hirudin-1from Hirudo medicinalis[J].Bioorganicheskaia Khimiia,2012,38(2):166-176.
[6]MING S,SARKAR I N,SIDDALL M E.Salivary transcriptome of the north american medicinal leech,Macrobdella decora[J].Journal of Parasitology,2010,96(6):1211-1221.
[7]DUNWIDDIE C,THORNBERRY N A,BULL H G,et al.Antistasin,a leech-derived inhibitor of factor Xa.Kinetic analysis of enzyme inhibition and identification of the reactive site[J].Journal of Biological Chemistry,1989,264(28):16694-16699.
[8]MARKWARDT F.The development of hirudin as an antithrombotic drug[J].Thrombosis Research,1994,74(1):1-23.
[9]张英华,杨白玉,王永生,等.动物药水蛭的药理、临床研究[J].长春中医学院学报,1994,10(40):52-52.
[10]张广美,姜梅.水蛭抗肿瘤作用探讨[J].中华中医药学刊,2009,27(11):2257-2258.
[11]李克明,张国,武继彪.水蛭的药理研究概况[J].中医研究,2007,20(2):62-64.
[12]刘君,杨松松,栾淑华.对不同食性的水蛭药效价值的探讨[J].中医药学刊,2005,23(4):705-706.
[13]杨潼.中国动物志-环节动物门-蛭纲[M].北京:科学出版社,1996.
[14]张彬,龚元,于翔,等.体重、年龄和应激刺激对菲牛蛭繁殖性能的影响[J].四川动物,2012,31(3):397-401.
[15]李文廖,福龙,殷晓杰,等.七种水蛭抗血小板聚集与抗凝血研究[J].中药药理与临床,1997,13(5):33-35.
[16]魏恒,李敏,段红宇,等.不同试剂处理对菲牛蛭冻干粉抗凝活性的影响研究[J].成都中医药大学学报,2015,38(1):37-40.
[17]GRAF J,KIKUCHI Y,RIO R V M.Leeches and their microbiota:naturally simple symbiosis models[J].Trends in Microbiology,2006,14(8):365-371.
[18]MALTZ M A,BOMAR L,LAPIERRE P,et al.Metagenomic analysis of the medicinal leech gut microbiota[J].Frontiers in Microbiology,2014,5(151):1-12.
[19]LAUFER A S,SIDDALL M E,GRAF J.Characterization of the digestivetract microbiota of Hirudo orientalis,a European medicinal leech[J].Applied and Environmental Microbiology,2008,74(19):6151-6154.
[20]CAPORASO J G,LAUBER C L,WALTERS W A,et al.Global patterns of 16srRNA diversity at a depth of millions of sequences per sample[J].Proceedings of the National Academy of Sciences of the United States of America,2011,108:4516-4522.
[21]EDGAR R C.UPARSE:highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods,2013,10(10):996-998.
[22]LANGILLE,M G I,ZANEVELD J,CAPORASO J G,et al.Predictive functional profiling of microbial communities using 16SrRNA marker gene sequences[J].Nature.Biotechnology,2013,31:814-823.
[23]MARTIN P A W,GUNDERSENRINDAL D,BLACKBURN M,et al.Chromobacterium subtsugae sp.nov.a betaproteobacterium toxic to Colorado potato beetle and other insect pests[J].International Journal of Systematic&Evolutionary Microbiology,2007,57(5):993-999.
[24]YAMAMOTO S,MORIHARA Y,WAKAYAMA M,et al.Theanine production by coupled fermentation with energy transfer usingγ-glutamylmethylamide synthetase of Methylovorus mays No.9[J].Bioscience Biotechnology&Biochemistry,2008,72(5):1206-1211.
[25]PHILIP H,HOOPER S D,KYRPIDES N C.Focus:synergistetes[J].Environmental Microbiology,2009,11(6):1327-1329.
[26]WORTHEN P L,GODE C J,GRAF J.Culture-independent characterization of the digestive-tract microbiota of the medicinal leech reveals a tripartite symbiosis[J].Applied&Environmental Microbiology,2006,72(7):4775-81.
[27]杨预展.长江中下流域草食性雁食性及肠道微生物研究[D].合肥:中国科学技术大学生命科学学院,2016.
[28]WAITE D W,TAYLOR M W.Characterizing the avian gut microbiota:membership,driving influences,and potential function[J].Frontiers in Microbiology,2014,5(4):1-12.
[29]ZHANG B,LIN Q,LIN J,et al.Effects of broodstock density and diet on reproduction and juvenile culture of the Leech,Hirudinaria manillensis,Lesson,1842[J].Aquaculture,2008,276(1/4):198-204.