高通量测序分析新疆沼液中发酵微生物的多样性（英文）
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摘要
利用高通量测序技术研究了5种新疆不同地区的沼气池(A08,A09,A010,A011和A012)的微生物群落结构和多样性。获得5个沼气池细菌和真菌的物种注释的运算分类单位(OTU)数目,其中细菌OTU数为2105,其相同的OTU数为225;真菌OTU的总数为2224,其相同的OTU数为178。以牛粪为底物的沼液样品A08,A09,A010细菌门相对丰度由高到低分别是拟杆菌门、厚壁菌门和变形菌门。以鸡粪为底物的沼液A011的细菌门主要是拟杆菌门和厚壁菌门,其相对丰度分别为58.5%和23.8%。以猪粪为底物的样品A012细菌门相对丰度由高到低分别是厚壁菌门、拟杆菌门和变形菌门。5种沼液样品中的主要真菌门是子囊菌门和担子菌门,其中子囊菌门占多数,相对丰度为72.7%～83.3%,担子菌门的相对丰度为4.3%～7.3%。新疆不同地区和不同底物的沼气微生物种类有很大不同,研究其不同微生物结构为制备高效沼气发酵菌剂提供了参考。
The microbial varieties of 5 biogas digesters(A08, A09, A010, A011, and A012) were investigated using high-throughput sequencing in different regions in Xinjiang. The operational taxonomic units(OTU) number of bacterial in the OTU cluster analysis of 5 biogas digesters was 2 105, and their identical OTU number was 225. The total OTU number of fungal was 2 224, and their identical OTU number was 178.The main types of biogas slurry samples, A08, A09, and A010, with cow manure as substrate were Bacteroidetes, Firmicutes, and Proteobacteria. The sample A011 with chicken manure as the substrate was mainly Bacteroides and Firmicutes, and the relative abundance was 58.5% and 23.8%, respectively. The sample A012 with pig manure as substrate was similar to the sample A09 with cow manure as substrate. The first dominant group was Firmicutes, the second dominant group was Bacteroides, and the third one was Proteobacteria. The main fungi in the 5 biogas slurry samples were Ascomycota and Basidiomycota at the phylum level, in which the relative abundance of Ascomycota accounted for the majority, reached 72.7%–83.3%, with a relatively abundant level of Basidiomycetes at 4.3%–7.3%. The Xinjiang biogas digesters contained an extremely abundant number of microbial strains, and the microbial distributions of the different samples were different, which provided theory evidence for the preparation of high efficiency biogas fermenting agents.
The microbial varieties of 5 biogas digesters(A08, A09, A010, A011, and A012) were investigated using high-throughput sequencing in different regions in Xinjiang. The operational taxonomic units(OTU) number of bacterial in the OTU cluster analysis of 5 biogas digesters was 2 105, and their identical OTU number was 225. The total OTU number of fungal was 2 224, and their identical OTU number was 178.The main types of biogas slurry samples, A08, A09, and A010, with cow manure as substrate were Bacteroidetes, Firmicutes, and Proteobacteria. The sample A011 with chicken manure as the substrate was mainly Bacteroides and Firmicutes, and the relative abundance was 58.5% and 23.8%, respectively. The sample A012 with pig manure as substrate was similar to the sample A09 with cow manure as substrate. The first dominant group was Firmicutes, the second dominant group was Bacteroides, and the third one was Proteobacteria. The main fungi in the 5 biogas slurry samples were Ascomycota and Basidiomycota at the phylum level, in which the relative abundance of Ascomycota accounted for the majority, reached 72.7%–83.3%, with a relatively abundant level of Basidiomycetes at 4.3%–7.3%. The Xinjiang biogas digesters contained an extremely abundant number of microbial strains, and the microbial distributions of the different samples were different, which provided theory evidence for the preparation of high efficiency biogas fermenting agents.
引文
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[2]Ruane J,Sonnino A,Agostini A.Bioenergy and the potential contribution of agricultural biotechnologies in developing countries[J].Biomass and Bioenergy,2010,34(10):1427-1439.
[3]Bouallagui H,Lahdheb H,Romdan E B,et al.Improvement of fruit and vegetable waste anaerobic digestion performance and stability with co-substrates addition[J].Journal of Environmental Management,2009,90(5):1844-1849.
[4]LiaoCuiping,Yan Yongjie,Wu Chuangzhi,et al.Study on the distribution and quantity of biomass residues resource in china[J].Biomass and Bioenergy,2004,27(2):111-117.
[5]Weiland P.Biogas production:Current state and perspectives[J].Applied Microbiology and Biotechnology,2010,85(4):849-860.
[6]Dinuccio E,Balsari P,Gioelli F,et al.Evaluation of the biogas productivity potential of some italianagro-industrial biomasses[J].Bioresource Technology,2010,101(10):3780-3783.
[7]Sun Shirong,Guo Zi,Yue Jinquan.The status and evaluation of rice straw resource utilization in China[J].Agriculture and technology,2015,35(17):20-23.
[8]Liu Hui,Jiang Gaoming,Zhuang Huiyong,et al.Distribution,utilization structure and potential of biomass resources in rural China:With special references of crop residues[J].Renewable and Sustainable Energy Reviews,2008,12(5):1402-1418.
[9]Grover P,Mishra S.Biomass briquetting:Technology and practices[M].Bangkok:The FAO Regional Wood Energy Development Programme in Asia,1996.
[10]Xiong Chengyong,Li Jian,Huang Lihong.Simple analyse on straw biogas utilization for household digester[J].Renewable Energy,2003(3):44-45.
[11]Werther J,Saenger M,Hartge E,et al.Combustion of agricultural residues[J].Progress in Energy and Combustion Science,2000,26(1):1-27.
[12]Lin Bin.The design of swine manure treatment techniques on intensive feeding swine farm[J].Fujian Journal of Agricultural Sciences,2006,21(4):420-424.
[13]Zhang Cuili,Li Yibing,Pu Dongsheng.Biogas yield and its relations with the duration and temperature of mixed anaerobic fermentation of livestock dungs and wheat straw[J].Chinese Journal of Applied Ecology,2008,19(8):1818-1822.
[14]Lin Daiyan,Ye Meifeng,Wu Feilong,et al.Research on recycling model construction and technology integration of feces from large-scale pig farm[J].Journal of AgroEnvironment Science,2010,29(2):386-391.
[15]Chen Xiaohua,Zhu Hongguang.Research progress and prospect on producing biogas from crop straws[J].Transactions of the Chinese Society of Agricultural Engineering(Transactions of the CSAE),2007,23(3):279-283.
[16]Michal Gaworski,S?awomir Jablonski,Izabela PawlaczykGraja,et al.Enhancing biogas plant production using pig manure and corn silage by adding wheat straw processed with liquid hot water and steam explosion[J].Biotechnology for Biofuels,2017,10(1):259-271.
[17]Marcin Zieliński,Marcin D?bowski,Marta Kisielewska,et al.Comparison of ultrasonic and hydrothermal cavitation pretreatments of cattle manure mixed with straw wheat on fermentative biogas production[J].Waste Biomass Valor,2017,27(7):1-8.
[18]Luo Lina,Gong Weijia,Qin Liyuan,et al.Influence of liquid-and solid-state coupling anaerobic digestion process on methane production of cow manure and rice straw[J].Journal of Material Cycles and Waste Management,2018,20(3):1804-1812.
[19]Xi Yanhua,Chen Huicai,Zhang Liping,et al.Separation and identification of a hydrogen-producing acetogens strain ZR-1and its acetic production characters[J].Microbiology China,38(2):181-186.
[20]Li Jie,Yu Jihua,Feng Zhi,et al.Effect of different microbial inoculants on aerobic composting of cow manure[J].Journal of Arid Land Resources and Environment,2014,28(2):109-113.
[21]Shi Xiaoke,Xie Jianming,Feng Zhi,et al.Effects of three microbial agents on high temperature aerobic composting of sheep manure[J].Chinese Agricultural Science Bulletin,2015,31(2):45-48.
[22]Wang Daoze,Xie Guoxiong,Li Dan,et al.Application of microorganism agents in the composting of chicken manure[J].Acta Agriculturae Zhejiangensis,2013,25(5):1074-1078.
[23]Shi Xiaoshuang,Liu Deli,Lang Zhihong,et al.A preliminary application of PCR-DGGE in microbial community structure analysis of rural household biogas digesters[J].Journal of Shandong Normal University:Natural Science,2007,22(2):120-122.
[24]Cui Zongjun,Gong Xiaoyan,Li Guoxue.Application of denaturing gradient gel electrophoresis on sturdy of microbe in composting process[J].Microbiology China,2004,31(5):116-119.
[25]Qin Nan,Li Dongfang,Yang Ruifu.Next-generation sequencing technologies and the application in microbiology:Areview[J].Acta Microbiologica Sinica,2011,51(4):445-457.
[26]Li Qinggang,Tao Li.High-throughput sequencing technology and its application in life science[J].Animal Husbandry and Feed Science,2012,33(2):25-28.
[27]Gans J,Wolinsky M,Dunbar J.Computational improvements reveal great bacterial diversity and high metal toxicity in soil[J].Science,2005,309(5739):1387-1390.
[28]Wang Liang.Manufacture and Application of Microbial Diversity in Aerobic Composting of cattle manure[D].Beijing:Beijing Forestry University,2012.
[29]Liu Chi,Li Jiabao,Rui Junpeng,et al.The applications of the16s rRNA gene in microbial ecology:Current situation and problems[J].Acta EcologicaSinica,2015,35(9):1-25.
[30]Song Zilin,Zhang Chao.Anaerobic codigestion of pretreated wheat straw with cattle manure and analysis of the microbial community[J].Bioresource Technology,2015,186(6):128-135.
[31]Feng Jiayu,Li Yeqing,Zhang Enlan,et al.Solid-state co-digestion of NaOH-pretreated corn straw and chicken manure under mesophilic condition[J].Waste and Biomass Valorization,2018,9(6):1027-1035.
[32]Masella,A P,Bartram A K,Truszkowski J M,et al.PANDAseq:Paired-end assembler for illumina sequences[J].BMCBioinformatics,2012,13:31.
[33]Caporaso J G,Kuczynski J,Stombaugh J,et al.QIIMEallows analysis of high-throughput community sequencing data[J].Nature Methods,2010,7(5):335-336.
[34]Wang Yu,Sheng Huafang,He Yan,et al.Comparison of the levels of bacterial diversity in freshwater,intertidal wetland,and marine sediments by using millions of illumina tags[J].Applied and Environmental Microbiology,2012,78:8264-8271.
[35]Grice E A,Kong H H,Conlan S,et al.Topographical and temporal diversity of the human skin microbiome[J].Science,2009,324(5931):1190-1192.
[36]Lozupone C,Knight R.UniFrac:A new phylogenetic method for comparing microbial communities[J].Applied and Environmental Microbiology,2005,71(12):8228-8235.
[37]Jiang Xiaotao,Peng Xin,Deng Guanhua,et al.Illumina sequencing of 16s rRNA tag revealed spatial variations of bacterial communities in a mangrove wetland[J].Microbial Ecology,2013,66(1):96-104.
[38]Rundell E A,Banta L M,Ward D V,et al.16s rRNA gene survey of microbial communities in winogradsky columns[J].PLoS One,2014(9):104-134.