基于MiSeq测序分析酸性农作物土壤细菌群落结构与多样性  

Analysis of Bacterial Community Composition and Diversity in Acid Soil Using MiSeq Sequencing

作　　者：王怡 常彬河 刘月 梁建军[1] 薛芷筠 张泽文 何冬兰[1] WANG Yi;CHANG Binhe;LIU Yue;LIANG Jianjun;XUE Zhijun;ZHANG Zewen;HE Donglan(Hubei Engineering and Technological Research Center for Microbial Resources and Utilization,South-Central University for Nationalities,Wuhan 430074,China)

机构地区：[1]中南民族大学湖北省微生物资源与利用工程技术研究中心

出　　处：《环境科学研究》2019年第9期1575-1583,共9页Research of Environmental Sciences

基　　金：国家自然科学基金项目(No.31070087)~~

摘　　要：为分析酸性农田土壤中微生物群落结构,探寻酸化土壤的综合治理途径,以湖北恩施土家族苗族自治州利川市酸性农作物土壤为研究对象,应用Illumina MiSeq高通量测序技术,测定酸性土壤中总细菌和经驯化的可培养细菌的16S rRNA基因V3~V4变异区序列,确定不同农作物土壤中总细菌和可培养细菌的群落结构,并探讨土壤理化因子对土壤总细菌群落结构的影响.结果表明:白菜地、烟草地、山药地、草莓地、玉米地及莴苣地这6种农作物土壤中总细菌和可培养细菌在97%的相似水平下得到的OTUs(operational taxonomic units,操作分类单元)数量分别为4 008和97个.在门水平上,土壤总细菌中有12个门类的细菌相对丰度大于1%,Proteobacteria、Acidobacteria和Chloroflexi为优势菌门细菌;可培养细菌中仅有3个门类细菌相对丰度高于1%,即Firmicutes、Proteobacteria和Bacteroidetes.在OTU水平上,土壤总细菌沿pH梯度将6种土壤样品分为2类,即pH<5.5的玉米地、山药地和莴苣地和pH>5.5的烟草地、白菜地和草莓地.此外,冗余分析(RDA)结果显示,Acidobacteria的相对丰度与pH呈正相关,Firmicutes和Roteobacter的相对丰度均与w(TN)、w(TP)、w(TC)呈正相关.系统进化树分析表明,相对丰度位于前30位的细菌与亚洲地区发现细菌的16S rRNA基因序列同源性较高.研究显示,Illumina MiSeq测序能够较为全面地解析酸性土壤中总细菌和可培养细菌的多样性,驯化过程极大地减少了细菌的种类;同时,发现细菌群落结构的相似性可能因样本来源的地理距离的增加而衰减.In order to determine the microbial community composition in acidic farmland soil and explore the comprehensive treatment of acidified soil,acidic soil samples collected from Lichuan City,Enshi Tujia and Miao Autonomous Prefecture,Hubei Province,were selected as research objects,and the 16S rRNA gene V3-V4 variable region of total bacteria and culturable bacteria in each soil sample were sequenced using Illumina sequencing.Using this methodology,the effects of soil physicochemical properties on the community structure of total bacteria in soils were investigated.The results demonstrated that 4008 total bacteria and 97 OTUs culturable bacteria from Chinese cabbage,tobacco,yam,strawberry,maize and lettuce at 97%similar level were found.At phylum level,12 bacterial groups whose relative abundance were more than 1%,whilst Proteobacteria,Acidobacteria,and Chloroflexi were the dominant phyla.There were only 3 culturable phyla bacteria whose relative abundance were more than 1%,namely Firmicutes,Proteobacteria and Bacteroidetes.At OTU level,soil total bacteria could be divided into two groups across pH gradient:pH<5.5(maize,yam and lettuce fields)and pH>5.5(tobacco,cabbage and strawberry fields).RDA analysis exhibited that the relative abundance of Acidobacteria was positively related to pH,while the abundance of Firmicutes or Proteobacteria was positively correlated to w(TN),w(TP),w(TC).The phylogenetic tree showed that the top 30 OTU bacteria had higher homology with the 16S rRNA gene sequence of the bacteria found in Asia.These results showed that Illumina MiSeq sequencing can comprehensively analyze the diversity of total bacteria and culturable bacteria in acidic soils,and that the domestication process greatly reduces the species of bacteria,at the same time,the similarity of bacterial community structure may decrease with the increase of geographic distance.

关 键 词：酸性土壤 MiSeq测序 理化因子 细菌群落 可培养细菌 土壤总细菌 

分 类 号：X171[环境科学与工程—环境科学]