高通量测序技术研究典型湖泊岸边陆向深层土壤中厌氧氨氧化细菌的群落结构  

Community structure of anaerobic ammonia oxidation bacteria in subsurface soil from riparian zone to land in Baiyangdian Lake using high-throughput sequencing technology

作　　者：庄林杰 夏超[2,3] 田晴[1] 朱艳彬[1] 祝贵兵[2] 

机构地区：[1]东华大学环境科学与工程学院,上海201620 [2]中国科学院生态环境研究中心饮用水科学与技术重点实验室,北京100085 [3]安徽省水利水电勘测设计院勘测分院,合肥230088

出　　处：《环境科学学报》2017年第1期261-271,共11页Acta Scientiae Circumstantiae

基　　金：国家自然科学基金(No.41322012;41671471);中国科学院战略性先导科技专项(No.XDB15020303);环境模拟与污染控制国家重点联合实验室专项经费项目(No.14L01ESPC);中国科学院饮用水科学与技术重点实验室2015年度自由研究课题(No.15Z07KLDWST);中国科学院青年创新促进会资助项目~~

摘　　要：为研究具有富营养化特征的白洋淀湖泊岸边陆向深层土壤中厌氧氨氧化细菌的群落结构组成,针对厌氧氨氧化的功能基因hzs B(联氨合成酶关键基因)进行高通量测序.结果显示,白洋淀湖泊岸边陆向深层土壤共8个样品处理后共得到77082条序列,在相似度90%下聚类得到40个OTUs.多样性分析(OTUs水平)表明,在白洋淀湖泊岸边近湖岸交界处,上层土壤中厌氧氨氧化细菌的群落多样性要高于下层,而在远湖岸交界处则相反,下层土壤中厌氧氨氧化细菌的群落多样性要高于上层土壤.PCo A分析(OTUs水平)表明,近湖岸交界处采样点各不同深度土壤层之间的厌氧氨氧化细菌的群落结构更相似,而远湖岸界面处采样点各不同深度之间的厌氧氨氧化细菌的群落结构差异较大.群落结构分析表明,在白洋淀湖泊岸边陆向深层土壤中,序列隶属于Candidatus‘Brocadia anammoxidans’(34.07%)、Candidatus‘Brocadia fulgida’(24.66%)、Candidatus‘Jettenia sp.’(3.00%)、Candidatus‘Kuenenia sp.’(0.22%)、Candidatus‘Scalindua sp.’(0.08%)及与Genbank中已探明的厌氧氨氧化细菌亲缘关系较远的Anammox-like cluster(37.98%).相关性分析(p<0.10)表明,在白洋淀湖泊岸边陆向深层土壤中,厌氧氨氧化细菌α-多样性指数中丰富度Chao指数与含水率(MC)呈负相关,多样性Shannon指数与氨氮(NH+4)呈负相关,优势度Simpson指数与NH+4呈正相关;但厌氧氨氧化细菌群落组成中Candidatus‘Brocadia anammoxidans’的富集与总碳(TC)及总硫(TS)含量呈正相关.综上,本文针对厌氧氨氧化功能基因hzs B扩增进行高通量测序,一定程度上揭示了具有富营养化特征的白洋淀湖泊岸边陆向深层土壤中厌氧氨氧化细菌的多样性、群落组成及其与土壤环境因子的关系.To investigate the community structure of anaerobic ammonia oxidation（ anammox） bacteria in subsurface soil from riparian zone to land in Baiyangdian Lake with eutrophication characteristics,the high-throughput sequencing aiming at the hydrazine synthase β-subunit（ hzs B） of anammox bacteria was performed. The results showed that a total of 77082 preprocessed reads were clustered into 40 OTUs（ operational taxonomical units） at 90%identity. Diversity analysis（ based on OTUs level） shows that anammox bacteria at upper soil near shore junction in Baiyangdian Lake have higher diversity than the lower. In the far shore junction,anammox bacteria at lower soil have higher diversity than the upper soil. PCo A analysis（ based on OTUs level） shows that the community structures of anammox bacteria from different depths of soil in same site are more similar in subsurface soil near shore junction. However,there are quite large differerces of anammox bacteria community structures betweem soils from different depths in same sites far away from shore junction. Community analysis shows that the sequences in Baiyangdian riparian zone affiliated to Candidatus ‘Brocadia anammoxidans ’account for 34.07%,Candidatus‘Brocadia fulgida’account for 24.66%,Candidatus‘Jettenia sp.’account for 3.00%,Candidatus‘Scalindua sp.’and Candidatus‘Kuenenia sp.’account for 0. 08% and 0. 22% respectively; Anammox-like cluster,which have a distant relationship to explored anammox bacteria from Genbank account for 37.98%. Correlation analysis（ p〈0.10） showed Chao1 index was positively correlated with moisture content,Shannon index was negatively correlated with ammonia,Simpson index positively correlated with ammonia. However,high value of TC and TS were in favour of the enrichment of Candidatus ‘Brocadia anammoxidans’. In conclusion,this study conducted high-throughput sequencing for functional gene（ hzs B） of anaerobic ammonia oxidizing,revealed diversity,community structure of anammox bacteria and

关 键 词：厌氧氨氧化细菌 高通量测序 功能基因hzs B 深层土壤 群落结构 

分 类 号：X53[环境科学与工程—环境工程]