洞庭湖岸边带沉积物氨氧化古菌的丰度、多样性及对氨氧化的贡献  

Abundance,biodiversity and contribution to ammonia oxidization of ammonia-oxidizing archaea in littoral sediments of Dongting Lake

作　　者：周磊榴[1,2] 祝贵兵[1] 王衫允[1,3] 姜博[1] 

机构地区：[1]中国科学院生态环境研究中心环境水质学国家重点实验室,北京100085 [2]中国科学院大学,北京100039 [3]哈尔滨工业大学城市水资源与水环境国家重点实验室,哈尔滨150090

出　　处：《环境科学学报》2013年第6期1741-1747,共7页Acta Scientiae Circumstantiae

基　　金：国家自然科学基金(No.21077119);国家重点基础研究发展(973)计划项目(No.2009CB421103);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.12L03ESPC);北京市科技新星计划项目(No.2011095);香港王宽诚教育基金~~

摘　　要：采用分子生物学的方法(定性/定量PCR、克隆文库)对洞庭湖岸边带沉积物中氨氧化古菌(Ammonia-Oxidizing Archaea,AOA)进行分子水平的多样性和丰度分析,并结合硝化速率潜势(Potential Nitrification Rate,PNR)剖析AOA在洞庭湖岸边带沉积物中氨氧化的作用.采用实时荧光定量PCR技术对氨氧化古菌AOA和氨氧化细菌(Ammonia-Oxidizing Bacteria,AOB)进行靶向amoA基因(氨氧化关键功能基因)的定量分析,发现古菌amoA基因的丰度为1.49×107～1.97×108copies·g-1(以干土计),高于细菌amoA基因丰度(1.52×104～2.45×106copies·g-1,以干土计)1～4个数量级.洞庭湖岸边带沉积物硝化速率潜势为1.11～6.47nmol·g-·1h-1(以N计),比细胞硝化速率为0.55～1.68fmol·cell-·1d-1(以NH3计).硝化速率潜势与古菌amoA基因丰度之间呈现正向变化趋势,而与细菌amoA基因丰度呈负向变化趋势,指示了低氨氮环境下AOA在氨氧化过程中的主导作用.生物多样性分析表明,经酶切分型后得到的22个古菌amoA基因序列以97%的相似度划分为8个独立操作单元(Operational Taxonomic Unit,OTU).系统发育分析显示,OTU1～6(14个序列)属于第一分支,OTU7～8(8个序列)属于第二分支,且均属于一个新命名的古菌类群--奇古菌门(Thaumarchaeota).With the method of molecular biology（qualitative/quantitative PCR and clone libraries）,the biodiversity and abundance of ammonia-oxidizing archaea（AOA） from the littoral sediments of Dongting Lake were analyzed at the molecular level.At the same time,the contribution of AOA to ammonia oxidization was also analyzed in combination with the potential nitrification rate（PNR）.The number of archaeal amoA gene（the key functional gene of AOA） was measured at 1.49×107-1.97×108 copies g^-1 dry soil,1-4 orders of magnitude larger than that of bacterial amoA gene（1.52×104-2.45×106 copies g^-1 dry soil）.The PNR of littoral sediments of Dongting Lake was between 1.11 and 6.47 nmol g^-1 h-1,and cell-specific nitrification rates for AOA and AOB（combined） ranged from 0.55 to 1.68 fmol cell-1 d-1.PNR correlated positively with the number of archaeal amoA gene and negatively with bacterial amoA gene,which indicated the dominant role of AOA in ammonia oxidization process in environments with low ammonium concentration.In the analysis of biodiversity,sequences sharing 97% similarity were grouped into the same operational taxonomic unit（OTU）,and 8 OTUs were obtained from 22 archaeal amoA gene sequences selected after distinguished by the restriction endonuclease.Phylogenetic analysis indicated that OTU 1-6（14 sequences） belonged to cluster 1,while OTU 7-8（8 sequences） belonged to cluster 2,and AOA in littoral sediments of Dongting Lake were all affiliated to a newly named archaea cluster ＂Thaumarchaeota＂.

关 键 词：氨氧化古菌(AOA) 多样性 丰度 岸边带沉积物 

分 类 号：X171[环境科学与工程—环境科学]