由氨氧化微生物驱动的氨氧化过程是硝化作用的限速步骤,在土壤氮素循环过程中扮演着重要角色.以湖南省宁乡县长达30 a定位试验水稻土壤为研究对象,采用荧光定量PCR和Illumina Mi Seq高通量测序分析方法,以amoA基因为靶标,研究了4种施肥...由氨氧化微生物驱动的氨氧化过程是硝化作用的限速步骤,在土壤氮素循环过程中扮演着重要角色.以湖南省宁乡县长达30 a定位试验水稻土壤为研究对象,采用荧光定量PCR和Illumina Mi Seq高通量测序分析方法,以amoA基因为靶标,研究了4种施肥制度[不施肥(CK)、化肥(CF)、70%化肥+30%有机肥(CFM_1)和40%化肥+60%有机肥(CFM_2)]水稻土壤氨氧化微生物的数量和群落结构变化.结果表明:不同施肥处理氨氧化古菌(AOA)和氨氧化细菌(AOB)amoA基因拷贝数分别为3.09×10~7～8.37×10~7和1.04×10~7～7.03×10~7copies·g^(-1)干土.施肥显著提高了AOA和AOB数量,但处理CFM_2中AOB数量与CK差异不显著.有机肥配施比例对AOB群落α多样性指数的影响强于AOA,处理CFM_1中AOA群落的多样性指数(Shannon)和AOB群落的丰富度指数(ACE和Chao1)均显著高于CK.奇古菌门和泉古菌门是AOA群落的优势门类群,占AOA amoA基因总序列的83.4%;亚硝化螺菌属、environmental_samples_norank、Bacteria_unclassified和Nitrosomonadales_unclassified是AOB群落的优势属类群,占AOB amoA基因总序列的97.8%.维恩分析结果显示,有机肥配施比例对AOB群落操作分类单元(OTU)数量的影响强于AOA,但对各处理共有AOA和AOB amoA基因序列条数的影响均较小.冗余分析结果显示,不同施肥处理AOB群落结构差异强于AOA,且所有土壤理化性质均与AOA和AOB群落结构存在显著相关关系.综上可知:有机肥配施比例显著改变了AOA和AOB数量、多样性和群落结构,配施30%有机肥时,AOA群落的Shannon指数最高,AOB群落数量、ACE和Chao1指数均最高.研究结果可为进一步探讨农业系统中氨氧化微生物对不同施肥制度的响应机制及其在氮素转化中的作用提供科学依据.展开更多
We investigated the communities of ammonia-oxidizing bacteria （AOB） in activated sludge collected from eight wastewater treatment systems using polymerase chain reaction （PCR） followed by terminal restriction frag...We investigated the communities of ammonia-oxidizing bacteria （AOB） in activated sludge collected from eight wastewater treatment systems using polymerase chain reaction （PCR） followed by terminal restriction fragment length polymorphism （T-RFLP）, cloning, and sequencing of the α-subunit of the ammonia monooxygenase gene （amoA）. The T-RFLP fingerprint analyses showed that different wastewater treatment systems harbored distinct AOB communities. However, there was no remarkable difference among the AOB T- RFLP profiles from different parts of the same system. The T-RFLP fingerprints showed that a full-scale wastewater treatment plant （WWTP） contained a larger number of dominant AOB species than a pilot-scale reactor. The source of influent affected the AOB community, and the WWTPs treating domestic wastewater contained a higher AOB diversity than those receiving mixed domestic and industrial wastewater. However, the AOB community structure was little affected by the treatment process in this study. Phylogenetic analysis of the cloned amoA genes clearly indicated that all the dominant AOB in the systems was closely related to Nitrosomonas spp. not to Nitrosospira spp. Members of the Nitrosomonas oligotropha and Nitrosomonas communis clusters were found in all samples, while members of Nitrosomonas europaea cluster occurred in some systems.展开更多
【目的】以艾比湖湿地盐节木(Halocnemum strobilaceum)、芦苇(Reed)及盐角草(Salicornia)三种植物为对象,研究其根际土壤氨氧化细菌(AOB)的群落多样性。【方法】利用PCR-RFLP的方法,构建氨氧化细菌amo A基因克隆文库,进行系统发育分析...【目的】以艾比湖湿地盐节木(Halocnemum strobilaceum)、芦苇(Reed)及盐角草(Salicornia)三种植物为对象,研究其根际土壤氨氧化细菌(AOB)的群落多样性。【方法】利用PCR-RFLP的方法,构建氨氧化细菌amo A基因克隆文库,进行系统发育分析。结合三种植物根际理化因子特点,探讨三种植物根际AOB群落结构组成的特点。【结果】通过序列多态性分析表明,三种植物根际土壤AOB amo A基因克隆文库中的所有序列均属于β亚纲(β-Proteobacteria)中的亚硝化单胞菌属(Nitrosomonas)及Nitrosomonas-like,未发现亚硝化螺菌属(Nitrosospira)。三个AOB amo A基因克隆文库分别包括9个OTUs、12个OTUs和7个OTUs,其文库覆盖度均达99%以上,代表性强。三个文库的丰富度、Chao1指数、ACE指数、Shannon指数均为R>H>S。【结论】三种植物的AOB多样性为芦苇>盐节木>盐角草,并且其最优氨氧化菌群各不相同。本研究为系统认识艾比湖湿地植物根际氨氧化细菌群落多样性和结构组成提供了基础。展开更多
基金supported by the Key Projects in National Science & Technology Pillar Program during the Eleventh Five-Year Plan Period (No.2006BAC19B01-02)the Mega-projects of Science Research for Water (No.2008ZX07313-3)the Program of Introducing Talents of Discipline to Universities
文摘We investigated the communities of ammonia-oxidizing bacteria （AOB） in activated sludge collected from eight wastewater treatment systems using polymerase chain reaction （PCR） followed by terminal restriction fragment length polymorphism （T-RFLP）, cloning, and sequencing of the α-subunit of the ammonia monooxygenase gene （amoA）. The T-RFLP fingerprint analyses showed that different wastewater treatment systems harbored distinct AOB communities. However, there was no remarkable difference among the AOB T- RFLP profiles from different parts of the same system. The T-RFLP fingerprints showed that a full-scale wastewater treatment plant （WWTP） contained a larger number of dominant AOB species than a pilot-scale reactor. The source of influent affected the AOB community, and the WWTPs treating domestic wastewater contained a higher AOB diversity than those receiving mixed domestic and industrial wastewater. However, the AOB community structure was little affected by the treatment process in this study. Phylogenetic analysis of the cloned amoA genes clearly indicated that all the dominant AOB in the systems was closely related to Nitrosomonas spp. not to Nitrosospira spp. Members of the Nitrosomonas oligotropha and Nitrosomonas communis clusters were found in all samples, while members of Nitrosomonas europaea cluster occurred in some systems.
文摘【目的】以艾比湖湿地盐节木(Halocnemum strobilaceum)、芦苇(Reed)及盐角草(Salicornia)三种植物为对象,研究其根际土壤氨氧化细菌(AOB)的群落多样性。【方法】利用PCR-RFLP的方法,构建氨氧化细菌amo A基因克隆文库,进行系统发育分析。结合三种植物根际理化因子特点,探讨三种植物根际AOB群落结构组成的特点。【结果】通过序列多态性分析表明,三种植物根际土壤AOB amo A基因克隆文库中的所有序列均属于β亚纲(β-Proteobacteria)中的亚硝化单胞菌属(Nitrosomonas)及Nitrosomonas-like,未发现亚硝化螺菌属(Nitrosospira)。三个AOB amo A基因克隆文库分别包括9个OTUs、12个OTUs和7个OTUs,其文库覆盖度均达99%以上,代表性强。三个文库的丰富度、Chao1指数、ACE指数、Shannon指数均为R>H>S。【结论】三种植物的AOB多样性为芦苇>盐节木>盐角草,并且其最优氨氧化菌群各不相同。本研究为系统认识艾比湖湿地植物根际氨氧化细菌群落多样性和结构组成提供了基础。
