Kemnitz Dana, Kolb Steffen, Conrad Ralf
Department of Biogeochemistry, Max-Planck-Institute for Terrestrial Microbiology, Marburg, Germany.
FEMS Microbiol Ecol. 2007 Jun;60(3):442-8. doi: 10.1111/j.1574-6941.2007.00310.x. Epub 2007 Mar 28.
The objective of the study was to elucidate the depth distribution and community composition of Archaea in a temperate acidic forest soil. Numbers of Archaea and Bacteria were measured in the upper 18 cm of the soil, and soil cores were sampled on two separate occasions using quantitative PCR targeting 16S rRNA genes. Maximum numbers of Archaea were 0.6-3.8 x 10(8) 16S rRNA genes per gram of dry soil. Numbers of Bacteria were generally higher, but Archaea always accounted for a high percentage of the total gene numbers (12-38%). The archaeal community structure was analysed by the construction of clone libraries and by terminal restriction length polymorphism (T-RFLP) using the same Archaea-specific primers. With the reverse primer labelled, T-RFLP analysis led to the detection of four T-RFs. Three had lengths of 83, 185 and 218 bp and corresponded to uncultured Crenarchaeota. One (447 bp) was assigned to Thermoplasmales. Labelling of the forward primer allowed further separation of the T-RF into Crenarchaeota Group I.1c and Group I.1b, and indicated that Crenarchaeota of the Group I.1c were the predominant 16S rRNA genotype (<or=85%) in the soil. The abundance of Archaea and concentration of ammonia and nitrate decreased with soil depth. Hence it is unclear if the detected Crenarchaeota Group I.1c participated in ammonia oxidation or had another phenotype.
本研究的目的是阐明温带酸性森林土壤中古菌的深度分布和群落组成。在土壤上部18厘米处测量古菌和细菌的数量,并使用靶向16S rRNA基因的定量PCR在两个不同的时间采集土芯样本。古菌的最大数量为每克干土0.6 - 3.8×10⁸个16S rRNA基因。细菌数量通常更高,但古菌在总基因数量中始终占很高的比例(12 - 38%)。使用相同的古菌特异性引物,通过构建克隆文库和末端限制性片段长度多态性(T - RFLP)分析古菌群落结构。用反向引物标记后,T - RFLP分析检测到四个末端限制性片段(T - RF)。三个片段长度分别为83、185和218 bp,对应未培养的泉古菌门。一个(447 bp)归属于嗜热栖热菌目。用正向引物标记可将T - RF进一步分离为泉古菌门I.1c组和I.1b组,并表明I.1c组泉古菌是土壤中主要的16S rRNA基因型(≤85%)。古菌的丰度以及氨和硝酸盐的浓度随土壤深度而降低。因此,尚不清楚检测到的泉古菌门I.1c组是否参与氨氧化或具有其他表型。