Department of Microbiology, Chungbuk National University, 12 Gaeshin-dong, Heungduk-gu, Cheongju 361-763, South Korea.
Appl Environ Microbiol. 2011 Dec;77(24):8635-47. doi: 10.1128/AEM.05787-11. Epub 2011 Oct 14.
Soil nitrification is an important process for agricultural productivity and environmental pollution. Though one cultivated representative of ammonia-oxidizing Archaea from soil has been described, additional representatives warrant characterization. We describe an ammonia-oxidizing archaeon (strain MY1) in a highly enriched culture derived from agricultural soil. Fluorescence in situ hybridization microscopy showed that, after 2 years of enrichment, the culture was composed of >90% archaeal cells. Clone libraries of both 16S rRNA and archaeal amoA genes featured a single sequence each. No bacterial amoA genes could be detected by PCR. A [¹³C]bicarbonate assimilation assay showed stoichiometric incorporation of ¹³C into Archaea-specific glycerol dialkyl glycerol tetraethers. Strain MY1 falls phylogenetically within crenarchaeal group I.1a; sequence comparisons to "Candidatus Nitrosopumilus maritimus" revealed 96.9% 16S rRNA and 89.2% amoA gene similarities. Completed growth assays showed strain MY1 to be chemoautotrophic, mesophilic (optimum at 25°C), neutrophilic (optimum at pH 6.5 to 7.0), and nonhalophilic (optimum at 0.2 to 0.4% salinity). Kinetic respirometry assays showed that strain MY1's affinities for ammonia and oxygen were much higher than those of ammonia-oxidizing bacteria (AOB). The yield of the greenhouse gas N₂O in the strain MY1 culture was lower but comparable to that of soil AOB. We propose that this new soil ammonia-oxidizing archaeon be designated "Candidatus Nitrosoarchaeum koreensis."
土壤硝化作用是农业生产力和环境污染的重要过程。虽然已经描述了一种来自土壤的氨氧化古菌的培养代表,但还需要对其他代表进行特征描述。我们描述了一种从农业土壤中高度富集培养的氨氧化古菌(菌株 MY1)。荧光原位杂交显微镜显示,经过 2 年的富集,培养物由>90%的古菌细胞组成。16S rRNA 和古菌 amoA 基因的克隆文库各自具有一个单一的序列。通过 PCR 无法检测到细菌 amoA 基因。[¹³C]碳酸氢盐同化测定显示,¹³C 被等量地掺入到古菌特异性甘油二烷基甘油四醚中。菌株 MY1 在泉古菌组 I.1a 中系统发育上属于内古菌;与“Candidatus Nitrosopumilus maritimus”的 16S rRNA 和 amoA 基因序列比较显示,16S rRNA 相似性为 96.9%,amoA 基因相似性为 89.2%。完成的生长测定表明,菌株 MY1是化能自养型、嗜中温型(最佳生长温度为 25°C)、嗜中性(最佳 pH 值为 6.5 至 7.0)和非嗜盐型(最佳盐度为 0.2 至 0.4%)。动力学呼吸测定表明,菌株 MY1对氨和氧的亲和力远高于氨氧化细菌(AOB)。在菌株 MY1 培养物中温室气体 N₂O 的生成量较低,但与土壤 AOB 相当。我们建议将这种新的土壤氨氧化古菌命名为“Candidatus Nitrosoarchaeum koreensis”。