Department of Crop and Soil Science, 3017 ALS, Oregon State University, Corvallis, OR 97331, USA.
Appl Environ Microbiol. 2010 Dec;76(23):7691-8. doi: 10.1128/AEM.01324-10. Epub 2010 Oct 1.
A method was developed to determine the contributions of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) to the nitrification potentials (NPs) of soils taken from forest, pasture, cropped, and fallowed (19 years) lands. Soil slurries were exposed to acetylene to irreversibly inactivate ammonia monooxygenase, and upon the removal of acetylene, the recovery of nitrification potential (RNP) was monitored in the presence and absence of bacterial or eukaryotic protein synthesis inhibitors. For unknown reasons, and despite measureable NPs, RNP did not occur consistently in forest soil samples; however, pasture, cropped, and fallowed soil RNPs commenced after lags that ranged from 12 to 30 h after acetylene removal. Cropped soil RNP was completely prevented by the bacterial protein synthesis inhibitor kanamycin (800 μg/ml), whereas a combination of kanamycin plus gentamicin (800 μg/ml each) only partially prevented the RNP (60%) of fallowed soils. Pasture soil RNP was completely insensitive to either kanamycin, gentamicin, or a combination of the two. Unlike cropped soil, pasture and fallowed soil RNPs occurred at both 30°C and 40°C and without supplemental NH(4)(+) (≤ 10 μM NH(4)(+) in solution), and pasture soil RNP demonstrated ∼ 50% insensitivity to 100 μM allyl thiourea (ATU). In addition, fallowed and pasture soil RNPs were insensitive to the fungal inhibitors nystatin and azoxystrobin. This combination of properties suggests that neither fungi nor AOB contributed to pasture soil RNP and that AOA were responsible for the RNP of the pasture soils. Both AOA and AOB may contribute to RNP in fallowed soil, while RNP in cropped soils was dominated by AOB.
开发了一种方法来确定氨氧化古菌(AOA)和氨氧化细菌(AOB)对取自森林、草地、农田和休耕(19 年)土地的土壤硝化潜力(NPs)的贡献。将土壤悬浮液暴露于乙炔中以不可逆地失活氨单加氧酶,并且在存在和不存在细菌或真核生物蛋白质合成抑制剂的情况下监测硝化潜力(RNP)的恢复。由于未知原因,尽管可测量的 NPs,但森林土壤样品中 RNP 并未一致发生;然而,在乙炔去除后 12 至 30 小时的滞后时间后,草地、农田和休耕地的土壤 RNPs 开始发生。细菌蛋白质合成抑制剂卡那霉素(800μg/ml)完全阻止了农田土壤的 RNP,而卡那霉素加庆大霉素(各 800μg/ml)的组合仅部分阻止了休耕地土壤的 RNP(60%)。草地土壤 RNP 对卡那霉素、庆大霉素或两者的组合均完全不敏感。与农田土壤不同,草地和休耕地土壤的 RNP 发生在 30°C 和 40°C 且无需补充 NH(4)(+)(溶液中≤10μM NH(4)(+)),并且草地土壤 RNP 对 100μM 烯丙基硫脲(ATU)表现出约 50%的不敏感性。此外,休耕地和草地土壤的 RNP 对真菌抑制剂制霉菌素和唑菌胺酯不敏感。这些特性的组合表明,真菌和 AOB 都没有为草地土壤的 RNP 做出贡献,而 AOA 是草地土壤 RNP 的原因。AOA 和 AOB 都可能为休耕地土壤的 RNP 做出贡献,而农田土壤的 RNP 则主要由 AOB 主导。