【目的】研究长期不同施肥制度下黄土旱塬黑垆土氨氧化古菌群落多样性和丰度的变化,为提高黄土高原地区氮素利用效率、检测土壤质量变化提供重要依据。【方法】利用PCR-RFLP技术和Real-time PCR技术分析无肥(CK)、有机肥(M)、氮肥+有机肥(NM)、磷肥+有机肥(PM)、氮磷肥+有机肥(NPM)等5种长期施肥处理对土壤氨氧化古菌群落结构和丰度的影响。【结果】从5种施肥处理的氨氧化古菌氨单加氧酶A(amoA)的克隆文库中分别得到25、18、29、20、30种RFLP酶切类型。α多样性指数表明各施肥处理土样之间氨氧化古菌多样性差异显著,NPM处理产生的氨氧化古菌多样性最多,而M处理多样性最少。欧氏距离聚类结果表明各施肥处理与对照(CK)处理土壤中氨氧化古菌群落类型趋同性系数最大,说明长期施肥处理使氨氧化古菌群落类型产生了较大变化。各施肥处理土壤中氨氧化古菌amoA基因拷贝数不同,NPM处理的氨氧化古菌丰度最高,与其它施肥处理呈显著性差异。基于amoA氨基酸序列构建的系统发育树将5种施肥处理的主要的优势氨氧化古菌与来自土壤/淡水沉积物的未培养氨氧化古菌聚为一类。【结论】长期不同施肥改变了土壤中氨氧化古菌群落多样性和丰度。
【Objective】In order to improve the efficiency of nitrogen utilization and explicate the function of ammonia-oxidizing archaea(AOA) under the changes of soil quality in the Loess Plateau,the community structure diversity and abundance of AOA were studied.【Method】 The influence of long-term fertilization treatments including CK,M,NM,PM and NPM on soil AOA community structure diversity and amoA gene copy numbers were analyzed by restriction fragment length polymorphism(PCR-RFLP) and real-time PCR.【Result】From the clone libraries of the different fertilization treatments,there were 25,18,29,20 and 30 restriction endonuclease types,respectively.The α diversity indices indicated that there was a pronounced difference among five fertilizer treatments.The OTUs was the highest in NPM treatment and the lowest diversity in M treatment.The rescaled distance matrix tree indicated that the different fertilization had the largest convergence coefficient of AOA community types with the CK treatment soil,so the different fertilization led to significant changes of AOA communities.The amoA gene copy numbers of AOA changes were different among the treatments,whereas the highest copy numbers were detected in the NPM treatment,and had a pronounced difference with other fertilizer treatments.All preponderant sequences of AOA fell within soils/fresh water sediments based on phylogenetic tree of amoA gene amino acid sequences analysis.【Conclusion】Long-term fertilization resulted in changes of AOA community diversity and abundance.