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The toxicity of ionic liquids (ILs) on soil organisms has aroused wide attention due to their high-solubility.The present investigation focused on the toxicity of 1-octyl-3-methylimidazolium nitrate ([C8mim]NO3) on the microbial populations (bacteria,fungi,and actinomycetes),soil enzyme (urease,dehydrogenase,acid phosphatase,and β-glucosidase)activities,microbial community diversity using terminal restriction fragment length polymorphism (T-RFLP),and abundance of the ammonia monooxygenase (amoA) genes of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) using quantitative real-time polymerase chain reaction (q-PCR) in brown soil at each trial with doses of 0,1.0,5.0,and 10.0 mg/kg on days 10,20,30,and 40.The contents of [Csmim]NO3 in soil were measured using high performance liquid chromatography with recoveries of 84.3％ to 85.2％,and changed less than 10％ during the experimental period.A significant decrease was observed from the bacteria,fungi and actinomycetes populations at 10.0 mg/kg,at which the urease activity was inhibited and the β-glucosidase activity was stimulated on days 20,30,and 40.In addition,[Csmim]NO3 inhibited the dehydrogenase activity at 10 mg/kg on days 30 and 40 and the acid phosphatase activity on day 20.The diversity of the soil microbial community and the gene abundance of AOA-and AOB-amoA were also inhibited.Furthermore,the present investigation provided more scientific information for the toxicity evaluation of ILs in soil.