Broad-spectrum fungicide chlorothalonil (CTN) is successively applied into intensive agriculture soil. However, the impacts of successive CTN applications on soil nitrification and related microorganisms remain poorly understood. A microcosm study was conducted to reveal the effects of successive CTN applications on soil nitrification and functional genes involved in soil nitrogen (N) cycling. The CTN at the dosages of 5 mg kg dry soil (RD) and 25 mg kg dry soil (5RD) was successively applied into the test soil at 7-day intervals which resulted in the accumulations of CTN residues. After 28 days of incubation, CTN residues in the RD and 5RD treatments were 3.14 and 69.7 mg kg dry soil respectively. Net nitrification rates in the RD and 5RD treatments were lower than that obtained from the blank control (CK). Real-time PCR analysis revealed that AOA and AOB amoA gene abundances were significantly decreased by CTN applications. Moreover, CTN applications also discrepantly decreased the abundances of functional genes involved in soil denitrification, with the exception of nosZ gene. Principal component analysis further supported the observation that successive CTN applications could result in enhanced ecological toxicity.
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