Abstract
Nitrate leaching is severe in greenhouse agriculture where excessive nitrogen is often applied to maintain high crop productivities. Carbon amendment in the subsoil, where denitrification is limited by the availability of carbon, might mitigate nitrate leaching. In this study, we investigated the effects of carbon amendment in the subsoil on nitrate leaching and the emission of greenhouse gases (CH 4 and N 2 O) emissions using a soil column experiment. Diversity and abundance of total and nirS-, nirK- , and nosZ -type denitrifying bacteria were investigated by high throughput sequencing of PCR amplicons and quantitative real-time PCR. The amounts of nitrate leaching were >39% less in the treatments with carbon amendment than in the non-amended control without fertilization or in treatments fertilized by two doses (1600 or 3200 kg N ha - 1 ) of ammonia or nitrate. No effects of carbon amendment on the emissions of CH 4 or N 2 O were observed. The total N content in the subsoil zone with carbon amendment increased from 20.74% to 70.54%. Strikingly, the abundance of nirS , nosZ and 16S rRNA was higher in the treatment than the corresponding controls while no significant effects were detected for nirK . Carbon amendment rather than fertilization was the primary factor that influenced the community composition of the three denitrifying bacterial communities and explained 14%, 10%, and 4% of the variation in the community of nosZ, nirS , and nirK, respectively. Decreased alpha-diversity and increased variability in beta-diversity were observed for the carbon amended treatment for total and denitrifying bacteria. On average, genera such as Anaerovorax, Pseudobacteroides, Magnetospirillum, Prolixibacter, Sporobacter, Ignavibacterium, Syntrophobacter, Oxobacter, Hydrogenispora, Desulfosporomusa, Mangrovibacterium, and Sporomusa were enriched more than seven times in soil amended with carbon. In summary, carbon amendment in the subsoil mitigated nitrate leaching and increased the nitrogen pool by possible activation of denitrifying and anaerobic bacterial populations.