The two-step nitrification process is an integral part of the global nitrogen cycle, and it is accomplished by distinctly different nitrifiers. By combining DNA-based stable isotope probing (SIP) and high-throughput pyrosequencing, we present the molecular evidence for autotrophic growth of ammonia-oxidizing bacteria (AOB), ammonia-oxidizing archaea (AOA) and nitrite-oxidizing bacteria (NOB) in agricultural soil upon ammonium fertilization. Time-course incubation of SIP microcosms indicated that the amoA genes of AOB was increasingly labeled by 13 CO 2 after incubation for 3, 7 and 28 days during active nitrification, whereas labeling of the AOA amoA gene was detected to a much lesser extent only after a 28-day incubation. Phylogenetic analysis of the 13 C-labeled amoA and 16S rRNA genes revealed that the Nitrosospira cluster 3-like sequences dominate the active AOB community and that active AOA is affiliated with the moderately thermophilic Nitrososphaera gargensis from a hot spring. The higher relative frequency of Nitrospira-like NOB in the 13 C-labeled DNA suggests that it may be more actively involved in nitrite oxidation than Nitrobacter-like NOB. Furthermore, the acetylene inhibition technique showed that 13 CO 2 assimilation by AOB, AOA and NOB occurs only when ammonia oxidation is not blocked, which provides strong hints for the chemolithoautotrophy of nitrifying community in complex soil environments. These results show that the microbial community of AOB and NOB dominates the nitrification process in the agricultural soil tested. The ISME Journal (2011) 5, 1226-1236; doi:10.1038/ismej.2011.5; published online 17 February 2011Subject Category: microbial ecology and functional diversity of natural habitats
A balanced fertilization can increase crop yields partly due to stimulated microbial activities and growths. In this study, we investigated arbuscular mycorrhizal fungi (AMF) in arable soils to determine the optimal practices for an effective fertilization. We used pyrosequencing-based approach to study AMF diversity, as well as their responses to different long-term (>20 years) fertilizations, including OM (organic manure) and mix chemical fertilizers of NP (nitrogen-phosphorus), NK (nitrogen-potassium), and NPK (nitrogen-phosphorus-potassium). Results revealed that 124,998 of 18S rRNA gene fragments were dominated by Glomeromycota with 59,611 sequences, generating 70 operational taxonomic units (OTUs), of which the three largest families were Glomeraceae, Gigasporaceae and Acaulosporaceae. In Control and NK plots, AMF diversity and richness significantly decreased under long-term P fertilizations, such as NP, NPK, and OM. Concomitantly, the AMF community structure shifted. Supported by canonical correspondence analysis, we hereby propose that long-term balanced fertilization, especially P fertilizer with additional N fertilizer, helps the build-up of soil nutrients. Consequently, some AMF community constituents are sacrificed, propelled by the self-regulation of plant-AMF-microbes system, resulting in an agro-ecosystem with a better sustainability. This knowledge would be valuable toward better understandings of AMF community in agro-ecosystem, and long-term ecosystem benefits of the balanced fertilization.
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