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Abstract Grazing intensity and global warming are expected to increase in the forthcoming decades, with uncertain consequences for their interaction on grassland ecosystems and their functions. We investigated the effects of warming, grazing and their interaction in a factorial warming (+1.2–1.7 °C) and grazing (moderate intensity with ca. 50% vegetation consumption) experiment in a Tibetan alpine meadow on microbial communities by studying functional genes involved in soil carbon and nitrogen cycles, using GeoChip technology. Our results showed that microbial functional gene structure and abundances were largely affected by the interactive effect of grazing and warming, rather than the main effect of warming or grazing. Compared to the control, grazing alone significantly increased the functional gene alpha diversity, changed the overall functional community structure, and increased the abundances of C fixation, C degradation, N mineralization and denitrification genes, likely due to the stimulating impact of urine and dung deposition. Warming alone did not change these microbial properties, possibly related to the unchanged soil nutrient status. Despite an increase in soil NO 3 − concentrations and the deposition of urine and dung, the combined treatment did not change functional gene alpha diversity, community structure, or C/N cycling gene abundances, possibly resulting from the limiting effect of water depletion in the combined treatment. Our study revealed antagonistic interactions between warming and grazing on microbial functional gene structure and abundances, which remained stable under the moderate intensity of grazing in future warming scenario in the Tibetan alpine meadow, raising potentially important implications for predicting future soil carbon and nitrogen processes in these systems. Highlights • Grazing alone significantly increased abundances of some key soil C/N cycling genes. • Warming alone had no significant effects on them. • Functional gene structure and abundances remained stable in the combined treatment. • Antagonistic interaction between warming and grazing may result from water depletion. [ABSTRACT FROM AUTHOR]

The nitrogen (N) cycle is a collection of important biogeochemical pathways mediated by microbial communities and is an important constraint in response to elevated CO 2 in many terrestrial ecosystems. Previous studies attempting to relate soil N cycling to microbial genetic data mainly focused on a few gene families by PCR, protein assays and functional gene arrays, leaving the taxonomic and functional composition of soil microorganisms involved in the whole N cycle less understood. In this study, 24 soil samples were collected from the long-term experimental site, BioCON, in 2009. A shotgun metagenome sequencing approach was employed to survey the microbial gene families involved in soil N cycle in the grassland that had been exposed to elevated CO 2 (eCO 2 ) for >12 years. In addition to evaluating the responses of major N cycling gene families to long-term eCO 2 , we also aimed to characterize the taxonomic and functional composition of these gene families involved in soil N transformations. At the taxonomic level, organic N metabolism and nitrate reduction had the most diverse microbial species involved. The distinct taxonomic composition of different N cycling processes suggested that the complex N cycle in natural ecosystems was a result of multiple processes by many different microorganisms. Belowground microbial communities that mediate N cycling responded to eCO 2 in several different ways, including through stimulated abundances of the gene families related with organic decomposition, dissimilatory nitrate reduction, and N 2 fixation, and suppressed abundances of the gene families in glutamine synthesis and anammox. This study provides a genetic basis of the microorganisms involved in key processes in the N cycle in complex ecosystems, and shows that long-term eCO 2 selectively affects N cycling pathways instead of tuning up every process. [ABSTRACT FROM AUTHOR]