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Nitrogen (N) deposition can influence carbon cycling of terrestrial ecosystems. However, a general recognition of how soil microorganisms respond to increasing N deposition is not yet reached. We explored soil microbial responses to two levels of N addition (2.5 and 5 gN m(-2) yr(-1)) in interplant soil and beneath shrubs of Haloxylon ammodendron and their consequences to soil respiration in the Gurbantunggut Desert, northwestern China from 2011 to 2013. Microbial biomass and respiration were significantly higher beneath H. ammodendron than in interplant soil. The responses of microbial biomass carbon (MBC) and microbial respiration (MR) showed opposite responses to N addition in interplant and beneath H. ammodendron. N addition slightly increased MBC and MR in interplant soil and decreased them beneath H. ammodendron, with a significant inhibition only in 2012. N addition had no impacts on the total microbial physiological activity, but N addition decreased the labile carbon substrate utilization beneath H. ammodendron when N addition level was high. Phospholipid fatty acid (PLFA) analysis showed that N addition did not alter the soil microbial community structure as evidenced by the similar ratios of fungal to bacterial PLFAs and gram-negative to gram-positive bacterial PLFAs. Microbial biomass and respiration showed close correlations with soil water content and dissolved carbon, and they were independent of soil inorganic nitrogen across three years. Our study suggests that N addition effects on soil microorganisms and carbon emission are dependent on the respiratory substrates and water availability in the desert ecosystem. (C) 2015 Elsevier B.V. All rights reserved.