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Precipitation changes altered soil heterotrophic respiration, but the underlying microbial mechanisms remain rarely studied. This study conducted three-year switchgrass (Panicum virgatum L.) mesocosm experiment to investigate soil heterotrophic respiratory responses to altered precipitation. Five treatments were considered, including ambient precipitation (P0), two wet treatments (P+33 and P+50: 33% and 50% enhancement relative to P0), and two drought treatments (P-33 and P-50: 33% and 50% reduction relative to P0). The plant’s aboveground biomass (AGB), soil organic carbon (SOC), total nitrogen (TN), microbial biomass carbon (MBC), heterotrophic respiration (Rs), biomass-specific respiration (Rss: respiration per unit of microbial biomass as a reciprocal index of microbial growth efficiency), and extracellular enzymes activities (EEAs) were quantified in soil samples (0–15 cm). Despite significantly different soil moisture contents among treatments, results showed no impact of precipitation treatments on SOC and TN. Increasing precipitation had no effect, but decreasing precipitation significantly reduced plant AGB. Relative to P0, P+33 significantly increased Rs by more than 3-fold and caused no changes in MBC, leading to significantly higher Rss (P < 0.05). P+33 also significantly increased hydrolytic enzyme activities associated with labile carbon acquisition (Cacq) by 115%. The only significant effect of drought treatments was the decreased β-D-cellobiosidase (CBH) and peroxidase (PEO) under P-33. Nonparametric analyses corroborated the strong influences of moisture and CBH on the enhanced precipitation, which stimulated soil respiratory carbon loss, likely driven by both elevated hydrolase activities and reduced microbial growth efficiency. However, the less sensitive drought effects suggested potential microbial tolerance to water deficiency despite depressed plant growth. This study informs the likely decoupled impacts of microbes and plants on soil heterotrophic respiration under changing precipitation in the switchgrass mesocosm experiment. 

Abstract Nitrogen (N) fertilization significantly affects soil extracellular oxidases, agents responsible for decomposition of slow turnover and recalcitrant soil organic carbon (SOC; e.g., lignin), and consequently influences soil carbon sequestration capacity. However, it remains unclear how soil oxidases mediate SOC sequestration under N fertilization, and whether these effects co‐vary with plant type (e.g., bioenergy crop species). Using a spatially explicit design and intensive soil sampling strategy under three fertilization treatments in switchgrass (SG:Panicum virgatumL.) and gamagrass (GG:Tripsacum dactyloidesL.) croplands, we quantified the activities of polyphenolic oxidase (PHO), peroxidase (PER), and their sum associated with recalcitrant C acquisition (OX). The fertilization treatments included no N fertilizer input (NN), low N input (LN: 84 kg N ha⁻¹ year⁻¹in urea), and high N input (HN: 168 kg N ha⁻¹ year⁻¹in urea). Besides correlations between soil oxidases and SOC (formerly published), both descriptive and geostatistical approaches were applied to evaluate the effects of N fertilization and crop type on soil oxidases activities and their spatial distributions. Results showed significantly negative correlations between soil oxidase activities and SOC across all treatments. The negative relationship of soil oxidases and SOC was also evident under N fertilization. First, LN significantly depressed oxidases in both mean activities and spatial heterogeneity, which corresponded to increased SOC in SG (though by 5.4%). LN slightly influenced oxidases activities and their spatial heterogeneity, consistent with insignificant changes of SOC in GG. Second, HN showed trends of decrease in soil oxidase activities, which aligned with the significantly enhanced SOC in both croplands. Overall, this study demonstrated that soil oxidase activities acted as sensitive and negative mediators of SOC sequestration in bioenergy croplands and optimizing fertilizer use particularly in switchgrass cropland can improve for both carbon sequestration and environmental benefit.