Extracellular glycosidases in soil, produced by microorganisms, act as major agents for decomposing labile soil organic carbon (e.g., cellulose). Soil extracellular glycosidases are significantly affected by nitrogen (N) fertilization but fertilization effects on spatial distributions of soil glycosidases have not been well addressed. Whether the effects of N fertilization vary with bioenergy crop species also remains unclear. Based on a 3-year fertilization experiment in Middle Tennessee, USA, a total of 288 soil samples in topsoil (0–15 cm) were collected from two 15 m2 plots under three fertilization treatments in switchgrass (SG: Panicum virgatum L.) and gamagrass (GG: Tripsacum dactyloides L.) using a spatially explicit design. Four glycosidases, α-glucosidase (AG), β-glucosidase (BG), β-xylosidase (BX), cellobiohydrolase (CBH), and their sum associated with C acquisition (Cacq) were quantified. The three fertilization treatments were no N input (NN), low N input (LN: 84 kg N ha−1 year−1 in urea) and high N input (HN: 168 kg N ha−1 year−1 in urea). The descriptive and geostatistical approaches were used to evaluate their central tendency and spatial heterogeneity. Results showed significant interactive effects of N fertilization and crop type on BX such that LN and HN significantly enhanced BX by 14% and 44% in SG, respectively. The significant effect of crop type was identified and glycosidase activities were 15–39% higher in GG than those in SG except AG. Within-plot variances of glycosidases appeared higher in SG than GG but little differed with N fertilization due to large plot-plot variation. Spatial patterns were generally more evident in LN or HN plots than NN plots for BG in SG and CBH in GG. This study suggested that N fertilization elevated central tendency and spatial heterogeneity of glycosidase activities in surficial soil horizons and these effects however varied with crop and enzyme types. Future studies need to focus on specific enzyme in certain bioenergy cropland soil when N fertilization effect is evaluated.

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 virgatum L.) and gamagrass (GG: Tripsacum dactyloides L.) 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−1 in urea), and high N input (HN: 168 kg N ha−1 year−1 in 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.

Background Soil moisture, pH, dissolved organic carbon and nitrogen (DOC, DON) are important soil biogeochemical properties in switchgrass (SG) and gamagrass (GG) croplands. Yet their spatiotemporal patterns under nitrogen (N) fertilization have not been studied. Aims The objective of this study is to investigate the main and interactive effects of N fertilization and bioenergy crop type on central tendencies and spatial heterogeneity of soil moisture, pH, DOC and DON. Methods Based on a 3‐year long fertilization experiment in Middle Tennessee, USA, 288 samples of top horizon soils (0–15 cm) under three fertilization treatments in SG and GG croplands were collected. The fertilization treatments were no N input (NN), low N input (LN: 84 kg N ha−1 in urea) and high N input (HN: 168 kg N ha−1 in urea). Soil moisture, pH, DOC and DON were quantified. And their within‐plot variations and spatial distributions were achieved via descriptive and geostatistical methods. Results Relative to NN, LN significantly increased DOC content in SG cropland. LN also elevated within‐plot spatial heterogeneity of soil moisture, pH, DOC and DON in both croplands though GG showed more evident spatial heterogeneity than SG. Despite the pronounced patterns described above, great plot to plot variations were also revealed in each treatment. Conclusion This study informs the generally low sensitivity of spatiotemporal responses in soil biogeochemical features to fertilizer amendments in bioenergy croplands. However, the significantly positive responses of DOC under low fertilizer input informed the best practice of optimizing agricultural nutrient amendment.

The study of warming impact on soils requires a realistic and accurate representation of temperature. In laboratory incubation studies, a widely adopted method has been to render constant temperatures in multiple chambers, and via comparisons of soil responses between low-and high-temperature chambers, to derive the warming impact on soil changes. However, this commonly used method failed to imitate both the magnitude and amplitude of actual temperatures as observed in field conditions, thus potentially undermining the validity of such studies. With sophisticated environmental chambers becoming increasingly available, it is imperative to examine alternative methods of temperature control for soil incubation research. This protocol will introduce a state-of-the-art environmental chamber and demonstrate both conventional and new methods of temperature control to improve the experimental design of soil incubation. The protocol mainly comprises four steps: temperature monitoring and programming, soil collection, laboratory incubation, and warming effect comparison. One example will be presented to demonstrate different methods of temperature control and the resultant contrasting warming scenarios; that is, a constant temperature design referred to as stepwise warming (SW) and simulated in situ temperature design as gradual warming (GW), as well as their effects on soil respiration, microbial biomass, and extracellular enzyme activities. In addition, we present a strategy to diversify temperature change scenarios to meet specific climate change research needs (e.g., extreme heat). The temperature control protocol and the recommended well-tailored and diversified temperature change scenarios will assist researchers in establishing reliable and realistic soil incubation experiments in the laboratory.

The study of warming impact on soils requires a realistic and accurate representation of temperature. In laboratory incubation studies, a widely adopted method has been to render constant temperatures in multiple chambers, and via comparisons of soil responses between low-and high-temperature chambers, to derive the warming impact on soil changes. However, this commonly used method failed to imitate both the magnitude and amplitude of actual temperatures as observed in field conditions, thus potentially undermining the validity of such studies. With sophisticated environmental chambers becoming increasingly available, it is imperative to examine alternative methods of temperature control for soil incubation research. This protocol will introduce a state-of-the-art environmental chamber and demonstrate both conventional and new methods of temperature control to improve the experimental design of soil incubation. The protocol mainly comprises four steps: temperature monitoring and programming, soil collection, laboratory incubation, and warming effect comparison. One example will be presented to demonstrate different methods of temperature control and the resultant contrasting warming scenarios; that is, a constant temperature design referred to as stepwise warming (SW) and simulated in situ temperature design as gradual warming (GW), as well as their effects on soil respiration, microbial biomass, and extracellular enzyme activities. In addition, we present a strategy to diversify temperature change scenarios to meet specific climate change research needs (e.g., extreme heat). The temperature control protocol and the recommended well-tailored and diversified temperature change scenarios will assist researchers in establishing reliable and realistic soil incubation experiments in the laboratory.