This study aims to investigate the effect of sieving on ex situ soil respiration (CO 2 flux) measurements from different land use types. We collected soils (0-10 cm) from arable, grassland and woodland sites, allocated them to either sieved (4-mm mesh, freshly sieved) or intact core treatments and incubated them in gas-tight jars for 40 days at 10°C. Headspace gas was collected on days 1, 3, 17, 24, 31 and 38 and CO 2 analysed. Our results showed that sieving (4 mm) did not significantly influence soil respiration measurements, probably because micro aggregates (< 0.25 mm) remain intact after sieving. However, soils collected from grassland soil released more CO 2 compared with those collected from woodland and arable soils, irrespective of sieving treatments. The higher CO 2 from grassland soil compared with woodland and arable soils was attributed to the differences in the water holding capacity and the quantity and stoichiometry of the organic matter between the three soils. We conclude that soils sieved prior to ex situ respiration experiments provide realistic respiration measurements. This finding lends support to soil scientists planning a sampling strategy that better represents the inhomogeneity of field conditions by pooling, homogenising and sieving samples, without fear of obtaining unrepresentative CO 2 flux measurements caused by the disruption of soil architecture.

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Laboratory incubation studies evaluating the temperature sensitivity of soil respiration often use measurements of respiration taken at a constant incubation temperature from soil that has been pre-incubated at the same constant temperature. However, such constant temperature incubations do not represent the field situation where soils undergo diurnal temperature oscillations. We investigated the legacy effects of constant and diurnally oscillating temperatures on soil respiration and soil microbial community composition. A grassland soil from the United Kingdom was either incubated at a constant temperature of 5 C, 10 C, or 15 C, or diurnally oscillated between 5 C and 15 C. Soil CO 2 flux was measured by temporarily moving incubated soils from each of the above-mentioned treatments to 5 C, 10 C or 15 C, such that soils incubated under every temperature regime had CO 2 flux measured at each temperature. We hypothesised that, irrespective of measurement temperature, CO 2 emitted from the 5 C to 15 C oscillating incubation would be most like the soil incubated at 10 C. The results showed that both incubation and measurement temperatures separately influence soil respiration. Oscillations

Abstract. Predictions concerning the feedback of soil heterotrophic respiration to a warming climate often do not differentiate between the extracellular and intracellular processes involved in soil organic matter decomposition. This study examined the temperature sensitivities of intracellular and extracellular soil enzyme activities and how they are influenced by previous temperatures. We pre-incubated soils at 5 °C, 15 °C or 26 °C to acclimatise the microbial communities to different thermal regimes for 60 days before measuring potential activities of β-glucosidase and chitinase (extracellular enzymes), glucose-induced respiration (intracellular enzymes), and basal respiration at a range of assay temperatures (5 °C, 15 °C, 26 °C, 37 °C, and 45 °C). A higher pre-incubation temperature decreased soil pH and C / N ratio which exerted a strong legacy effect by decreasing β-glucosidase potential activity and respiration, but not chitinase potential activity. It is likely that this legacy effect is an indirect effect of substrate depletion rather than physiological acclimatation or genetic adaptation. There was no overall significant effect of pre-incubation temperature on temperature sensitivity of these enzymes, perhaps because of the short (60 day) duration of the pre-incubation. However, we found that the intracellular and extracellular enzyme activities differ in their temperature sensitivity and this observation differs depending on the range of temperature used for Q10 estimates of temperature sensitivity. Between 5 °C and 15 °C intracellular and extracellular enzyme activities show equal temperature sensitivity, but between 15 °C and 26 °C intracellular enzyme activity was more temperature sensitive than extracellular enzyme activity and between 26 °C and 37 °C extracellular enzyme activity was more temperature sensitive than intracellular enzyme activity. This result implies that depolymerisation of higher molecular weight carbon is more sensitive to temperature changes at higher temperatures (e.g. higher temperatures on extremely warm days) but the respiration of the generated monomers is more sensitive to temperature changes at moderate temperatures (e.g. mean daily maximum soil temperature). Therefore, since climate change predictions currently indicate that there will be a greater frequency and severity of hot summers and heatwaves, it is possible that global warming may reduce the importance of extracellular depolymerisation relative to intracellular catalytic activity as the rate limiting step of soil organic matter mineralization. We conclude that extracellular and intracellular steps are not equally sensitive to changes in soil temperature and that the previous temperature a soil is exposed to may influence the potential activity, but not temperature sensitivity, of extracellular and intracellular enzymes.