The mechanisms behind the (13)C enrichment of organic matter with increasing soil depth in forests are unclear. To determine if (13)C discrimination during respiration could contribute to this pattern, we compared delta(13)C signatures of respired CO(2) from sieved mineral soil, litter layer and litterfall with measurements of delta(13)C and delta(15)N of mineral soil, litter layer, litterfall, roots and fungal mycelia sampled from a 68-year-old Norway spruce forest stand planted on previously cultivated land. Because the land was subjected to ploughing before establishment of the forest stand, shifts in delta(13)C in the top 20 cm reflect processes that have been active since the beginning of the reforestation process. As (13)C-depleted organic matter accumulated in the upper soil, a 1.0 per thousand delta(13)C gradient from -28.5 per thousand in the litter layer to -27.6 per thousand at a depth of 2-6 cm was formed. This can be explained by the 1 per thousand drop in delta(13)C of atmospheric CO(2) since the beginning of reforestation together with the mixing of new C (forest) and old C (farmland). However, the isotopic change of the atmospheric CO(2) explains only a portion of the additional 1.0 per thousand increase in delta(13)C below a depth of 20 cm. The delta(13)C of the respired CO(2) was similar to that of the organic matter in the upper soil layers but became increasingly (13)C enriched with depth, up to 2.5 per thousand relative to the organic matter. We hypothesise that this (13)C enrichment of the CO(2) as well as the residual increase in delta(13)C of the organic matter below a soil depth of 20 cm results from the increased contribution of (13)C-enriched microbially derived C with depth. Our results suggest that (13)C discrimination during microbial respiration does not contribute to the (13)C enrichment of organic matter in soils. We therefore recommend that these results should be taken into consideration when natural variations in delta(13)C of respired CO(2) are used to separate different components of soil respiration or ecosystem respiration.

Soil respiration, a key component of the global carbon cycle, is a major source of uncertainty when estimating terrestrial carbon budgets at ecosystem and higher levels. Rates of soil and root respiration are assumed to be dependent on soil temperature and soil moisture yet these factors often barely explain half the seasonal variation in soil respiration. We here found that soil moisture (range 16.5-27.6% of dry weight) and soil temperature (range 8-17.5 degrees C) together explained 55% of the variance (cross-validated explained variance; Q2) in soil respiration rate (range 1.0-3.4 micromol C m(-2) s(-1)) in a Norway spruce (Picea abies) forest. We hypothesised that this was due to that the two components of soil respiration, root respiration and decomposition, are governed by different factors. We therefore applied PLS (partial least squares regression) multivariate modelling in which we, together with below ground temperature and soil moisture, used the recent above ground air temperature and air humidity (vapour pressure deficit, VPD) conditions as x-variables. We found that air temperature and VPD data collected 1-4 days before respiration measurements explained 86% of the seasonal variation in the rate of soil respiration. The addition of soil moisture and soil temperature to the PLS-models increased the Q2 to 93%. delta13C analysis of soil respiration supported the hypotheses that there was a fast flux of photosynthates to root respiration and a dependence on recent above ground weather conditions. Taken together, our results suggest that shoot activities the preceding 1-6 days influence, to a large degree, the rate of root and soil respiration. We propose this above ground influence on soil respiration to be proportionally largest in the middle of the growing season and in situations when there is large day-to-day shifts in the above ground weather conditions. During such conditions soil temperature may not exert the major control on root respiration.

The two components of soil respiration, autotrophic respiration (from roots, mycorrhizal hyphae and associated microbes) and heterotrophic respiration (from decomposers), was separated in a root trenching experiment in a Norway spruce forest. In June 2003, cylinders (29.7 cm diameter) were inserted to 50 cm soil depth and respiration was measured both outside (control) and inside the trenched areas. The potential problems associated with the trenching treatment, increased decomposition of roots and ectomycorrhizal mycelia and changed soil moisture conditions, were handled by empirical modelling. The model was calibrated with respiration, moisture and temperature data of 2004 from the trenched plots as a training set. We estimate that over the first 5 months after the trenching, 45% of respiration from the trenched plots was an artefact of the treatment. Of this, 29% was a water difference effect and 16% resulted from root and mycelia decomposition. Autotrophic and heterotrophic respiration contributed to about 50% each of total soil respiration in the control plots averaged over the two growing seasons. We show that the potential problems with the trenching, decomposing roots and mycelia and soil moisture effects, can be handled by a modelling approach, which is an alternative to the sequential root harvesting technique.

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Summary• The mechanism behind the 13 C enrichment of fungi relative to plant materials is unclear and constrains the use of stable isotopes in studies of the carbon cycle in soils.• Here, we examined whether isotopic fractionation during respiration contributes to this pattern by comparing δ 13 C signatures of respired CO 2 , sporocarps and their associated plant materials, from 16 species of ectomycorrhizal or saprotrophic fungi collected in a Norway spruce forest.• The isotopic composition of respired CO 2 and sporocarps was positively correlated. The differences in δ 13 C between CO 2 and sporocarps were generally small, < ±1‰ in nine out of 16 species, and the average shift for all investigated species was 0.04‰. However, when fungal groups were analysed separately, three out of six species of ectomycorrhizal basidiomycetes respired 13 C-enriched CO 2 (up to 1.6‰), whereas three out of five species of polypores respired 13 C-depleted CO 2 (up to 1.7‰; P < 0.05). The CO 2 and sporocarps were always 13 C-enriched compared with wood, litter or roots.• Loss of 13 C-depleted CO 2 may have enriched some species in 13 C. However, that the CO 2 was consistently 13 C-enriched compared with plant materials implies that other processes must be found to explain the consistent 13 C-enrichment of fungal biomass compared with plant materials.