Microbial response to exudates in the rhizosphere of young beech trees (Fagus sylvatica L.) after dormancy

Plants act as an important link between atmosphere and soil: CO2 is transformed into carbohydrates by photosynthesis. These assimilates are distributed within the plant and translocated via roots into the rhizosphere and soil microorganisms. In this study, 3 year old European beech trees (Fagus sylvatica L.) were exposed after the chilling period to an enriched ¹³C–CO₂ atmosphere (δ¹³C = 60‰ – 80‰) at the time point when leaves development started. Temporal dynamics of assimilated carbon distribution in different plant parts, as well as into dissolved organic carbon and microbial communities in the rhizosphere and bulk soil have been investigated for a 20 days period. Photosynthetically fixed carbon could be traced into plant tissue, dissolved organic carbon and total microbial biomass, where it was utilized by different microbial communities. Due to carbon allocation into the rhizosphere, nutrient stress decreased; exudates were preferentially used by Gram-negative bacteria and (mycorrhizal) fungi, resulting in an enhanced growth. Other microorganisms, like Gram-positive bacteria and mainly micro eucaryotes benefited from the exudates via food web development. Overall our results indicate a fast turnover of exudates and the development of initial food web structures. Additionally a transport of assimilated carbon into bulk soil by (mycrorhizal) fungi was observed.     

Three-source partitioning of CO2 efflux from soil planted with maize by C natural abundance fails due to inactive microbial biomass

A theoretical approach to the partitioning of carbon dioxide (CO₂) efflux from soil with a C₃ vegetation history planted with maize (Zea mays), a C₄ plant, into three sources, root respiration (RR), rhizomicrobial respiration (RMR), and microbial soil organic matter (SOM) decomposition (SOMD), was examined. The δ¹³C values of SOM, roots, microbial biomass, and total CO₂ efflux were measured during a 40-day growing period. A three-source isotopic mass balance based on the measured δ¹³C values and on assumptions made in other studies showed that RR, RMR, and SOMD amounted to 91%, 4%, and 5%, respectively. Two assumptions were thoroughly examined in a sensitivity analysis: the absence of ¹³C fractionation and the conformity of δ¹³C of microbial CO₂ and that of microbial biomass. This approach strongly overestimated RR and underestimated RMR and microbial SOMD. CO₂ efflux from unplanted soil was enriched in ¹³C by 2.0‰ compared to microbial biomass. The consideration of this ¹³C fractionation in the mass balance equation changed the proportions of RR and RMR by only 4% and did not affect SOMD. A calculated δ¹³C value of microbial CO₂ by a mass balance equation including active and inactive parts of microbial biomass was used to adjust a hypothetical below-ground CO₂ partitioning to the measured and literature data. The active microbial biomass in the rhizosphere amounted to 37% to achieve an appropriate ratio between RR and RMR compared to measured data. Therefore, the three-source partitioning approach failed due to a low active portion of microbial biomass, which is the main microbial CO₂ source controlling the δ¹³C value of total microbial biomass. Since fumigation-extraction reflects total microbial biomass, its δ¹³C value was unsuitable to predict δ¹³C of released microbial CO₂ after a C₃-C₄ vegetation change. The second adjustment to the CO₂ partitioning results in the literature showed that at least 71% of the active microbial biomass utilizing maize rhizodeposits would be necessary to achieve that proportion between RR and RMR observed by other approaches based on ¹⁴C labelling. The method for partitioning total below-ground CO₂ efflux into three sources using a natural ¹³C labelling technique failed due to the small proportion of active microbial biomass in the rhizosphere. This small active fraction led to a discrepancy between δ¹³C values of microbial biomass and of microbially respired CO₂.     

Compartmentalization of the soil animal food web as indicated by dual analysis of stable isotope ratios (N/N and C/C)

The soil animal food web has become a focus of recent ecological research but trophic relationships still remain enigmatic for many taxa. Analysis of stable isotope ratios of N and C provides a powerful tool for disentangling food web structure. In this study, animals, roots, soil and litter material from a temperate deciduous forest were analysed. The combined measurement of δ¹⁵N and δ¹³C provided insights into the compartmentalization of the soil animal food web. Leaf litter feeders were separated from animals relying mainly on recent belowground carbon resources and from animals feeding on older carbon. The trophic pathway of leaf litter-feeding species appears to be a dead end, presumably because leaf litter feeders (mainly diplopods and oribatid mites) are unavailable to predators due to large size and/or strong sclerotization. Endogeic earthworms that rely on older carbon also appear to exist in predator-free space. The data suggest that the largest trophic compartment constitutes of ectomycorrhizal feeders and their predators. Additionally, there is a smaller trophic compartment consisting of predators likely feeding on enchytraeids and potentially nematodes.     

Sources of CO2 efflux from soil and review of partitioning methods

Five main biogenic sources of CO₂ efflux from soils have been distinguished and described according to their turnover rates and the mean residence time of carbon. They are root respiration, rhizomicrobial respiration, decomposition of plant residues, the priming effect induced by root exudation or by addition of plant residues, and basal respiration by microbial decomposition of soil organic matter (SOM). These sources can be grouped in several combinations to summarize CO₂ efflux from the soil including: root-derived CO₂, plant-derived CO₂, SOM-derived CO₂, rhizosphere respiration, heterotrophic microbial respiration (respiration by heterotrophs), and respiration by autotrophs. These distinctions are important because without separation of SOM-derived CO₂ from plant-derived CO₂, measurements of total soil respiration have very limited value for evaluation of the soil as a source or sink of atmospheric CO₂ and for interpreting the sources of CO₂ and the fate of carbon within soils and ecosystems. Additionally, the processes linked to the five sources of CO₂ efflux from soil have various responses to environmental variables and consequently to global warming. This review describes the basic principles and assumptions of the following methods which allow SOM-derived and root-derived CO₂ efflux to be separated under laboratory and field conditions: root exclusion techniques, shading and clipping, tree girdling, regression, component integration, excised roots and insitu root respiration; continuous and pulse labeling, ¹³C natural abundance and FACE, and radiocarbon dating and bomb-¹⁴C. A short sections cover the separation of the respiration of autotrophs and that of heterotrophs, i.e. the separation of actual root respiration from microbial respiration, as well as methods allowing the amount of CO₂ evolved by decomposition of plant residues and by priming effects to be estimated. All these methods have been evaluated according to their inherent disturbance of the ecosystem and C fluxes, and their versatility under various conditions. The shortfalls of existing approaches and the need for further development and standardization of methods are highlighted.     

C fractionation at the root-microorganisms-soil interface: A review and outlook for partitioning studies

Natural variations of the ¹³C/¹²C ratio have been frequently used over the last three decades to trace C sources and fluxes between plants, microorganisms, and soil. Many of these studies have used the natural-¹³C-labelling approach, i.e. natural δ¹³C variation after C₃-C₄ vegetation changes. In this review, we focus on ¹³C fractionation in main processes at the interface between roots, microorganisms, and soil: root respiration, microbial respiration, formation of dissolved organic carbon, as well as microbial uptake and utilization of soil organic matter (SOM). Based on literature data and our own studies, we estimated that, on average, the roots of C₃ and C₄ plants are ¹³C enriched compared to shoots by +1.2 ± 0.6‰ and +0.3 ± 0.4‰, respectively. The CO₂ released by root respiration was ¹³C depleted by about −2.1 ± 2.2‰ for C₃ plants and −1.3 ± 2.4‰ for C₄ plants compared to root tissue. However, only a very few studies investigated ¹³C fractionation by root respiration. This urgently calls for further research. In soils developed under C₃ vegetation, the microbial biomass was ¹³C enriched by +1.2 ± 2.6‰ and microbial CO₂ was also ¹³C enriched by +0.7 ± 2.8‰ compared to SOM. This discrimination pattern suggests preferential utilization of ¹³C-enriched substances by microorganisms, but a respiration of lighter compounds from this fraction. The δ¹³C signature of the microbial pool is composed of metabolically active and dormant microorganisms; the respired CO₂, however, derives mainly from active organisms. This discrepancy and the preferential substrate utilization explain the δ¹³C differences between microorganisms and CO₂ by an ‘apparent’ ¹³C discrimination. Preferential consumption of easily decomposable substrates and less negative δ¹³C values were common for substances with low C/N ratios. Preferential substrate utilization was more important for C₃ soils because, in C₄ soils, microbial respiration strictly followed kinetics, i.e. microorganisms incorporated heavier C (? = +1.1‰) and respired lighter C (? = −1.1‰) than SOM. Temperature and precipitation had no significant effect on the ¹³C fractionation in these processes in C₃ soils. Increasing temperature and decreasing precipitation led, however, to increasing δ¹³C of soil C pools.Based on these ¹³C fractionations we developed a number of consequences for C partitioning studies using ¹³C natural abundance. In the framework of standard isotope mixing models, we calculated CO₂ partitioning using the natural-¹³C-labelling approach at a vegetation change from C₃ to C₄ plants assuming a root-derived fraction between 0% and 100% to total soil CO₂. Disregarding any ¹³C fractionation processes, the calculated results deviated by up to 10% from the assumed fractions. Accounting for ¹³C fractionation in the standard deviations of the C₄ source and the mixing pool did not improve the exactness of the partitioning results; rather, it doubled the standard errors of the CO₂ pools. Including ¹³C fractionations directly into the mass balance equations reproduced the assumed CO₂ partitioning exactly. At the end, we therefore give recommendations on how to consider ¹³C fractionations in research on carbon flows between plants, microorganisms, and soil.     

Microbial immobilisation of C rhizodeposits in rhizosphere and root-free soil under continuous C labelling of oats

A greenhouse experiment was conducted by growing oats (Avenasativa L.) in a continuously ¹³CO₂ labeled atmosphere. The allocation of ¹³C-labeled photosynthates in plants, microbial biomass in rhizosphere and root-free soil, pools of soil organic C, and CO₂ emissions were examined over the plant's life cycle. To isolate rhizosphere from root-free soil, plant seedlings were placed into bags made of nylon monofilament screen tissue (16 μm mesh) filled with soil. Two peaks of ¹³C in rhizosphere pools of microbial biomass and dissolved organic carbon (DOC), as well as in CO₂ emissions at the earing and ripeness stages were revealed. These ¹³C maxima corresponded to: (i) the end of rapid root growth and (ii) beginning of root decomposition, respectively. The δ¹³C values of microbial biomass were higher than those of DOC and of soil organic matter (SOM). The microbial biomass C accounted for up to 56 and 39% of ¹³C recovered in the rhizosphere and root-free soil, respectively. Between 4 and 28% of ¹³C assimilated was recovered in the root-free soil. Depending on the phenological stage, the contribution of root-derived C to total CO₂ emission from soil varied from 61 to 92% of total CO₂ evolved, including 4-23% attributed to rhizomicrobial respiration. While 81-91% of C substrates used for microbial growth in the root-free soil and rhizosphere came from SOM, the remaining 9-19% of C substrates utilized by the microbial biomass was attributable to rhizodeposition. The use of continuous isotopic labelling and physical separation of root-free and rhizosphere soil, combined with natural ¹³C abundance were effective in gaining new insight on soil and rhizosphere C-cycling.     

Seasonal dynamics of leaf- and root-derived C in arctic tundra mesocosms

We investigated contributions of leaf litter, root litter and root-derived organic material to tundra soil carbon (C) storage and transformations. ¹⁴C-labeled materials were incubated for 32 weeks in moist tussock tundra soil cores under controlled climate conditions in growth chambers, which simulated arctic fall, winter, spring and summer temperatures and photoperiods. In addition, we tested whether the presence of living plants altered litter and soil organic matter (SOM) decomposition by planting shoots of the sedge Eriophorum vaginatum in half of the cores. Our results suggest that root litter accounted for the greatest C input and storage in these tundra soils, while leaf litter was rapidly decomposed and much of the C lost to respiration. We observed transformations of ¹⁴C between fractions even when total C appeared unchanged, allowing us to elucidate sources and sinks of C used by soil microorganisms. Initial sources of C included both water soluble (WS) and acid-soluble (AS) fractions, primarily comprised of carbohydrates and cellulose, respectively. The acid-insoluble (AIS) fraction appeared to be a sink for C when conditions were favorable for plant growth. However, decreases in ¹⁴C activity from the AIS fraction between the fall and spring harvests in all treatments indicated that microorganisms consumed recalcitrant C compounds when soil temperatures were below 0 °C. In planted leaf litter cores and in both planted and unplanted SOM cores, the greatest amounts of ¹⁴C at the end of the experiment were found in the AIS fraction, suggesting a high rate of humification or accumulation of decay-resistant plant tissues. In unplanted leaf litter cores and planted and unplanted root litter cores most of the ¹⁴C remaining at the end of the experiment was in the AS fraction suggesting less extensive humification of leaf and root detritus. Overall, the presence of living plants stimulated decomposition of leaf litter by creating favorable conditions for microbial activity at the soil surface. In contrast, plants appeared to inhibit decomposition of root litter and SOM, perhaps because of microbial preferences for newer, more labile inputs from live roots.     

Effects of root and litter exclusion on soil CO2 efflux and microbial biomass in wet tropical forests

We examined the effects of root and litter exclusion on the rate of soil CO₂ efflux and microbial biomass at a soil depth of 25 cm in a secondary forest (dominated by Tabebuia heterophylla) and a pine (Pinus caribaea) plantation in the Luquillo Experimental Forest in Puerto Rico. The experimental plots were initially established in 1990, when root, forest floor mass and new litterfall were excluded for 7 y since then. Soil respiration was significantly reduced in the litter and root exclusion plots in both the secondary forest and the pine plantation compared with the control. Root exclusion had a greater effect on soil CO₂ efflux than the litter exclusion in the plantation, whereas a reversed pattern was observed in the secondary forest. The reduction of microbial biomass in the root exclusion plot was greater in the secondary forest (59%) than in the plantation (31%), while there was no difference of the reduction in the litter exclusion plots between these forests. Our results suggest that above-ground input and roots (root litter and exudates) differentially affect soil CO₂ efflux under different vegetation types.     

Sources and mechanisms of priming effect induced in two grassland soils amended with slurry and sugar

The mechanisms and specific sources of priming effects, i.e. short term changes of soil organic matter (SOM) decomposition after substance addition, are still not fully understood. These uncertainties are partly method related, i.e. until now only two C sources in released CO₂ could be identified. We used a novel approach separating three carbon (C) sources in CO₂ efflux from soil. The approach is based on combination of different substances originated from C₃ or C₄ plants in different treatments and identical transformation of substances like C₃ sugar (from sugar beet) and C₄ sugar (from sugar cane). We investigated the influence of the addition of two substances having different microbial utilizability, i.e. slurry and sugar on the SOM or/and slurry decomposition in two grassland soils with different levels of C_org (2.3 vs. 5.1% C). Application of slurry to the soil slightly accelerated the SOM decomposition. Addition of sugar lead to changes of SOM and slurry decomposition clearly characterized by two phases: immediately after sugar addition, the microorganisms switched from the decomposition of hardly utilizable SOM to the decomposition of easily utilizable sugar. This first phase was very short (2-3 days), hence was frequently missed in other experiments. The second phase showed a slightly increased slurry and SOM decomposition (compared to the soil without sugar). The separation of three sources in CO₂ efflux from grassland soils allowed us to conclude that the C will be utilized according to its utilizability: sugar>slurry>SOM. Additionally, decomposition of more inert C (here SOM) during the period of intensive sugar decomposition was strongly reduced (negative priming effect). We conclude that, priming effects involve a chain of mechanisms: (i) preferential substrate utilization, (ii) activation of microbial biomass by easily utilizable substrate (iii) subsequent increased utilization of following substrates according to their utilizability, and (iv) decline to initial state.     

Determination of the fate of C labelled maize and wheat rhizodeposit-C in two agricultural soils in a greenhouse experiment under C-CO2-enriched atmosphere

A deeper understanding of the contribution of carbon (C) released by plant roots (rhizodeposition) to soil organic matter (SOM) can help to increase our knowledge of global C-cycling. These insights can eventually lead to sustainable management of SOM especially in agricultural systems. This study was conducted to determine the fate of ¹³C labelled rhizodeposit-C of maize and wheat plants. They were grown in a greenhouse in permeable nylon bags filled with upper soil material from two agricultural soils of the same location, but with different crop yields. The bags were placed into pots, which were also filled with soil surrounding the bags. Soil inside the bags was considered as rhizosphere soil, wheras the one outside the bags represented bulk soil. The contributions of rhizodeposits to water extractable organic carbon (WEOC), microbial biomass-C (MB-C), CO₂-C evolution, and total organic carbon (C_org) were investigated during a 7-week growing period. The WEOC, MB-C, CO₂-C, C_org contents and the respective δ¹³C values were determined regularly, and a newly developed method for determining δ¹³C values in soil extracts was applied.In both soils, regardless of crop yield potential, significant incorporation of rhizodeposition-derived C was observed in the MB-C, CO₂-C, and C_org pool, but not in the WEOC. The pattern of C incorporation into the different pools was the same for both soils with both plants, and rhizodeposit-derived C was recovered in the order MB-C<C_org<CO₂-C. This showed that rhizodeposits were mainly respired, but since C_org was the second largest pool of the overall balances, they were also stabilized in the soils at least in the short term. It is suggested that the increased SOM mineralization observed in this study (positive priming effects) was probably induced by C exchange processes between the soil matrix and soluble rhizodeposits. Moreover, soluble rhizodeposit-C was detected in MB-C and CO₂-C evolved outside the direct root zone, showing the availability of these C-components in the bulk soil.     

Effluxed CO2-C from sterilized and unsterilized treatments of a calcareous soil

Soil inorganic carbon (C) represents a substantial C pool in arid ecosystems, yet little data exist on the contribution of this pool to ecosystem C fluxes. A closed jar incubation study was carried out to test the hypothesis that CO₂-¹³C production and response to sterilization would differ in a calcareous (Mojave Desert) soil and a non-calcareous (Oklahoma Prairie) soil due to contributions of carbonate-derived CO₂. In addition to non-sterilized controls, soils were subjected to sterilization treatments (unbuffered HgCl₂ addition for Oklahoma soil and unbuffered HgCl₂ addition, buffered HgCl₂ addition, and autoclaving for Mojave Desert soil) to decrease biotic respiration and more readily measure abiotic CO₂ flux. Temperature and moisture treatments were also included with sterilization treatments in a factorial design.The rate of CO₂ production in both soils was significantly decreased (36-87%) by sterilization, but sterilization treatments differed in effectiveness. Sterilization had no significant effect on effluxed CO₂-¹³C values in the non-calcareous Oklahoma Prairie soil and autoclaved Mojave Desert soil as compared to their respective non-sterilized controls. However, sterilization significantly altered CO₂-¹³C values in Mojave Desert soil HgCl₂ sterilization treatments (both buffered and non-buffered). Plots of 1/CO₂ versus CO₂-δ¹³C (similar to Keeling plots) indicated that the source CO₂-δ¹³C value of the Oklahoma Prairie soil treatments was similar to the δ¹³C value of soil organic matter [(SOM); −17.76‰ VPDB] whereas the source for the (acidic) unbuffered-HgCl₂ sterilized Mojave Desert soil was similar to the δ¹³C value of carbonates (−0.93‰ VPDB). The source CO₂-δ¹³C value of non-sterilized and autoclaved (−18.4‰ VPDB) Mojave Desert soil treatments was intermediate between SOM (−21.43‰ VPDB) and carbonates and indicates up to 13% of total C efflux may be from abiotic sources in calcareous soils.     

Repeated CO2 pulse-labelling reveals an additional net gain of soil carbon during growth of spring wheat under free air carbon dioxide enrichment (FACE)

Rising levels of atmospheric CO₂ have often been found to increase above and belowground biomass production of C3 plants. The additional translocation of organic matter into soils by increased root mass and exudates are supposed to possibly increase C pools in terrestrial ecosystems. Corresponding investigations were mostly conducted under more or less artificial indoor conditions with disturbed soils. To overcome these limitations, we conducted a ¹⁴CO₂ pulse-labelling experiment within the German FACE project to elucidate the role of an arable crop system in carbon sequestration under elevated CO₂. We cultivated spring wheat cv. “Minaret” with usual fertilisation and ample water supply in stainless steel cylinders forced into the soil of a control and a FACE plot. Between stem elongation and beginning of ripening the plants were repeatedly pulse-labelled with ¹⁴CO₂ in the field. Soil born total CO₂ and ¹⁴CO₂ was monitored daily till harvest. Thereafter, the distribution of ¹⁴C was analysed in all plant parts, soil, soil mineral fractions and soil microbial biomass. Due to the small number of grown wheat plants (40) in each ring and the inherent low statistical power, no significant above and belowground growth effect of elevated CO₂ was detected at harvest. But in comparison to ambient conditions, 28% more ¹⁴CO₂ and 12% more total CO₂ was evolved from soil under elevated CO₂ (550 μmol CO₂ mol⁻¹). In the root-free soil 27% more residual ¹⁴C was found in the FACE soil than in the soil from the ambient ring. In soil samples from both treatments about 80% of residual ¹⁴C was found in the clay fraction and 7% in the silt fraction. Very low ¹⁴C contents in the CFE extracts of microbial biomass in the soil from both CO₂ treatments did not allow assessing their influence on this parameter. Since the calculated specific radioactivity of soil born ¹⁴CO₂ gave no indication of an accelerated priming effect in the FACE soil, we conclude that wheat plants grown under elevated CO₂ can contribute to an additional net carbon gain in soils.     

Soil organic matter dynamics in grassland soils under elevated CO2: Insights from long-term incubations and stable isotopes

Elevated atmospheric carbon dioxide (CO₂) levels generally stimulate carbon (C) uptake by plants, but the fate of this additional C largely remains unknown. This uncertainty is due in part to the difficulty in detecting small changes in soil carbon pools. We conducted a series of long-term (170-330 days) laboratory incubation experiments to examine changes in soil organic matter pool sizes and turnover rates in soil collected from an open-top chamber (OTC) elevated CO₂ study in Colorado shortgrass steppe. We measured concentration and isotopic composition of respired CO₂ and applied a two-pool exponential decay model to estimate pool sizes and turnover rates of active and slow C pools. The active and slow C pools of surface soils (5-10 cm depth) were increased by elevated CO₂, but turnover rates of these pools were not consistently altered. These findings indicate a potential for C accumulation in near-surface soil C pools under elevated CO₂. Stable isotopes provided evidence that elevated CO₂ did not alter the decomposition rate of new C inputs. Temporal variations in measured δ¹³C of respired CO₂ during incubation probably resulted mainly from the decomposition of changing mixtures of fresh residue and older organic matter. Lignin decomposition may have contributed to declining δ¹³C values late in the experiments. Isotopic dynamics during decomposition should be taken into account when interpreting δ¹³C measurements of soil respiration. Our study provides new understanding of soil C dynamics under elevated CO₂ through the use of stable C isotope measurements during microbial organic matter mineralization.     

Turnover of soil organic matter and of microbial biomass under C3-C4 vegetation change: Consideration of C fractionation and preferential substrate utilization

Two processes contribute to changes of the δ¹³C signature in soil pools: ¹³C fractionation per se and preferential microbial utilization of various substrates with different δ¹³C signature. These two processes were disentangled by simultaneously tracking δ¹³C in three pools - soil organic matter (SOM), microbial biomass, dissolved organic carbon (DOC) - and in CO₂ efflux during incubation of 1) soil after C₃-C₄ vegetation change, and 2) the reference C₃ soil.The study was done on the Ap horizon of a loamy Gleyic Cambisol developed under C₃ vegetation. Miscanthus giganteus - a perennial C₄ plant - was grown for 12 years, and the δ¹³C signature was used to distinguish between ‘old’ SOM (>12 years) and ‘recent’ Miscanthus-derived C (<12 years). The differences in δ¹³C signature of the three C pools and of CO₂ in the reference C₃ soil were less than 1‰, and only δ¹³C of microbial biomass was significantly different compared to other pools. Nontheless, the neglecting of isotopic fractionation can cause up to 10% of errors in calculations. In contrast to the reference soil, the δ¹³C of all pools in the soil after C₃-C₄ vegetation change was significantly different. Old C contributed only 20% to the microbial biomass but 60% to CO₂. This indicates that most of the old C was decomposed by microorganisms catabolically, without being utilized for growth. Based on δ¹³C changes in DOC, CO₂ and microbial biomass during 54 days of incubation in Miscanthus and reference soils, we concluded that the main process contributing to changes of the δ¹³C signature in soil pools was preferential utilization of recent versus old C (causing an up to 9.1‰ shift in δ¹³C values) and not ¹³C fractionation per se.Based on the δ¹³C changes in SOM, we showed that the estimated turnover time of old SOM increased by two years per year in 9 years after the vegetation change. The relative increase in the turnover rate of recent microbial C was 3 times faster than that of old C indicating preferential utilization of available recent C versus the old C.Combining long-term field observations with soil incubation reveals that the turnover time of C in microbial biomass was 200 times faster than in total SOM. Our study clearly showed that estimating the residence time of easily degradable microbial compounds and biomarkers should be done at time scales reflecting microbial turnover times (days) and not those of bulk SOM turnover (years and decades). This is necessary because the absence of C reutilization is a prerequisite for correct estimation of SOM turnover. We conclude that comparing the δ¹³C signature of linked pools helps calculate the relative turnover of old and recent pools.     

Priming effects: Interactions between living and dead organic matter

In this re-evaluation of our 10-year old paper on priming effects, I have considered the latest studies and tried to identify the most important needs for future research. Recent publications have shown that the increase or decrease in soil organic matter mineralization (measured as changes of CO₂ efflux and N mineralization) actually results from interactions between living (microbial biomass) and dead organic matter. The priming effect (PE) is not an artifact of incubation studies, as sometimes supposed, but is a natural process sequence in the rhizosphere and detritusphere that is induced by pulses or continuous inputs of fresh organics. The intensity of turnover processes in such hotspots is at least one order of magnitude higher than in the bulk soil. Various prerequisites for high-quality, informative PE studies are outlined: calculating the budget of labeled and total C; investigating the dynamics of released CO₂ and its sources; linking C and N dynamics with microbial biomass changes and enzyme activities; evaluating apparent and real PEs; and assessing PE sources as related to soil organic matter stabilization mechanisms. Different approaches for identifying priming, based on the assessment of more than two C sources in CO₂ and microbial biomass, are proposed and methodological and statistical uncertainties in PE estimation and approaches to eliminating them are discussed. Future studies should evaluate directions and magnitude of PEs according to expected climate and land-use changes and the increased rhizodeposition under elevated CO₂ as well as clarifying the ecological significance of PEs in natural and agricultural ecosystems. The conclusion is that PEs - the interactions between living and dead organic matter - should be incorporated in models of C and N dynamics, and that microbial biomass should regarded not only as a C pool but also as an active driver of C and N turnover.     

Moisture modulates rhizosphere effects on C decomposition in two different soil types

While it is well known that soil moisture directly affects microbial activity and soil organic matter (SOM) decomposition, it is unclear if the presence of plants alters these effects through rhizosphere processes. We studied soil moisture effects on SOM decomposition with and without sunflower and soybean. Plants were grown in two different soil types with soil moisture contents of 45% and 85% of field capacity in a greenhouse experiment. We continuously labeled plants with depleted ¹³C, which allowed us to separate plant-derived CO₂-C from original soil-derived CO₂-C in soil respiration measurements. We observed an overall increase in soil-derived CO₂-C efflux in the presence of plants (priming effect) in both soils. On average a greater priming effect was found in the high soil moisture treatment (up to 76% increase in soil-derived CO₂-C compared to control) than in the low soil moisture treatment (up to 52% increase). Greater plant-derived CO₂-C and plant biomass in the high soil moisture treatment contributed to greater priming effects, but priming effects remained significantly higher in the high moisture treatment than in the low moisture treatment after correcting for the effects of plant-derived CO₂-C and plant biomass. The response to soil moisture particularly occurred in the sandy loam soil by the end of the experiment. Possibly, production of root exudates increased with increased soil moisture content. Root exudation of labile C may also have become more effective in stimulating microbial decomposition in the higher soil moisture treatment and sandy loam soil. Our results indicate that moisture conditions significantly modulate rhizosphere effects on SOM decomposition.     

Black carbon decomposition and incorporation into soil microbial biomass estimated by C labeling

Incomplete combustion of organics such as vegetation or fossil fuel led to accumulation of charred products in the upper soil horizon. Such charred products, frequently called pyrogenic carbon or black carbon (BC), may act as an important long-term carbon (C) sink because its microbial decomposition and chemical transformation is probably very slow. Direct estimations of BC decomposition rates are absent because the BC content changes are too small for any relevant experimental period. Estimations based on CO₂ efflux are also unsuitable because the contribution of BC to CO₂ is too small compared to soil organic matter (SOM) and other sources.We produced BC by charring ¹⁴C labeled residues of perennial ryegrass (Lolium perenne). We then incubated this ¹⁴C labeled BC in Ah of a Haplic Luvisol soil originated from loess or in loess for 3.2 years. The decomposition rates of BC were estimated based on ¹⁴CO₂ sampled 44 times during the 3.2 years incubation period (1181 days). Additionally we introduced five repeated treatments with either 1) addition of glucose as an energy source for microorganisms to initiate cometabolic BC decomposition or 2) intensive mixing of the soil to check the effect of mechanical disturbance of aggregates on BC decomposition. Black carbon addition amounting to 20% of C_org of the soil or 200% of C_org of loess did not change total CO₂ efflux from the soil and slightly decreased it from the loess. This shows a very low BC contribution to recent CO₂ fluxes. The decomposition rates of BC calculated based on ¹⁴C in CO₂ were similar in soil and in loess and amounted to 1.36 10⁻⁵ d⁻¹ (=1.36 10⁻³% d⁻¹). This corresponds to a decomposition of about 0.5% BC per year under optimal conditions. Considering about 10 times slower decomposition of BC under natural conditions, the mean residence time (MRT) of BC is about 2000 years, and the half-life is about 1400 years. Considering the short duration of the incubation and the typical decreasing decomposition rates with time, we conclude that the MRT of BC in soils is in the range of millennia.The strong increase in BC decomposition rates (up to 6 times) after adding glucose and the decrease of this stimulation after 2 weeks in the soil (and after 3 months in loess) allowed us to conclude cometabolic BC decomposition. This was supported by higher stimulation of BC decomposition by glucose addition compared to mechanical disturbance as well as higher glucose effects in loess compared to the soil. The effect of mechanical disturbance was over within 2 weeks. The incorporation of BC into microorganisms (fumigation/extraction) after 624 days of incubation amounted to 2.6 and 1.5% of ¹⁴C input into soil and loess, respectively. The amount of BC in dissolved organic carbon (DOC) was below the detection limit (<0.01%) showing no BC decomposition products in water leached from the soil.We conclude that applying ¹⁴C labeled BC opens new ways for very sensitive tracing of BC transformation products in released CO₂, microbial biomass, DOC, and SOM pools with various properties.     

Rhizodeposition-induced decomposition increases N availability to wild and cultivated wheat genotypes under elevated CO2

Elevated CO₂ may increase nutrient availability in the rhizosphere by stimulating N release from recalcitrant soil organic matter (SOM) pools through enhanced rhizodeposition. We aimed to elucidate how CO₂-induced increases in rhizodeposition affect N release from recalcitrant SOM, and how wild versus cultivated genotypes of wheat mediated differential responses in soil N cycling under elevated CO₂. To quantify root-derived soil carbon (C) input and release of N from stable SOM pools, plants were grown for 1 month in microcosms, exposed to ¹³C labeling at ambient (392 μmol mol⁻¹) and elevated (792 μmol mol⁻¹) CO₂ concentrations, in soil containing ¹⁵N predominantly incorporated into recalcitrant SOM pools. Decomposition of stable soil C increased by 43%, root-derived soil C increased by 59%, and microbial-¹³C was enhanced by 50% under elevated compared to ambient CO₂. Concurrently, plant ¹⁵N uptake increased (+7%) under elevated CO₂ while ¹⁵N contents in the microbial biomass and mineral N pool decreased. Wild genotypes allocated more C to their roots, while cultivated genotypes allocated more C to their shoots under ambient and elevated CO₂. This led to increased stable C decomposition, but not to increased N acquisition for the wild genotypes. Data suggest that increased rhizodeposition under elevated CO₂ can stimulate mineralization of N from recalcitrant SOM pools and that contrasting C allocation patterns cannot fully explain plant mediated differential responses in soil N cycling to elevated CO₂.     

Carbon dynamics determined by natural C abundance in microcosm experiments with soils from long-term maize and rye monocultures

Understanding carbon dynamics in soil is the key to managing soil organic matter. Our objective was to quantify the carbon dynamics in microcosm experiments with soils from long-term rye and maize monocultures using natural ¹³C abundance. Microcosms with undisturbed soil columns from the surface soil (0-25 cm) and subsoil (25-50 cm) of plots cultivated with rye (C₃-plant) since 1878 and maize (C₄-plant) since 1961 with and without NPK fertilization from the long-term experiment ‘Ewiger Roggen’ in Halle, Germany, were incubated for 230 days at 8 °C and irrigated with 2 mm 10⁻² M CaCl₂ per day. Younger, C₄-derived and older, C₃-derived percentages of soil organic carbon (SOC), dissolved organic carbon (DOC), microbial biomass (C_mic) and CO₂ from heterothropic respiration were determined by natural ¹³C abundance. The percentage of maize-derived carbon was highest in CO₂ (42-79%), followed by C_mic (23-46%), DOC (5-30%) and SOC (5-14%) in the surface soils and subsoils of the maize plots. The percentage of maize-derived C was higher for the NPK plot than for the unfertilized plot and higher for the surface soils than for the subsoils. Specific production rates of DOC, CO₂-C and C_mic from the maize-derived SOC were 0.06-0.08% for DOC, 1.6-2.6% for CO₂-C and 1.9-2.7% for C_mic, respectively, and specific production rates from rye-derived SOC of the continuous maize plot were 0.03-0.05% for DOC, 0.1-0.2% for CO₂-C and 0.3-0.5% for C_mic. NPK fertilization did not affect the specific production rates. Strong correlations were found between C₄-derived C_mic and C₄-derived SOC, DOC and CO₂-C (r≥0.90), whereas the relationship between C₃-derived C_mic and C₃-derived SOC, DOC and CO₂-C was not as pronounced (r≤0.67). The results stress the different importance of former (older than 40 years) and recent (younger than 40 years) litter C inputs for the formation of different C pools in the soil.     

Root-derived carbon in soil respiration and microbial biomass determined by 14C and 13C

Two approaches to quantitatively estimating root-derived carbon in soil CO₂ efflux and in microbial biomass were compared under controlled conditions. In the ¹⁴C labelling approach, maize (Zea mays) was pulse labelled and the tracer was chased in plant and soil compartments. Root-derived carbon in CO₂ efflux and in microbial biomass was estimated based on a linear relationship between the plant shoots and the below-ground compartment. Since the maize plants were grown on C₃ soil, in a second approach the differences in ¹³C natural abundance between C₃ and C₄ plants were used to calculate root-derived carbon in the CO₂ efflux and in the microbial biomass. The root-derived carbon in the total CO₂ efflux was between 69% and 94% using the ¹⁴C labelling approach and between 86% and 94% in the natural ¹³C labelling approach. At a ¹³C fractionation measured to be 5.2‰ between soil organic matter (SOM) and CO₂, the root-derived contribution to CO₂ ranged from 70% to 88% and was much closer to the results of the ¹⁴C labelling approach. Root-derived contributions to the microbial biomass carbon ranged from 2% to 9% using ¹⁴C labelling and from 16% to 36% using natural ¹³C labelling. At a 3.2‰ ¹³C fractionation between SOM and microbial biomass, both labelling approaches yielded an equal contribution of root-derived C in the microbial biomass. Both approaches may therefore be used to partition CO₂ efflux and to quantify the C sources of microbial biomass. However, the assumed ¹³C fractionation strongly affects the contributions of individual C sources.