Qualitative assessment of rhizodeposits in non‐sterile soil by analytical pyrolysis

Estimation of the amount of root exudates and simultaneous identification of their composition in non‐sterile soil is a challenging objective in rhizosphere research. We coupled 3 methods: (1) labeling of corn in ¹⁴CO₂ atmosphere to separate root‐derived and soil‐derived organic substances in the rhizosphere, (2) a previously developed leaching method to collect rhizodeposits, and (3) pyrolysis field ionization mass spectrometry (Py‐FIMS) to investigate the molecular‐chemical composition of rhizodeposits. Eluted rhizodeposits accounted for 2.8 % (Loam) and 0.97 % (nutrient solution in quartz sand) of recovered ¹⁴C and showed clear differences in composition between the growth substrates. The ¹⁴CO₂ evolved mostly by root respiration accounted for 3.5–4.0 % without significant differences according to growth substrate or diurnal dynamics. Principal component analysis of the Py‐FI mass spectra of leachates showed a clear diurnal dynamics of the amount and the composition of corn rhizodeposits collected during day‐time and night‐time. Differences originated mostly from signals assigned to carbohydrates, sterols, and peptides. This approach is recommended for forthcoming studies of rhizodeposition in different soil substrates, crops grown, and time‐series of exudate sampling.     

Qualitative and quantitative analysis of water‐soluble root exudates in relation to plant species and development

The composition of root‐derived substances is of great importance for the understanding of processes in the rhizosphere. Therefore, methods allowing a comprehensive collection and chemical analysis of the organic root exudates are necessary. In this study, we compare different methods with regard to their suitability to collect and characterize root exudates. Because the percolation or water logging method failed to quantitatively extract root exudates, a dipping method was developed which allowed an almost complete sampling of coldwater‐soluble root exudates. By ¹⁴CO₂ labeling of the shoots the composition of root exudates was found to be influenced by plant species and growth stage. In comparison to pea plants maize plants had a higher share of carboxylic acids and a lower share of sugars. Younger maize plants exuded considerably higher amounts of ¹⁴C labeled organic substances per g root dry matter than older ones. During plant development the relative amount of sugars decreased at the expense of carboxylic acids. The described methods are well suited for the elucidation of the influence of growth factors on root exudation.     

Spatially tracking carbon through the root–rhizosphere–soil system using laser ablation‐IRMS

The intimate relationships between plant roots, rhizosphere, and soil are fostered by the release of organic compounds from the plant into soil through various forms of rhizodeposition and the simultaneous harvesting of nutrients from the soil to the plant. Here we present a method to spatially track and map the migration of plant‐derived carbon (C) through roots into the rhizosphere and surrounding soil using laser ablation‐isotope ratio mass spectrometry (LA‐IRMS). We used switchgrass microcosms containing soil from field plots at the Kellogg Biological Station (Hickory Corners, Michigan, USA) which have been cropped with switchgrass since 2008. We used a ¹³CO₂ tracer to isotopically label switchgrass plants for two diel cycles and tracked subsequent movement of labeled C using the spatially specific (< 100 µm resolution) δ¹³C analysis enabled by LA‐IRMS. This approach permitted assessment of variable C flow through different roots and enabled mapping of spatial variability of C allocation to the rhizosphere. Highly ¹³C‐enriched C (consistent with production during the ¹³CO₂ application period) extended ≈ 0.5–1 mm from the root into the soil, suggesting that the majority of recent plant‐derived C was within this distance of the root after 48 h. Tracking the physical extent of root exudation into the rhizosphere can help evaluate the localization of plant‐microbe interactions in highly variable subsurface environments, and the use of the isotopic label can differentiate freshly fixed C (presumably from root exudates) from other types of subsurface C (e.g., plant necromass and microbial turnover). The LA‐IRMS technique may also serve as a valuable screening technique to identify areas of high activity for additional microbial or geochemical assays.     

Contribution of rhizomicrobial and root respiration to the CO2 emission from soil (A review)

Separate determination of root respiration and rhizomicrobial respiration is one of the most interesting, important, and methodologically complicated problems in the study of the carbon budget in soils and the subdivision of the CO₂ emission from soils into separate fluxes. In this review, we compare the main principles, the advantages and disadvantages, and the results obtained by the methods of component integration, substrate-induced respiration, respiratory capacity, girdling, isotope dilution, model rhizodeposition, modeling of the ¹⁴CO₂ efflux dynamics, exudates elution, and the δ¹³C measurements of the microbial biomass and CO₂. Summarizing the results of the determinations performed by these methods, we argue that about 40% of the rhizosphere CO₂ efflux is due to root respiration and about 60% of this efflux is due to the respiration of microorganisms decomposing root exudates.     

Root and rhizomicrobial respiration: A review of approaches to estimate respiration by autotrophic and heterotrophic organisms in soil

Partitioning the root‐derived CO₂ efflux from soil (frequently termed rhizosphere respiration) into actual root respiration (RR, respiration by autotrophs) and rhizomicrobial respiration (RMR, respiration by heterotrophs) is crucial in determining the carbon (C) and energy balance of plants and soils. It is also essential in quantifying C sources for rhizosphere microorganisms and in estimation of the C contributing to turnover of soil organic matter (SOM), as well as in linking net ecosystem production (NEP) and net ecosystem exchange (NEE). Artificial‐environment studies such as hydroponics or sterile soils yield unrealistic C‐partitioning values and are unsuitable for predicting C flows under natural conditions. To date, several methods have been suggested to separate RR and RMR in nonsterile soils: 1) component integration, 2) substrate‐induced respiration, 3) respiration by excised roots, 4) comparison of root‐derived ¹⁴CO₂ with rhizomicrobial ¹⁴CO₂ after continuous labeling, 5) isotope dilution, 6) model‐rhizodeposition technique, 7) modeling of ¹⁴CO₂ efflux dynamics, 8) exudate elution, and 9) δ¹³C of CO₂ and microbial biomass. This review describes the basic principles and assumptions of these methods and compares the results obtained in the original papers and in studies designed to compare the methods. The component‐integration method leads to strong disturbance and non‐proportional increase of CO₂ efflux from different sources. Four of the methods (5 to 8) are based on the pulse labeling of shoots in a ¹⁴CO₂ atmosphere and subsequent monitoring of ¹⁴CO₂ efflux from the soil. The model‐rhizodeposition technique and exudate‐elution procedure strongly overestimate RR and underestimate RMR. Despite alternative assumptions, isotope dilution and modeling of ¹⁴CO₂‐efflux dynamics yield similar results. In crops and grasses (wheat, ryegrass, barley, buckwheat, maize, meadow fescue, prairie grasses), RR amounts on average to 48±5% and RMR to 52±5% of root‐derived CO₂. The method based on the ¹³C isotopic signature of CO₂ and microbial biomass is the most promising approach, especially when the plants are continuously labeled in ¹³CO₂ or ¹⁴CO₂ atmosphere. The “difference” methods, i.e., trenching, tree girdling, root‐exclusion techniques, etc., are not suitable for separating the respiration by autotrophic and heterotrophic organisms because the difference methods neglect the importance of microbial respiration of rhizodeposits.     

Plant rhizodeposition — an important source for carbon turnover in soils

The soil organic matter plays a key role in ecological soil functions, and has to be considered as an important CO₂ sink on a global scale. Apart from crop residues (shoots and roots), left over on the field after harvest, carbon and nitrogen compounds are also released by plant roots into the soil during vegetation, and undergo several transformation processes. Up to now the knowledge about amount, composition, and turnover of these root‐borne compounds is still very limited. So far it could be demonstrated with different plant species, that up to 20 % of photosynthetically fixed C are released into the soil during vegetation period. These C amounts are ecological relevant. Depending on assimilate sink strength during ontogenesis, the C release varies with plant age. A large percentage of these root‐borne substances were rapidly respired by microorganisms (64—86 %). About 2—5 % of net C assimilation was kept in soil. The root exudates of maize were mainly water‐soluble (79 %), and in this fraction about 64 % carbohydrates, 22 % amino acids/amides and 14 % organic acids could be identified. Plant species and in some cases also plant cultivars varied strongly in their root exudation pattern. Under non‐sterile conditions the exuded compounds were rapidly stabilized in water‐insoluble forms and bound preferably to the soil clay fraction. The binding of root exudates to soil particles also improved soil structure by increasing aggregate stability. Future research should focus on quantification and characterization of root‐borne C compounds during the whole plant ontogenesis. Apart from pot experiments with ¹⁴CO₂ labeling, it is necessary to conduct model field experiments with ¹³CO₂ labeling in order to be able to distinguish between CO₂ originating from the soil C pool and rhizosphere respiration, originating from plant assimilates. Such a separation is necessary to assess if soils are sources or sinks of CO₂. The incorporation of root‐borne C (¹⁴C, ¹³C) into soil organic matter of different stability is also of particular interest.     

Carbon turnover in the rhizosphere under continuous plant labeling with <Superscript>13</Superscript>CO<Subscript>2</Subscript>: Partitioning of root,microbial, and rhizomicrobial respiration

The input of labeled C into the pool of soil organic matter, the CO₂ fluxes from the soil, and the contribution of root and microbial respiration to the CO₂ emission were studied in a greenhouse experiment with continuous labeling of oat plants with ¹³CO₂ using the method of the natural ¹³C abundance in the air. The carbon of the microbial biomass composed 56 and 39% of the total amounts of ¹³C photoassimilates in the rhizosphere and in the bulk soil, respectively. The contribution of root respiration to the CO₂ emission from the soil reached 61–92%, including 4–23% of the rhizomicrobial respiration. The contribution of the microbial respiration to the total CO₂ emission from the soil varied from 8 to 39%. The soil organic matter served as the major carbon-containing substrate for microorganisms in the bulk soil and in the rhizosphere: 81–91% of the total amount of carbon involved in the microbial metabolism was derived from the soil organic matter.     

Contribution of exudates,arbuscular mycorrhizal fungi and litter depositions to the rhizosphere priming effect induced by grassland species

The presence of plants induces strong accelerations in soil organic matter (SOM) mineralization by stimulating soil microbial activity – a phenomenon known as the rhizosphere priming effect (RPE). The RPE could be induced by several mechanisms including root exudates, arbuscular mycorrhizal fungi (AMF) and root litter. However the contribution of each of these to rhizosphere priming is unknown due to the complexity involved in studying rhizospheric processes. In order to determine the role of each of these mechanisms, we incubated soils enclosed in nylon meshes that were permeable to exudates, or exudates & AMF or exudates, AMF and roots under three grassland plant species grown on sand. Plants were continuously labeled with ¹³C depleted CO₂ that allowed distinguishing plant-derived CO₂ from soil-derived CO₂. We show that root exudation was the main way by which plants induced RPE (58–96% of total RPE) followed by root litter. AMF did not contribute to rhizosphere priming under the two species that were significantly colonized by them i.e. Poa trivialis and Trifolium repens. Root exudates and root litter differed with respect to their mechanism of inducing RPE. Exudates induced RPE without increasing microbial biomass whereas root litter increased microbial biomass and raised the RPE mediating saprophytic fungi. The RPE efficiency (RPE/unit plant-C assimilated into microbes) was 3–7 times higher for exudates than for root litter. This efficiency of exudates is explained by a microbial allocation of fresh carbon to mineralization activity rather than to growth. These results suggest that root exudation is the main way by which plants stimulated mineralization of soil organic matter. Moreover, the plants through their exudates not only provide energy to soil microorganisms but also seem to control the way the energy is used in order to maximize soil organic matter mineralization and drive their own nutrient supply.     

Three‐source partitioning of CO2 efflux from maize field soil by 13C natural abundance

A natural‐¹³C‐labeling approach—formerly observed under controlled conditions—was tested in the field to partition total soil CO₂ efflux into root respiration, rhizomicrobial respiration, and soil organic matter (SOM) decomposition. Different results were expected in the field due to different climate, site, and microbial properties in contrast to the laboratory. Within this isotopic method, maize was planted on soil with C₃‐vegetation history and the total CO₂ efflux from soil was subdivided by isotopic mass balance. The C₄‐derived C in soil microbial biomass was also determined. Additionally, in a root‐exclusion approach, root‐ and SOM‐derived CO₂ were determined by the total CO₂ effluxes from maize (Zea mays L.) and bare‐fallow plots. In both approaches, maize‐derived CO₂ contributed 22% to 35% to the total CO₂ efflux during the growth period, which was comparable to other field studies. In our laboratory study, this CO₂ fraction was tripled due to different climate, soil, and sampling conditions. In the natural‐¹³C‐labeling approach, rhizomicrobial respiration was low compared to other studies, which was related to a low amount of C₄‐derived microbial biomass. At the end of the growth period, however, 64% root respiration and 36% rhizomicrobial respiration in relation to total root‐derived CO₂ were calculated when considering high isotopic fractionations between SOM, microbial biomass, and CO₂. This relationship was closer to the 50% : 50% partitioning described in the literature than without fractionation (23% root respiration, 77% rhizomicrobial respiration). Fractionation processes of ¹³C must be taken into account when calculating CO₂ partitioning in soil. Both methods—natural ¹³C labeling and root exclusion—showed the same partitioning results when ¹³C isotopic fractionation during microbial respiration was considered and may therefore be used to separate plant‐ and SOM‐derived CO₂ sources.     

Carbon allocation in grassland communities under drought stress followed by 14C pulse labeling

Although extreme climatic events such as drought have important consequences for belowground carbon (C) cycling, their impact on the plant-soil system of mixed plant communities is poorly understood. Our objective was to study the effect of drought on C allocation and rhizosphere-mediated CO₂ fluxes under three plant species: Lolium perenne, Festuca arundinacea and Medicago sativa grown in monocultures or mixture. The conceptual approach included ¹⁴CO₂ pulse labeling of plants grown under drought and optimum water conditions in order to be able to follow above- and belowground C allocation. After ¹⁴C pulse labeling, we traced ¹⁴C allocation to shoots and roots, soil and rhizospheric CO₂, dissolved organic carbon (DOC) and microbial biomass.Drought and plant community composition significantly affected assimilate allocation in the plant-soil system. Drought conditions changed the source sink relationship of monocultures, which transferred a relatively larger portion of assimilates to their roots compared to water sufficient plants. In contrast, plant mixture showed an increase in ¹⁴C allocation to shoots when exposed to drought.Under drought stress, root respiration was reduced for all monocultures except under the legume species. Microbial respiration remained similar in all cases showing that microbial activity was less affected by drought than root activity. This may be explained by strongly increased assimilate allocation to easily available exudates or rhizodeposits under drought. In conclusion, plant community composition may modify the impact of climatic changes on carbon allocation and belowground carbon fluxes. The presence of legume species attenuates drought effects on rhizosphere processes.     

Separation of root and microbial respiration: Comparison of three methods

In a laboratory experiment, the following methods of separating the soil CO₂ flux into the root respiration and the respiration of the rhizosphere and nonrhizosphere microorganisms were compared: (1) root exclusion, (2) component integration, and (3) ¹⁴C pulse labeling. Depending on the method used, the combined contribution of the rhizosphere microorganisms and roots varied from 18 to 40% of the total CO₂ emission; the contribution of the roots alone was 8–19%, and that of the nonrhizosphere microorganisms was 51–82%. The nonisotope methods (1 and 2) gave similar results of the separation. The pulse labeling of plants satisfactorily separated the root and microbial respiration, but it is unsuitable for determining the respiration of the nonrhizosphere microorganisms. Advantages and disadvantages of each method are discussed.     

Cattle grazing drives nitrogen and carbon cycling in a temperate salt marsh

We examined the impact of long-term cattle grazing on soil processes and microbial activity in a temperate salt marsh. Soil conditions, microbial biomass and respiration, mineralization and denitrification rates were measured in upper salt marsh that had been ungrazed or cattle grazed for several decades. Increased microbial biomass and soil respiration were observed in grazed marsh, most likely stimulated by enhanced rates of root turnover and root exudation. We found a significant positive effect of grazing on potential N mineralization rates measured in the laboratory, but this difference did not translate to in situ net mineralization measured monthly from May to September. Rates of denitrification were lowest in the grazed marsh and appeared to be limited by nitrate availability, possibly due to more anoxic conditions and lower rates of nitrification. The major effect of grazing on N cycling therefore appeared to be in limiting losses of N through denitrification, which may lead to enhanced nutrient availability to saltmarsh plants, but a reduced ability of the marsh to act as a buffer for land-derived nutrients to adjacent coastal areas. Additionally, we investigated if grazing influences the rates of turnover of labile and refractory C in saltmarsh soils by adding ¹⁴C-labelled leaf litter or root exudates to soil samples and monitoring the evolution of ¹⁴CO₂. Grazing had little effect on the rates of mineralization of ¹⁴C used as a respiratory substrate, but a larger proportion of ¹⁴C was partitioned into microbial biomass and immobilized in long- and medium-term storage pools in the grazed treatment. Grazing slowed down the turnover of the microbial biomass, which resulted in longer turnover times for both leaf litter and root exudates. Grazing may therefore affect the longevity of C in the soil and alter C storage and utilization pathways in the microbial community.     

Interaction between rhizosphere microorganisms and plant roots: 13C fluxes in the rhizosphere after pulse labeling

The input dynamics of labeled C into pools of soil organic matter and CO₂ fluxes from soil were studied in a pot experiment with the pulse labeling of oats and corn under a ¹³CO₂ atmosphere, and the contribution of the root and microbial respiration to the emission of CO₂ from the soil was determined from the fluxes of labeled C in the microbial biomass and the evolved carbon dioxide. A considerable amount of ¹³C (up to 96% of the total amount of the label found in the rhizosphere soil) was incorporated into the biomass of the rhizosphere microorganisms. The diurnal fluctuations of the labeled C pools in the microbial biomass, dissolved organic carbon, and CO₂ released in the rhizosphere of oats and corn were related to the day/night changes, i.e., to the on and off periods of the photosynthetic activity of the plants. The average contribution of the corn root respiration (70% of the total CO₂ emission from the soil surface) was higher than that of the oats roots (44%), which was related to the lower incorporation of rhizodeposit carbon into the microbial biomass in the soil under the corn plants than in the soil under the oats plants.     

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.     

Characterisation of the dynamics of C-partitioning within Lolium perenne and to the rhizosphere microbial biomass using 14C pulse chase

The dynamics of C partitioning with Lolium perenne and its associated rhizosphere was investigated in plant-soil microcosms using ¹⁴C pulse-chase labelling. The ¹⁴CO₂ pulse was introduced into the shoot chamber and the plants allowed to assimilate the label for a fixed period. The microcosm design facilitated independent monitoring of shoot and root/soil respiration during the chase period. Partitioning between above- and below-ground pools was determined between 30 min and 168 h after the pulse, and the distribution was found to vary with the length of the chase period. Initially (30 min after the pulse), the ¹⁴C was predominantly (99%) in the shoot biomass and declined thereafter. The results indicate that translocation of recent photoassimilate is rapid, with ¹⁴C detected below ground within 30 min of pulse application. The translocation rate of ¹⁴C below ground was maximal (6.2% h^-1) between 30 min and 3 h after the pulse, with greatest incorporation into the microbial biomass detected at 3 h. After 3 h, the microbial biomass ¹⁴C pool accounted for 74% of the total ¹⁴C rhizosphere pool. By 24 h, approximately 30% of ¹⁴C assimilate had been translocated below ground; thereafter ¹⁴C translocation was greatly reduced. Partitioning of recent assimilate changed with increasing CO₂ concentration. The proportion of ¹⁴C translocated below ground almost doubled from 17.76% at the ambient atmospheric CO₂ concentration (450 ppm) to 33.73% at 750 ppm CO₂ concentration. More specifically, these changes occurred in the root biomass and the total rhizosphere pools, with two- and threefold ¹⁴C increases at an elevated CO₂ concentration compared to ambient, respectively. The pulselabelling strategy developed in this study provided sufficient sensitivity to determine perturbations in C dynamics in L. perenne, in particular rhizosphere C pools, in response to an elevated atmospheric CO₂ concentration.     

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.     

Role of arbuscular mycorrhizal network in carbon and phosphorus transfer between plants

Intercropping with aerobic rice or arbuscular mycorrhizal fungi (AMF) colonization alleviated watermelon wilt disease, which is likely attributed to rice root exudates or AMF depressing watermelon wilt pathogen. However, it is unclear whether rice root exudates transfers to watermelon rhizosphere soil and whether AMF affects the transfer of rice root exudates to watermelon rhizosphere soil. A rhizobox experiment, with aerobic rice under ¹⁴?CO₂, was conducted to investigate the effect of AMF colonization on carbon (C) transfer from rice to watermelon and on phosphorus (P) uptake by both watermelon and rice. The rhizobox was separated into labelling side (L side) and sampling side (S side) by inserting nylon mesh in the middle of the box. The L side was planted with aerobic rice, and the S side was aerobic rice (monocropping) or watermelon (intercropping). When ¹⁴?CO₂ was added to rice canopy at the L side, ¹⁴?C activities of rice roots and rhizosphere soils in the L side were increased by intercropping with watermelon or AMF colonization. The ¹⁴?C was detected in roots and rhizosphere soils of rice and watermelon in the S side, but no differences were found among different treatments. ¹⁴?C activities in leaves were improved by AMF inoculation in the S side, regardless of rice or watermelon. Mycorrhizal colonization stimulated P absorption and translocation to rice in intercropping system. These findings suggest that AMF colonization could increase C transfer from rice to watermelon while intercropping with watermelon could promote AMF colonization and P uptake by rice.     

Der Umsatz von Pflanzenwurzeln im Laufe der Vegetationsperiode und dessen Beitrag zur „Bodenatmung”︁

The Turnover of Plant Roots during the Growth Period and its Influence on “Soll Respiration” Mustard and wheat plants were grown under ¹⁴CO₂, their roots being tightly separated from the shoot sphere. Root formation, root respiration, and root decomposition could thus be followed during the plant development by radiometric methods. The total quantity of organic root matter in soil at harvest time turned out to be 20–50% larger than the amount of root residues as determined by ordinary washing procedures. Depending on the plant and duration of the experiment, an additional amount of up to three times more than this remaining root carbon was already mineralized during the vegetation period. Only one fifth of this ¹⁴CO₂-production could be attributed to the respiration of living root tissue, all the remainder seemed to be due to the microbial decomposition of dead roots, root residues and root excretions. Root respiration and root decomposition together produced almost four fifths of the total evolving CO₂-quantity, whilst the contribution from soil organic matter breakdown did not exceed one fourth of it. According to these data, the total rhizo-deposition amounts to 3–4 times as much organic substance than what can be found as root residues at harvest time. This rich supply of readily decomposable organic matter leads to a most intensive turnover in the rhizosphere, which should be of considerable influence on the dynamic processes in 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.     

Determination of the fate of regularly applied 13C‐labeled‐artificial‐exudates C in two agricultural soils

Exudates are part of the total rhizodeposition released by plant roots to soil and are considered as a substantial input of soil organic matter. Exact quantitative data concerning the contribution of exudates to soil C pools are still missing. This study was conducted to reveal effects of ¹³C‐labeled exudate (artificial mixture) which was regularly applied to upper soil material from two agricultural soils. The contribution of exudate C to water‐extractable organic C (WEOC), microbial biomass C (MBC), and CO₂‐C evolution was investigated during a 74 d incubation. The WEOC, MBC, and CO₂‐C concentrations and the respective δ¹³C values were determined regularly. In both soils, significant incorporation of artificial‐exudate‐derived C was observed in the WEOC and MBC pool and in CO₂‐C. Up to approx. 50% of the exudate‐C amounts added were recovered in the order WEOC << MBC < CO₂‐C in both soils at the end of the incubation. Newly built microbial biomass consisted mainly of exudates, which substituted soil‐derived C. Correspondingly, the CO₂‐C evolved from exudate‐treated soils relative to the controls was dominated by exudate C, showing a preferential mineralization of this substrate. Our results suggest that the remaining 50% of the exudate C added became stabilized in non‐water‐extractable organic fractions. This assumption was supported by the determination of the total organic C in the soils on the second‐last sampling towards the end of the incubation. In the exudate‐treated soils, significantly more soil‐derived C compared to the controls was found in the WEOC on almost all samplings and in the MBC on the first sampling. This material might have derived from exchange processes between the added exudate and the soil matrix. This study showed that easily available substrates can be stabilized in soil at least in the short term.