Release of C and N from roots of peas and oats and their availability to soil microorganisms

Nutrient mobilisation in the rhizosphere is driven by soil microorganisms and controlled by the release of available C compounds from roots. It is not known how the quality of release influences this process in situ. Therefore, the present study was conducted to investigate the amount and turnover of rhizodeposition, in this study defined as root-derived C or N present in the soil after removal of roots and root fragments, released at different growth stages of peas (Pisum sativum L.) and oats (Avena sativa L.). Plants were grown in soil columns placed in a raised bed under outdoor conditions and simultaneously pulse labelled in situ with a ¹³C-glucose-¹⁵N-urea solution using a stem feeding method. After harvest, ¹³C and ¹⁵N was recovered in plant parts and soil pools, including the microbial biomass. Net rhizodeposition of C and N as a percentage of total plant C and N was higher in peas than in oats. Moreover, the C-to-N ratio of the rhizodeposits was lower in peas, and a higher proportion of the microbial biomass and inorganic N was derived from rhizodeposition. These results suggest a positive plant-soil feedback shaping nutrient mobilisation. This process is driven by the C and N supply of roots, which has a higher availability in peas than in oats.     

Fate of C- and N-labelled rhizodeposition of Lolium perenne as function of the distance to the root surface

A greenhouse rhizobox experiment was carried out to quantify the incorporation of ¹³C- and ¹⁵N-labelled rhizodeposits into different soil pools, especially into the rhizosphere microbial biomass, with increasing distances to the root surface of Lolium perenne. Five layers were analysed over 0-4.2 mm distance to an artificial root surface. C and N derived from rhizodeposition were 4.2% of total C and 2.8% of total N in soil at 0-1.0 mm distance and decreased rapidly with increasing distance. Microbial biomass C and N increased significantly towards the roots. At 0-1.0 mm distance microbial biomass C and N accounted for 66% and 29% of C and N derived from rhizodeposition, respectively. These percentages declined with increasing distance to the roots, but were still traceable up to 4.2 mm distance. Only small amounts of root released C and N were found in the 0.05 M K₂SO₄-extractable fraction. Extractable C and N derived from rhizodeposition varied around means of 4% of total C and N derived from rhizodeposition and increased only marginally with increasing distance to the roots. C derived from rhizodeposition in the non-extractable soil organic matter increased from 65 to 89% of total C derived from rhizodeposition at 0-3.4 mm distance. Conversely, microbial biomass C derived from rhizodeposition decreased from 33 to 4%. N derived from rhizodeposition in the non-extractable soil organic matter increased from 61 to 79% of total N derived from rhizodeposition at 0-2.6 mm distance, followed by a decline to roughly 55% in the two outer layers. Microbial biomass N decreased from 37 to 16% at 0-2.6 mm distance, followed by an increase to roughly 41% in the two outer layers. The C/N ratio of total C and N derived from rhizodeposition as well as that of extractable C and N derived from rhizodeposition increased with increasing distance to the roots to values above 30. In contrast, the C/N ratio of incorporated rhizodeposition C and N into the microbial biomass decreased to values less than 5 at 2.6-4.2 mm distance. The data indicate differential microbial response to C and N derived from rhizodeposition at a high spatial resolution from the root surface. The turnover of C and N derived from rhizodeposition in the rhizosphere as a function of the distance to the root surface is discussed.     

How to calculate nitrogen rhizodeposition: a case study in estimating N rhizodeposition in the pea (Pisum sativum L.) and grasspea (Lathyrus sativus L.) using a continuous N labelling split-root technique

The formulae used in studies with ¹⁵N labelling techniques for estimating the N rhizodeposition (Ndfr) of legumes differ according to the background atom% ¹⁵N values used to determine ¹⁵N excess in the soil and roots grown in soil. Therefore, a continuous ¹⁵N labelling split-root experiment with pea (Pisum sativum L.) and grasspea (Lathyrus sativus L.) was undertaken and the relevant calculations were made to determine a valid method for calculating Ndfr. It is shown that a non-nodulated reference plant or a legume grown on soil without ¹⁵N labelling are required components of experiments which aim to estimate legume-N rhizodeposition, if the ¹⁵N abundance of the total soil N at the start of the experiment and that of the total plant available soil N are different. The standard formula was developed further to calculate Ndfr in a valid way. The impact of using different background atom% ¹⁵N values on the results when estimating Ndfr are demonstrated according to the ¹⁵N abundance of the roots grown in the soil. At physiological maturity, the rhizodeposition of N from roots grown in the soil was 19.8 mg N plant⁻¹ for pea and 14.1 mg N plant⁻¹ for grasspea, which is, respectively, equivalent to 10.5 and 9.2% of their total root and shoot N.     

Nitrogen rhizodeposition in agricultural crops: Methods,estimates and future prospects

The objective of the present review was to present the current knowledge on nitrogen (N) rhizodeposition, including techniques for ¹⁵N labelling of agricultural plants, amounts of N rhizodeposition and its fate in soil. Rhizodeposition is the process of release of organic and inorganic compounds from living plant roots. It is often quantified in terms of carbon (C) and less often as N derived from rhizodeposition (NdfR). Rhizodeposition of N can be estimated by labelling plants with ¹⁵N and following its fate in soil. Most methods used for labelling plants with ¹⁵N can only be applied after appearance of the first leaf and only allow pulse or multiple pulse labelling. Only the split-root technique and the application of gaseous ¹⁵N allow continuous labelling. All methods available at present have their flaccidities mostly due to the fact that the application of N is not following its physiological pathway of assimilation or by using artificial conditions. In the studies reviewed, amounts of N rhizodeposits ranged from 4% to 71% of total assimilated plant N. In legumes the median was 16% and in cereals it was 14%. Rhizodeposits were 15–96% of the below-ground plant biomass (BGP). In legumes the median was 73% and in cereal it was 57%. The high variability of these results shows the need for more investigations on N rhizodeposition looking especially on the factors influencing the amounts released in different plant species under field conditions.     

Does labelling frequency affect N rhizodeposition assessment using the cotton-wick method?

The aim of the present study was to test and improve the reliability of the ¹⁵N cotton-wick method for measuring soil N derived from plant rhizodeposition, a critical value for assessing belowground nitrogen input in field-grown legumes. The effects of the concentration of the ¹⁵N labelling solution and the feeding frequency on assessment of nitrogen rhizodeposition were studied in two greenhouse experiments using the field pea (Pisum sativum L.). Neither the method nor the feeding frequency altered plant biomass and N partitioning, and the method appeared well adapted for assessing the belowground contribution of field-grown legumes to the soil N pool. However, nitrogen rhizodeposition assessment was strongly influenced by the feeding frequency and the concentration of labelling solution. At pod-filling and maturity, despite similar root ¹⁵N enrichment, the fraction of plants' belowground nitrogen allocated to rhizodeposition in both Frisson pea and the non-nodulating isoline P2 was 20 to more than 50% higher when plants were labelled continuously than when they were labelled using fortnightly pulses. Our results suggest that when ¹⁵N root enrichment was high, nitrogen rhizodeposition was overestimated only for plants that were ¹⁵N-fed by fortnightly pulses, and not in plants ¹⁵N-fed continuously. This phenomenon was especially observed for plants that rely on symbiotic N₂ fixation for N acquisition, and it may be linked to the concentration of the labelling solution. In conclusion, the assessment of nitrogen rhizodeposition was more reliable when plants were labelled continuously with a dilute solution of ¹⁵N urea.     

Estimating N rhizodeposition of grain legumes using a N in situ stem labelling method

Grain legumes in crop rotations cause significant increases in yield for succeeding non-legumes, which cannot be explained simply by the small effect that legumes have on the soil nitrogen balance, as found in the analysis of N in crop residues. Besides known positive non-N-effects, other effects, mainly rhizodeposition and its contribution to the N balance and nitrogen dynamics after harvesting the grain, are poorly understood. In this study, N rhizodeposition, defined as root-derived N in the soil after removal of visible roots, was measured in faba bean (Vicia faba L.), pea (Pisum sativum L.) and white lupin (Lupinus albus L.). In a pot experiment the legumes were pulse labelled in situ with ¹⁵N urea using a cotton wick method. About 84% of the applied ¹⁵N was recovered for the three legume species at maturity. The ¹⁵N was comparatively uniformly distributed among plant parts. The N rhizodeposition constituted 13% of total plant N for faba bean and pea and 16% for white lupin at maturity, about 80% of below ground plant N, respectively. Some 7% (lupin)-31% (pea) of the total N rhizodeposits were recovered as micro-roots by wet sieving (200 μm) the soil after all visible roots had been removed. Only 14-18% of the rhizodeposition N was found in the microbial biomass and a very small amount of 3-7% was found in the mineral N fraction. In pea, 48% and in lupin 72% of N rhizodeposits could not be recovered in the mentioned pools and a major part of the unrecovered N was probably immobilised in microbial residues. The results of this study clearly indicate that N rhizodeposition from grain legumes represent a significant pool for N balance and N dynamics in crop rotations.     

Rhizodeposition of C and N in peas and oats after C-N double labelling under field conditions

Compounds released by plant roots during growth can make up a high proportion of below-ground plant (BGP) carbon and nitrogen, and therefore influence soil organic matter turnover and plant nutrient availability by stimulating the soil microorganisms. The present study was conducted to examine the amount and fate of C (CdfR) and N rhizodeposits (NdfR), in this study defined as root-derived C or N present in the soil after removal of roots and root fragments, released during reproductive growth. BGP biomass of peas (Pisum sativum L.) and oats (Avena sativa L.) was successfully labelled in situ with a ¹³C-glucose-¹⁵N-urea mixture under field conditions using a stem feeding method. Pea plants were labelled at the beginning of flowering and harvested 36 and 52 days after labelling at pod filling (P_P) and maturity (P_M), respectively. Oat plants were labelled at grain filling and harvested 42 days after labelling at maturity (O_M). CdfR was 24.2% (P_P), 29.6% (P_M) and 30.8% (O_M) of total recovered plant C. NdfR was 32.1% (P_P), 36.4% (P_M) and 30.0% (O_M) of total plant N. Due to higher N assimilation, amounts of NdfR were four times higher in peas in comparison with oats. The results for NdfR in peas were higher than results from other studies. The C-to-N ratio of rhizodeposits was lower under peas (17.3) than under oats (41.9) at maturity. At maturity, microbial CdfR at 0-30 cm soil depth was 37% of the microbial biomass C in peas and 59% in oats. Microbial NdfR was 15% of microbial N in peas and 5% in oats. Furthermore, inorganic NdfR was 34% in peas and 9% in oats at 0-30 cm at maturity. These results show that rhizodeposits of peas provide a more easily available substrate to soil microorganisms, which are incorporated to a greater extent and turned over faster in comparison with oats. Beside the higher amounts of N released from pea roots, this process contributes to the higher N-availability for subsequent crops.     

Turnover of grain legume N rhizodeposits and effect of rhizodeposition on the turnover of crop residues

The turnover of N derived from rhizodeposition of faba bean (Vicia faba L.), pea (Pisum sativum L.) and white lupin (Lupinus albus L.) and the effects of the rhizodeposition on the subsequent C and N turnover of its crop residues were investigated in an incubation experiment (168 days, 15 °C). A sandy loam soil for the experiment was either stored at 6 °C or planted with the respective grain legume in pots. Legumes were in situ ¹⁵N stem labelled during growth and visible roots were removed at maturity. The remaining plant-derived N in soil was defined as N rhizodeposition. In the experiment the turnover of C and N was compared in soils with and without previous growth of three legumes and with and without incorporation of crop residues. After 168 days, 21% (lupin), 26% (faba bean) and 27% (pea) of rhizodeposition N was mineralised in the treatments without crop residues. A smaller amount of 15–17% was present as microbial biomass and between 30 and 55% of mineralised rhizodeposition N was present as microbial residue pool, which consists of microbial exoenzymes, mucous substances and dead microbial biomass. The effect of rhizodeposition on the C and N turnover of crop residues was inconsistent. Rhizodeposition increased the crop residue C mineralisation only in the lupin treatment; a similar pattern was found for microbial C, whereas the microbial N was increased by rhizodeposition in all treatments. The recovery of residual ¹⁵N in the microbial and mineral N pool was similar between the treatments containing only labelled crop residues and labelled crop residues + labelled rhizodeposits. This indicates a similar decomposability of both rhizodeposition N and crop residue N and may be attributable to an immobilisation of both N sources (rhizodeposits and crop residues) as microbial residues and a subsequent remineralisation mainly from this pool.Abbreviations C or N_dec C or N decomposed from residues - C or N_mic microbial C or N - C or N_micres microbial residue C or N - C or N_min mineralised C or N - C or N_input added C or N as crop residues and/or rhizodeposits - dfr derived from residues - dfR derived from rhizodeposition - Ndfr N derived from residues - NdfR N derived from rhizodeposition - N_loss losses of N derived from residues - SOM soil organic matter - WHC water holding capacity     

Rhizodeposition: Its contribution to microbial growth and carbon and nitrogen turnover within the rhizosphere

A greenhouse rhizobox experiment was carried out to investigate the fate and turnover of ¹³C‐ and ¹⁵N‐labeled rhizodeposits within a rhizosphere gradient from 0–8 mm distance to the roots of wheat. Rhizosphere soil layers from 0–1, 1–2, 2–3, 3–4, 4–6, and 6–8 mm distance to separated roots were investigated in an incubation experiment (42 d, 15°C) for changes in total C and N and that derived from rhizodeposition in total soil, in soil microbial biomass, and in the 0.05 M K₂SO₄–extractable soil fraction. CO₂‐C respiration in total and that derived from rhizodeposition were measured from the incubated rhizosphere soil samples. Rhizodeposition C was detected in rhizosphere soil up to 4–6 mm distance from the separated roots. Rhizodeposition N was only detected in the rhizosphere soils up to 3–4 mm distance from the roots. Microbial biomass C and N was increased with increasing proximity to the separated roots. Beside ¹³C and ¹⁵N derived from rhizodeposits, unlabeled soil C and N (native SOM) were incorporated into the growing microbial biomass towards the roots, indicating a distinct acceleration of soil organic matter (SOM) decomposition and N immobilization into the growing microbial biomass, even under the competition of plant growth. During the soil incubation, microbial biomass C and N decreased in all samples. Any decrease in microbial biomass C and N in the incubated rhizosphere soil layers is attributed mainly to a decrease of unlabeled (native) C and N, whereas the main portion of previously incorporated rhizodeposition C and N during the plant growth period remained immobilized in the microbial biomass during the incubation. Mineralization of native SOM C and N was enhanced within the entire investigated rhizosphere gradient. The results indicate complex interactions between substrate input derived from rhizodeposition, microbial growth, and accelerated C and N turnover, including the decomposition of native SOM (i.e., rhizosphere priming effects) at a high spatial resolution from the roots.     

Rhizodeposition of nitrogen by red clover,white clover and ryegrass leys

Correct assessment of the rhizodeposition of N in grassland is essential for the evaluation of biological N₂-fixation of legumes, for the total N balance of agro-ecosystems, and for the pre-cropping value of grasslands. Using a leaf-feeding technique by which plants were ¹⁵N labelled while growing in mezotrons in the field, the rhizodeposition of N by unfertilised red clover, white clover and perennial ryegrass growing in pure stands was shown to amount to 64, 71 and 9 g N m⁻², respectively, over two complete growing seasons. The corresponding values for red clover and white clover growing in mixtures with ryegrass were 89 and 32 g N m⁻², respectively. The rhizodeposited N compounds, including fine roots, constituted more than 80% of the total plant-derived N in the soil, and in all cases exceeded the amount of N present in stubble. In the mixtures of red clover–ryegrass and white clover–ryegrass and the pure stands of red clover, white clover and ryegrass, respectively, the rhizodeposition constituted a 1.05, 1.52, 1.26, 2.21 and 2.77 fold increase over the total N in the shoots harvested during the two production years. In pure stands and mixtures of clover, 84 and 92%, respectively, of this N derived from biological N₂ fixation. It is concluded that rhizodeposition provides a very substantial input of N to the legume-based grassland systems with great consequences for ecosystem N balance and turnover. Furthermore, the amount of atmospheric-derived N in the rhizodeposits may exceed that in the harvested shoots.     

A model to predict the accuracy of measurements of legume N rhizodeposition using a split-root technique

Legume N rhizodeposition is an important process for understanding the N turnover in legume-based cropping systems. Different ¹⁵N labeling techniques have been developed to estimate the rhizodeposition of legume-derived N into the soil. However, it is not known how the ¹⁵N-based experiments have to be designed to achieve a defined degree of accuracy in measuring the amount of N derived from rhizodeposition (Ndfr). As a consequence therefore, a model for the split-root technique was developed on the basis of experimental data to (i) test the effects of various experimental conditions on soil ¹⁵N enrichment, (ii) evaluate the accuracy of the measurements, and to (iii) deduce instructions for designing efficient experiments using ¹⁵N techniques for quantifying legume N rhizodeposition. It turned out that the coefficient of disproportional ¹⁵N distribution between non-¹⁵N-fed roots grown in soil (Root_Soil) and total plant biomass (D coefficient) is an indispensable component in the development of such a model for ¹⁵N-based split-root experiments. The model showed the coefficients of variation in measuring the Ndfr with regard to the analytical accuracy in determining not only the isotope composition of both the Root_Soil and the soil, but also the N content of the soil itself. Suggestions for designing specific experimental conditions to achieve a high accuracy in quantifying Ndfr were deduced from the model for the split-root technique, particularly in the choice of the amount of soil N at the start of the experiment and the ¹⁵N enrichment of the fertilizer being used. The coefficients of variation in measuring Ndfr are presented regarding the ¹⁵N abundance of Root_Soil and the quotient of the amount of Ndfr and soil N at the start of the experiment.     

土施和枝干涂抹 15N-尿素对氮素在甜樱桃树不同部位分配及利用的影响

大田条件下利用15N示踪技术,研究了土施和枝干涂抹15N-尿素对氮素在甜樱桃树不同部位分配及利用的影响。结果表明: 果实采收期测定两种不同施肥方式各器官的Ndff%存在差异,土壤施肥处理细根的Ndff%最高为2.36%; 枝干涂抹处理短梢的Ndff%最高为4.26%。土壤施肥处理粗根的15N分配率最高为22.23%,其次为短梢叶和果实; 枝干涂抹处理中心干皮部的15N分配率最高为26.14%,其次为中心干木质部和果实。两种不同施肥方式下植株营养器官和贮藏器官的15N分配率差异极显著。果实采收期测定枝干涂抹处理和土壤施肥处理的15N利用率分别为20.64%和14.74%。     

Collembola switch diet in presence of plant roots thereby functioning as herbivores

Collembola are abundant and ubiquitous soil decomposers, being particularly active in the rhizosphere of plants where they are assumed to be attracted by high microbial activity and biomass. While feeding on root associated microorganisms or organic matter they may also ingest plant roots, e.g. particularly root hairs and fine roots. Employing stable isotope analysis we investigated Collembola (Protaphorura fimata Gisin) feeding preferences and types of ingested resources. We offered Collembola two resources with distinct isotope signatures: a C4 plant (Zea mays L.) planted in soil mixed with ¹⁵N labelled litter of Lolium perenne L. (C3 plant). We hypothesised that Collembola obtain their nutrients (C and N) from different resources, with their carbon being mainly derived from resources that are closely associated to the plant root, e.g. root exudates, causing enrichment in ¹³C in Collembola tissue, while the incorporated nitrogen originating from litter resources. In contrast to our hypothesis, stable isotope analysis suggests that in absence of plant roots Collembola derived both the incorporated C and N predominantly from litter whereas in presence of plant roots they switched diet and obtained both C and N almost exclusively from plant roots.The results indicate that Collembola in the rhizosphere of plants, being assumed to be mainly decomposers, in fact predominately live on plant resources, presumably fine roots or root hairs, i.e. are herbivorous rather than detritivorous or fungivorous. These findings have major implications on the view how plants respond to decomposers in the rhizosphere.     

Rice rhizodeposition and its utilization by microbial groups depends on N fertilization

Rhizodeposits have received considerable attention, as they play an important role in the regulation of soil carbon (C) sequestration and global C cycling and represent an important C and energy source for soil microorganisms. However, the utilization of rhizodeposits by microbial groups, their role in the turnover of soil organic matter (SOM) pools in rice paddies, and the effects of nitrogen (N) fertilization on rhizodeposition are nearly unknown. Rice (Oryza sativa L.) plants were grown in soil at five N fertilization rates (0, 10, 20, 40, or 60 mg N kg^?1 soil) and continuously labeled in a ¹³CO₂ atmosphere for 18 days during tillering. The utilization of root-derived C by microbial groups was assessed by ¹³C incorporation into phospholipid fatty acids. Rice shoot and root biomass strongly increased with N fertilization. Rhizodeposition increased with N fertilization, whereas the total ¹³C incorporation into microorganisms, as indicated by the percentage of ¹³C recovered in microbial biomass, decreased. The contribution of root-derived ¹³C to SOM formation increased with root biomass. The ratio of ¹³C in soil pools (SOM and microbial biomass) to ¹³C in roots decreased with N fertilization showing less incorporation and faster turnover with N. The ¹³C incorporation into fungi (18:2ω6,9c and 18:1ω9c), arbuscular mycorrhizal fungi (16:1ω5c), and actinomycetes (10Me 16:0 and 10Me 18:0) increased with N fertilization, whereas the ¹³C incorporation into gram-positive (i14:0, i15:0, a15:0, i16:0, i17:0, and a17:0) and gram-negative (16:1ω7c, 18:1ω7c, cy17:0, and cy19:0) bacteria decreased with N fertilization. Thus, the uptake and microbial processing of root-derived C was affected by N availability in soil. Compared with the unfertilized soil, the contribution of rhizodeposits to SOM and microorganisms increased at low to intermediate N fertilization rates but decreased at the maximum N input. We conclude that belowground C allocation and rhizodeposition by rice, microbial utilization of rhizodeposited C, and its stabilization within SOM pools are strongly affected by N availability: N fertilization adequate to the plant demand increases C incorporation in all these polls, but excessive N fertilization has negative effects not only on environmental pollution but also on C sequestration in soil.     

Relationships between C and N availability, substrate age, and natural abundance C and N signatures of soil microbial biomass in a semiarid climate

Soil microbial organisms are central to carbon (C) and nitrogen (N) transformations in soils, yet not much is known about the stable isotope composition of these essential regulators of element cycles. We investigated the relationship between C and N availability and stable C and N isotope composition of soil microbial biomass across a three million year old semiarid substrate age gradient in northern Arizona. The δ¹⁵N of soil microbial biomass was on average 7.2‰ higher than that of soil total N for all substrate ages and 1.6‰ higher than that of extractable N, but not significantly different for the youngest and oldest sites. Microbial ¹⁵N enrichment relative to soil extractable and total N was low at the youngest site, increased to a maximum after 55,000 years, and then decreased slightly with age. The degree of ¹⁵N enrichment of microbial biomass correlated negatively with the C:N mass ratio of the soil extractable pool. The δ¹³C signature of soil microbial biomass was 1.4‰ and 4.6‰ enriched relative to that of soil total and extractable pools respectively and showed significant differences between sites. However, microbial ¹³C enrichment was unrelated to measures of C and N availability. Our results confirm that ¹⁵N, but not ¹³C enrichment of soil microbial biomass reflects changes in C and N availability and N processing during long-term ecosystem development.     

Comparing N-labelling techniques for enriching above- and below-ground components of the plant-soil system

Comparisons were made of three different ¹⁵N-feeding techniques, leaf, petiole and stem feeding, to identify the most efficient technique for labelling above-and below-ground plant biomass under controlled environment conditions. ¹⁵N-urea (0.5%, 10 atom % excess ¹⁵N) was applied to chickpea (Cicer aritenium var. ICCV 5003) plants twice during early growth. Leaf feeding was found to be the most efficient in terms of ¹⁵N-solution uptake (5.9 ml 48 h⁻¹) and ¹⁵N-enrichment at harvest, with 0.95, 0.41, 0.79, 0.67 and 0.22 atom % excess ¹⁵N in the leaves, stems, grain, grain straw and clean root fractions, respectively. Solution uptake was low in the second stem feeding event due to blockage of the drilled hole, resulting in low ¹⁵N-enrichment of leaves (0.29 atom % excess ¹⁵N). Although petiole feeding resulted in more even relative enrichments among plant parts our results highlight the usefulness of leaf ¹⁵N-feeding to estimate below-ground plant N and to trace the long-term fate of plant-derived N within the soil.     

Nitrogen rhizodeposition of young wheat plants under elevated CO<Subscript>2</Subscript> and drought stress

The objective of this study was to determine the effect of drought stress and elevated CO₂ concentrations around the shoots on N rhizodeposition of young wheat plants. In a pot experiment, the plant N pool was labeled through ¹⁵NH₃ application to shoots at nontoxic NH₃ concentrations, and the impact of low water supply (40% field capacity), elevated CO₂ (720 μmol mol⁻¹ CO₂), and the combination of both factors on the ¹⁵N distribution was studied. Total ¹⁵N rhizodeposition ranged from 5 to 11% of the total ¹⁵N recovered in the plant/soil system. Elevated CO₂ concentration as well as drought stress increased the belowground transport of N and increased the relative portion of N rhizodeposition on total ¹⁵N in the plant/soil system. However, while the increased N rhizodeposition with elevated CO₂ was the result of increased total belowground N transport, drought stress additionally increased the portion of ¹⁵N found in rhizodeposition vs roots. Elevated CO₂ intensified the effect of drought stress. The percentage of water soluble ¹⁵N in the ¹⁵N rhizodeposition was very low under all treatments, and it was significantly decreased by the drought-stressed treatments.     

A comparison of different 15N application techniques to study the N net rhizodeposition in the plant‐soil system

The suitability of three ¹⁵N application methods (¹⁵NH₃ fumigation, split‐root technique, ¹⁵N pre‐cultivation) for the estimation of N net rhizodeposition (NRD) of wheat plants into soil has been tested and compared under similar conditions and at the same developmental stage. The results were as follows: 1. The use of the ¹⁵N tracer technique allows the detection of the net N release by roots under soil conditions. NRD was considerable and can be estimated to be at least 15 kg N ha⁻¹ a⁻¹. 2. All three methods applied are practicable under non‐sterile experimental conditions. The distribution of applied ¹⁵N in the system and NRD can be balanced totally only by using the ¹⁵NH₃ fumigation and the ¹⁵N pre‐cultivation methods. The split‐root technique leads to an overestimation of NRD. 3. The split‐root technique allows a qualitative separation of the NRD under nearly undisturbed conditions. With the ¹⁵N precultivation, a higher ¹⁵N‐labelling can be achieved for long‐term balance studies. 4. Despite the required high ¹⁵N abundance, the ¹⁵NH₃ fumigation method works best to evaluate the influence of microbes on NRD and to quantify the gaseous ¹⁵N release.     

Interspecies competition and N transfer in a tropical grass-legume mixture

Competitiveness of Brachiaria decumbens cv. Basilisk and Stylosanthes guianensis cv. Minerão was investigated either without root restriction or by separating their root systems with a fine mesh or a solid barrier in the presence or absence of mycorrhiza (Glomus clarum). Nitrogen transfer between the legume and the grass was assessed with the ¹⁵N isotope dilution technique using a relatively stable ¹⁵N-enriched soil derived from a long-term labelling experiment. During establishment, legume development was severely restricted by competition from the grass in pots without a root barrier. However, as the system became N limited, the legume became dominant due to its access to atmospheric N₂ which contributed over 80% of the legume N requirements. S. guianensis was highly mycotrophic and inoculation with mycorrhiza favoured rapid establishment even in the treatments with no root barrier. Only in the presence of root barriers, either a mesh or a complete compartment separation, was the proportion of N derived from N₂ fixation positively affected by the presence of the fungus. No significant direct belowground N transfer from legume to grass was observed during the lifetime of the legume suggesting that the legume maintains a highly efficient recycling under N-limited conditions. However, after cutting the shoot at ground level, the grass assimilated significant amounts of N derived from decaying legume roots. We conclude that the main pathway of belowground N transfer from S. guianensis to associated B. decumbens occurred via decomposing roots rather than via root exudates or direct mycorrhizal hyphae transfer.     

Alleviation of P limitation makes tree roots competitive for N against microbes in a N-saturated conifer forest: A test through P fertilization and 15N labelling

Chronic N deposition to forests may induce N saturation and stand decline, leading to reduced ecosystem N retention capacity, triggered by a shift from N limitation of trees to limitation by another nutrient. We conducted a ¹⁵N soil labelling experiment in non-fertilized and P-fertilized plots at two elevations in an N-saturated Mediterranean-fir (Abies pinsapo) forest in southern Spain which shows P limitation symptoms. Root-exclusion was applied to identify the relative contributions of roots (plus mycorrhizal fungi) uptake, and heterotrophic immobilization by free-living microbes, to N retention. Overall ¹⁵N recovery from the litter, 0–15-cm soil and root-uptake components was c.a. 35% higher in P-fertilized than in non-fertilized plots at both elevations. In non-fertilized plots, soil was the biggest sink for added ¹⁵N. Phosphorus fertilization increased the competitive ability of tree roots for soil N resulting in equal importance of the autotrophic (roots plus associated mycorhizal fungi) and heterotrophic (free-living microbes) components with respect to total ¹⁵N recovery in P-fertilized plots. Phosphorus addition increased litter and soil N immobilization only if roots had been excluded. By combining in situ fertilization, root-exclusion and isotope labelling we have demonstrated that reduced N retention capacity and dominance of soil microbial over plant immobilization in a N-saturated forest results from a shift from N to P limitation of trees, while alleviation of P limitation makes tree roots and associated mycorrhizal fungi competitive for N against free soil microorganisms.