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.     

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.     

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.     

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     

Quantitative evidence of overestimated rhizodeposition using 15N leaf-labelling

Nitrogen (N) rhizodeposition is defined as the release of N from living plant roots into the soil and root turnover. The proportion of N in the soil derived from rhizodeposition (NdfR) is usually determined using ¹⁵N labelling of the plant. This isotope approach assumes that i) the enrichment of rhizodeposits is equal to the root enrichment and that ii) the root remains homogeneously enriched over space and iii) over time. The aim of this study was to quantify the bias resulting from a possible violation of the mentioned assumptions and to study the causative factors of bias.We conducted two experiments with single-pulse ¹⁵N-urea leaf-labelled red clover (Trifolium pratense L.). In the rhizodeposition experiment, we simultaneously observed the changes in substrate N concentration to obtain the mass-based rhizodeposits and determined the isotope-predicted rhizodeposits using the isotope approach. By comparing isotope-predicted to mass-based rhizodeposits we quantified the bias of the isotope approach for a period of 6 weeks. In the root distribution experiment, we observed the root ¹⁵N enrichment over space and time (4 weeks) by sampling roots grown within certain periods relative to the labelling. In both experiments we monitored the ¹⁵N distribution between shoots and roots.We observed violations of the three assumptions of the isotope approach. The average root enrichment increased over time in the root distribution experiment, but remained constant in the rhizodeposition experiment. Significant long-term translocation of ¹⁵N from shoot to root during the whole experiment (over)-compensated for growth dilution. Spatial root enrichment varied within a factor of 3, peaking in roots grown 2–8 days after labelling (d.a.l.). We observed a significant leakage of 0.5 ± 0.2% of the applied ¹⁵N within the first day after labelling corresponding to an overestimation of first-day rhizodeposits by 1100 ± 800% which translates to a calculated enrichment of 9 ± 6 atom% ¹⁵N excess for first-day rhizodeposits compared to 0.77 ± 0.09 atom% ¹⁵N excess of the root.The leaked ¹⁵N together with ordinary rhizodeposits (rhizodeposits released after the first day of labelling) led to an overestimation of N rhizodeposition by 70 ± 30% at the end of the six weeks lasting experiment using the isotope approach. The observed ¹⁵N distribution in roots and long-term ¹⁵N translocation from shoots to roots did not correspond to the expected distribution following classical pulse labelling. Thus, leaf-labelling with ¹⁵N-urea should not be considered a pure pulse-labelling method.     

Stem labeling results in different patterns of 14C rhizorespiration and 15N distribution in plants compared to natural assimilation pathways

To investigate C and N rhizodeposition, plants can be ¹³C‐¹⁵N double‐labeled with glucose and urea using a stem‐feeding method (wick method). However, it is unclear how the ¹³C applied as glucose is released into the soil as rhizorespiration in comparison with the ¹³C applied as CO₂ using a natural uptake pathway. In the present study, we therefore compared the short‐term fate of ¹⁴C and ¹⁵N in white lupine and pea plants applied either by the wick method or the natural pathways of C and N assimilation. Plants were pulse‐labeled in ¹⁴CO₂‐enriched atmosphere and ¹⁵N urea was applied to the roots (atmosphere–soil) following the natural assimilation pathways, or plants were simultaneously labeled with ¹⁴C and ¹⁵N by applying a ¹⁴C glucose–¹⁵N urea solution into the stem using the wick method. Plant development, soil microbial biomass, total rhizorespiration, and distribution of N in plants were not affected by the labeling method used but by plant species. However, the ¹⁵N : N ratio in plant parts was significantly (p < 0.05) affected by the labeling method, indicating more homogeneous ¹⁵N enrichment of plants labeled via root uptake. After ¹⁴CO₂ atmosphere labeling of plants, the cumulated ¹⁴CO₂ release from roots and soil showed the common saturation dynamics. In contrast, after ¹⁴C‐glucose labeling by the wick method, the cumulated ¹⁴CO₂ release increased linearly. These results show that ¹⁴C applied as glucose using the wick method is not rapidly transferred to the roots as compared to a short‐term ¹⁴CO₂ pulse. This is partly due to a slower ¹⁴C uptake and partly due to slow distribution within the plant. Consequently, ¹⁴C‐glucose application by the wick method is no pulse‐labeling approach. However, the advantages of the wick method for ¹³C‐¹⁵N double labeling for estimating rhizodeposition especially under field conditions requires further methodological research.     

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 15N tracing technique-based analysis of the fate of fertilizer N: a 4-year case study in eastern China

The fate of fertilizer N applied with different irrigation amounts in tobacco fields was quantitatively studied by applying ¹⁵N double-labelled NH₄NO₃ in lysimeters. The ¹⁵N (fertilizer N originating from the fertilizer applied in 2011) in tobacco plants, ¹⁵N in soils and ¹⁵N loss were observed continuously from 2011 to 2014. The results showed that 21.6% of ¹⁵N was utilized by tobacco plants, 72.1% remained in the 0–60 cm soil layer and 6.3% was lost from the soil–plant system after the first season’s harvest (2011) of flue-cured tobacco. During the four seasons from 2011 to 2014, cumulative utilization of ¹⁵N by tobacco plants was 34.3%, while 54.2% remained in the 0–60 cm soil layer, and 11.5% was lost via mechanisms such as leaching and volatilization. The fate of ¹⁵N in terms of accumulation in plants and soils or losses from the soil–plant system from 2012 to 2014 was greatly affected by the fertilizer and irrigation management strategies in 2011. The results of this investigation suggest that the major amount of fertilizer N applied during the first season remains available in the soil for utilization by tobacco plants after 4 years.     

Effects of Multiple Reference Plants,Season, and Irrigation on Biological Nitrogen Fixation by Pasture Legumes using the Isotope Dilution Method

Abstract

The popular and widely used ¹⁵nitrogen (N)–isotope dilution method for estimating biological N fixation (BNF) of pasture and tree legumes relies largely on the ability to overcome the principal source of error due to the problem of selecting appropriate reference plants. A field experiment was conducted to evaluate the suitability of 12 non‐N₂‐fixing plants (i.e., nonlegumes) as reference plants for estimating the BNF of three pasture legumes (white clover, Trifolium repens L.; lucerne, Medicago sativa; and red clover, Trifolium pratense L.) in standard ryegrass–white clover (RWC) and multispecies pastures (MSP) under dry‐land and irrigation systems, over four seasons in Canterbury, New Zealand. The ¹⁵N‐isotope dilution method involving field ¹⁵N‐microplots was used to estimate BNF. Non‐N₂‐fixing plants were used either singly or in combination as reference plants to estimate the BNF of the three legumes. Results obtained showed that, on the whole, ¹⁵N‐enrichment values of legumes and nonlegumes varied significantly according to plant species, season, and irrigation. Grasses and herb species showed higher ¹⁵N‐enrichment than those of legumes. Highest ¹⁵N‐enrichment values of all plants occurred during late summer under dry‐land and irrigation conditions. Based on single or combined non‐N₂‐fixing plants as reference plants, the proportion of N derived from the atmosphere (% Ndfa) values were high (50 to 90%) and differed between most reference plants in the MSP pastures, especially chicory (Cichorium intybus), probably because it is different in phenology, rooting depth, and N‐uptake patterns compared to those of legumes. The percent Ndfa values of all plants studied also varied according to plant species, season, and irrigation in the MSP pastures. Estimated daily amounts of BNF varied according to pasture type, time of plant harvest, and irrigation, similar to those shown by percent Ndfa results as expected. Irrigation increased daily BNF more than 10‐fold, probably due to increased dry‐matter yield of pasture under irrigation compared to dry‐land conditions. Seasonal and irrigation effects were more important in affecting estimates of legume BNF than those due to the appropriate matching of N₂‐fixing and non‐N₂‐fixing reference plants.     

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.     

Assessment of the relative uptake of added and indigenous soil nitrogen by nodulated legumes and reference plants in the 15N dilution measurement of N2 fixation: Derivation of method

Current methods for measuring N₂ fixation by nodulated legumes involve the addition of small amounts of ¹⁵N-labelled plant-available N compounds to soil so that plant N derived from the soil may be identified. All such methods assume that the proportions of added N and indigenous soil N assimilated by N₂-fixing and non-fixing plants grown in the same soil are the same, irrespective of the amount of soil N assimilated. The development of a method for assessing these proportions is described.Nodulated legumes and reference plants are grown in soils receiving none or one rate of addition of labelled N compound containing several (two or more) concentrations of ¹⁵N. The proportions of added and indigenous N assimilated, are determined from the intercepts and slopes of regression lines relating isotopic composition of plant N to that of added N, together with some other readilly-obtainable plant N measurements.     

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.     

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.     

Measuring the rhizodeposition of carbon by rice: an approach based on carbon flux observations

ABSTRACT

Rhizodeposition is an important component of carbon cycling in terrestrial ecosystems. However, there remains tremendous uncertainty in its quantification due to the methodological limitations. In the present study, we propose a method to evaluate the rhizodeposition by plants by observing carbon flux. We investigated the ecosystem CO₂ flux variability and calculated the rhizodeposition of carbon by the rice rhizosphere, by using the carbon flux, meteorological data, and biomass observation from 2003 to 2011 at the Taoyuan Agro-ecological Experimental Station, a representative subtropical paddy ecosystem. Our data indicated that the process of rhizodeposition is the major reason for the discrepancy between the biomass and net primary productivity of the paddy ecosystem under intensive human interference. Both the amount and ratio of rhizodeposition of carbon in this paddy ecosystem were assessed; this provides important theoretical and methodological support for further investigating rhizodeposition by rice under field conditions. The rhizodeposition amount in the growing season of early rice, late rice, and for the entire planting period was 0.52–2.56, 0.74–3.75, and 1.61–5.24 t ha^?1, respectively, with the corresponding mean (±SD) rhizodeposition ratios of 23.16 ± 8.87%, 28.16 ± 12.94%, and 27.00 ± 9.3%. This method enabled us to calculate rhizodeposition under in situ conditions, and the results showed that the growing season of late rice was the primary period for rhizodeposition in rice ecosystem.     

15N studies on the transfer of legume-fixed nitrogen to associated cereals in intercropping systems

Summary An attempt has been made to estimate quantitatively the amount of N fixed by legume and transferred to the cereal in association in intercropping systems of wheat (Triticum aestivum L.) — gram (Cicer arietinum L.) and maize (Zea mays L.) —cowpea (Vigna unguiculate L.) by labelling soil and fertilizer nitrogen with ¹⁵N. The intercropped legumes have been found to fix significantly higher amounts of N as compared with legumes in sole cropping if the intercropped cereal-legume received the same dose of fertilizer N as the sole cereal crop. But when half of the dose of the fertilizer N applied to sole cereal crop was received by intercropped plants, the amount of N fixed by legumes in association with cereals was significantly less than that fixed by sole legumes. Under field conditions 28% of the total N uptake by maize (21.2 kg N ha^–1) was of atmospheric origin and was obtained by transfer of fixed N by cowpea grown in association with maize. Under greenhouse conditions gram and summer and monsoon season cowpea have been found to contribute 14%–20%, 16% and 32% of the total N uptake by associated wheat and summer and monsoon maize crops, respectively. Inoculation of cowpea seeds with Rhizobium increased both the amount of N fixed by cowpea and transferred to maize in intercropping system.     

Incorporation of 13C-labelled rice rhizodeposition carbon into soil microbial communities under different water status

The overall processes by which carbon is fixed by plants in photosynthesis then released into the soil by rhizodeposition and subsequently utilized by soil micro-organisms, links the atmospheric and soil carbon pools. The objective of this study was to determine the plant derived ¹³C incorporated into the phospholipid fatty acid (PLFA) pattern in paddy soil, to test whether utilization of rice rhizodeposition carbon by soil micro-organisms is affected by soil water status. This is essential to understand the importance of flooded conditions in regulating soil microbial community structure and activity in wetland rice systems. Rice plants were grown in soil derived from a paddy system under controlled irrigation (CI), or with continuous waterlogging (CW). Most of the ¹³C-labelled rice rhizodeposition carbon was distributed into the PLFAs 16:0, 18:1ω7 and 18:1ω9 in both the CW and CI treatments. The bacterial PLFAs i15:0 and a15:0, both indicative of gram positive bacteria, were relatively more abundant in the treatments without rice plants. When rice plants were present rates of ¹³C-incorporation into i15:0 and a15:0 was slow; the microbes containing these PLFAs may derive most of their carbon from more recalcitrant C (soil organic matter). PLFAs, 18:1ω7 and 16:1ω7c, indicative of gram negative bacteria showed a greater amount incorporation of labelled plant derived carbon in the CW treatment. In contrast, 18:2ω6,9 indicative of fungi and 18:1ω9 indicative of aerobes but also potentially fungi and plant roots had greater incorporation in the CI treatment. The greater root mass concomitant with lower incorporation of ¹³C into the total PLFA pool in the CW treatment suggests that the microbial communities in wetland rice soil are limited by factors other than substrate availability in flooded conditions. In this study differing soil microbial communities were established through manipulating the water status of paddy soils. Steady state ¹³C labelling enabled us to determine that the microbial community utilizing plant derived carbon was also affected by water status.     

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.     

Dry-rewetting cycles regulate wheat carbon rhizodeposition,stabilization and nitrogen cycling

Drying and rewetting of soil can have large effects on carbon (C) and nitrogen (N) dynamics. Drying-rewetting effects have mostly been studied in the absence of plants, although it is well known that plant–microbe interactions can substantially alter soil C and N dynamics. We investigated for the first time how drying and rewetting affected rhizodeposition, its utilization by microbes, and its stabilization into soil (C associated with soil mineral phase). We also investigated how drying and rewetting influenced N mineralization and loss. We grew wheat (Triticum aestivum) in a controlled environment under constant moisture and under dry-rewetting cycles, and used a continuous ¹³C-labeling method to partition plant and soil organic matter (SOM) contribution to different soil pools. We applied a ¹⁵N label to the soil to determine N loss. We found that dry-rewetting decreased total input of plant C in microbial biomass (MB) and in the soil mineral phase, mainly due to a reduction of plant biomass. Plant derived C in MB and in the soil mineral phase were positively correlated (R² = 0.54; P = 0.0012). N loss was reduced with dry rewetting cycles, and mineralization increased after each rewetting event. Overall drying and rewetting reduced rhizodeposition and stabilization of new C, primary through biomass reduction. However, frequency of rewetting and intensity of drought may determine the fate of C in MB and consequently into the soil mineral phase. Frequency and intensity may also be crucial in stimulating N mineralization and reducing N loss in agricultural soils.     

Distribution of assimilated carbon in the system <Emphasis Type="Italic">Phragmites australis</Emphasis>-waterlogged peat soil after carbon-14 pulse labelling

 Short-term (3–6 days) and long-term (27 days) laboratory experiments were carried out to determine the distribution of assimilated C in the system Phragmites australis (common reed)-waterlogged fen soil after ¹⁴C pulse labelling. The investigated system of fen plants and anaerobic organic soil showed different patterns of assimilated ¹⁴C distribution when compared to systems with cultivated plants and aerobic mineral soil. Between 90% and 95% of the ¹⁴C in the system was found in the reed plants. A maximum of 2% of the assimilated plant ¹⁴C was released from the fen soil as CO₂ and about 5–9% remained in the soil. The ¹⁴C remaining in the waterlogged fen soil of the reed plant had the same amount as that of a cultivated plant in mineral soil, despite lower ¹⁴C-release (i.e. rhizodeposition and root respiration) from reed roots. Assuming that root respiration of fen plants is low, this indicates that microbial C turnover in waterlogged fen soil is much slower than in mineral soil. The estimated quantity of the assimilated C remaining in the soil was of an ecologically relevant order of magnitude. Received: 8 July 1999