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.     

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.     

Distribution and fate of 13C-labeled root and straw residues from ryegrass and crimson clover in soil under western Oregon field conditions

Annual ryegrass (Lolium multiflorum Lam.) and crimson clover (Trifolium incarnatum L.) were pulse-labeled with ¹³C-CO₂ in the field between the initiation of late winter growth (mid-February) and through flowering and seed formation (late May). Straw was harvested after seed maturation (July), and soil containing ¹³C-labeled roots and root-derived C was left in the field until September. ¹³C-enriched and ¹³C-unenriched straw residues of each species were mixed in factorial combinations with soil containing either ¹³C-enriched or ¹³C-unenriched root-derived C and incubated in the field for 10 months. The contributions of C derived from straw, roots, and soil were measured in soil microbial biomass C, respired C, and soil C on five occasions after residue incorporation (September, October, November, April, and June). At straw incorporation (September), 25–30% of soil microbial biomass C was derived from root C in both ryegrass and clover treatments, and this value was sustained in the ryegrass treatment from September to April but declined in the clover treatment. By October, between 20 and 30% of soil microbial biomass C was derived from straw, with the percentage contribution from clover straw generally exceeding that from ryegrass straw throughout the incubation. By June, ryegrass root-derived C contributed 5.5% of the soil C pool, which was significantly greater than the contributions from any of the three other residue types (about 1.5%). This work has provided a framework for more studies of finer scale that should focus on the interactions between residue quality, soil organic matter C, and specific members of the soil microbial community.     

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.     

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.     

Variations in soil microbial biomass and crop roots due to differing resource quality inputs in a tropical dryland agroecosystem

The influence of exogenous organic inputs on soil microbial biomass dynamics and crop root biomass was studied through two annual cycles in rice-barley rotation in a tropical dryland agroecosystem. The treatments involved addition of equivalent amount of N (80 kg N ha⁻¹) through chemical fertilizer and three organic inputs at the beginning of each annual cycle: Sesbania shoot (high-quality resource, C:N 16, lignin:N 3.2, polyphenol+lignin:N 4.2), wheat straw (low-quality resource, C:N 82, lignin:N 34.8, polyphenol+lignin:N 36.8) and Sesbania+wheat straw (high-and low-quality resources combined), besides control. The decomposition rates of various inputs and crop roots were determined in field conditions by mass loss method. Sesbania (decay constant, k=0.028) decomposed much faster than wheat straw (k=0.0025); decomposition rate of Sesbania+wheat straw was twice as fast compared to wheat straw. On average, soil microbial biomass levels were: rice period, Sesbania?Sesbania+wheat straw>wheat straw?fertilizer; barley period, Sesbania+wheat straw>Sesbania?wheat straw?fertilizer; summer fallow, Sesbania+wheat straw>Sesbania>wheat straw?fertilizer. Soil microbial biomass increased through rice and barley crop periods to summer fallow; however, in Sesbania shoot application a strong peak was obtained during rice crop period. In both crops soil microbial biomass C and N decreased distinctly from seedling to grain-forming stages, and then increased to the maximum at crop maturity. Crop roots, however, showed reverse trend through the cropping period, suggesting strong competition between microbial biomass and crop roots for available nutrients. It is concluded that both resource quality and crop roots had distinct effect on soil microbial biomass and combined application of Sesbania shoot and wheat straw was most effective in sustained build up of microbial biomass through the annual cycle.     

Effects of mycorrhizal roots and extraradical hyphae on N uptake from vineyard cover crop litter and the soil microbial community

The objectives of this study were to evaluate the contribution of arbuscular mycorrhizal (AM) fungal hyphae to ¹⁵N uptake from vineyard cover crop litter (Medicago polymorpha), and to examine the soil microbial community under the influence of mycorrhizal roots and extraradical hyphae. Mycorrhizal grapevines (Vitis vinifera) were grown in specially designed containers, within which a polyvinyl chloride (PVC) mesh core was inserted. Different sizes of mesh allowed mycorrhizal roots (mycorrhizosphere treatment) or extraradical hyphae (hyphosphere treatment) to access dual labeled ¹⁵N and ¹³C cover crop litter that was placed inside the cores after 4 months of grapevine growth. Mesh cores in the bulk soil treatment, which served as a negative control, had the same mesh size as the hyphosphere treatment, but frequent rotation prevented extraradical hyphae from accessing the litter. Grapevines and soils were harvested 0, 7, 14, and 28 days after addition of the cover crop litter and examined for the presence of ¹⁵N. Soil microbial biomass and the soil microbial community inside the mesh cores were examined using phospholipid fatty acid analysis. ¹⁵N concentrations in grapevines in the hyphosphere treatment were twice that of grapevines in the bulk soil treatment, suggesting that extraradical hyphae extending from mycorrhizal grapevine roots may have a role in nutrient utilization from decomposing vineyard cover crops in the field. Nonetheless, grapevines in the mycorrhizosphere treatment had the highest ¹⁵N concentrations, thus highlighting the importance of a healthy grapevine root system in nutrient uptake. We detected similar peaks in soil microbial biomass in the mycorrhizosphere and hyphosphere treatments after addition of the litter, despite significantly lower microbial biomass in the hyphosphere treatment initially. Our results suggest that although grapevine roots play a dominant role in the uptake of nutrients from a decomposing cover crop, AM hyphae may have a more important role in maintaining soil microbial communities associated with nutrient cycling.     

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

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

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

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

Carbon flow from C-labeled straw and root residues into the phospholipid fatty acids of a soil microbial community under field conditions

To better understand how residue quality and seasonal conditions influence the flow of C from both root and straw residues into the soil microbial community, we followed the incorporation of ¹³C-labeled crimson clover (Trifolium incarnatum) and ryegrass (Lolium multiflorum) root and straw residues into the phospholipid fatty acids (PLFA) of soil microbial biomass. After residue incorporation under field conditions in late summer (September), the ¹³C content of soil PLFA was measured in September, October, and November, 2002, and April and June, 2003. Multivariate non-metric multidimensional scaling techniques showed that the distribution of ¹³C among microbial PLFA differed among the four primary treatments (ryegrass straw and roots, clover straw and roots). Regardless of treatment, some PLFA remained poorly labeled with ¹³C throughout much of the study (16:1ω5, 10Me17:0; 0-5%), whereas other PLFA consistently contained a larger percentage of residue-derived C (16:0; 18:1ω9, 18:2ω6,9; 10-25%). The distribution of residue ¹³C among individual PLFA differed from the relative contributions of individual PLFA (mol%) to total PLFA-C, suggesting that a subset of the soil biomass was primarily responsible for assimilating residue-derived C. The distribution of ¹³C among soil PLFA differed between the sampling times, indicating that residue properties and soil conditions influenced which members of the community were assimilating residue-derived C. Our findings will provide the foundation for further studies to identify the nature of the community members responsible for residue decomposition at different times of the year, and what factors account for the dynamics of the community involved.     

Carbon sequestration in soils with annual inputs of maize biomass and maize-derived animal manure: Evidence from C abundance

The abundance of ¹³C was determined over a period of nine years in two soils (LUN, coarse sand; ASK, sandy loam) following their conversion from C3-crops and to the C4-crop silage maize (Zea mays L.). The soils were exposed to identical management and climatic conditions, and were sampled every second year. The aboveground maize biomass was either removed (stubbles and roots left), chopped and added to the soil, or fed to sheep and the faeces then added to the soil. Annual inputs of maize biomass and sheep faeces were similar (0.8 kg DM m⁻²). The study included soils maintained under C3-crops (beet roots, Beta vulgaris L.). After nine years of maize cropping, soil C from stubbles and roots accounted for 12 and 16% of the total-C in the LUN and ASK soil, respectively. Without additional organic amendment the content of total-C in the ASK soil remained constant and similar to that of soil retained under C3-crops whereas total-C tended to decrease in the LUN soil. When maize biomass and sheep faeces were added, soil total-C increased and C from these C4-sources averaged 14% and 21% of the soil total-C, respectively. Following nine annual additions, retention of C added in aboveground maize biomass averaged 19% while the retention of C added in maize-derived faeces was 30%. Our study infers that that ruminant manure C contributes about 50% more to soil C sequestration than C applied in crop residues.     

Carbon flow in the plant-soil-microbe continuum at different growth stages of maize grown in a Mollisol

Understanding the photosynthetic carbon (C) dynamics in the plant–soil–microbe continuum is critical to the C sequestration in soils. However, such information is limited in maize (Zea mays L.) in Mollisols. Pot-grown maize was labelled with ¹³CO₂ at the 10-leaf, 15-leaf, heading, milk and dent stages to investigate the photosynthetic C flow in a maize–soil system and its contribution to soil organic carbon (SOC) in Mollisols. The majority of fixed ¹³C was recovered in shoots, ranging from 44.7% to 78.6%. The allocation of ¹³C fixed at different growth stages to belowground (roots and soil) gradually decreased over the growing period, indicating that the strength of root C sink is stronger at the early stages. However, the proportion of ¹³C in dissolved organic C and microbial biomass C to that in SOC significantly increased as the growth stages advanced. Over the entire growth period, the contribution of root-derived C to SOC was estimated to be 5461 mg C plant^?1 growth period^?1, of which approximately 79% was synthesized during the vegetative stages. Therefore, the input of photosynthetic C by maize plants into SOC mainly occurred during the younger stages of the plant, favouring the storage of SOC in Mollisols.     

Natural abundance δN and δC of DNA extracted from soil

We report the first simultaneous measurements of δ¹⁵N and δ¹³C of DNA extracted from surface soils. The isotopic composition of DNA differed significantly among nine different soils. The δ¹³C and δ¹⁵N of DNA was correlated with δ¹³C and δ¹⁵N of soil, respectively, suggesting that the isotopic composition of DNA is strongly influenced by the isotopic composition of soil organic matter. However, in all samples DNA was enriched in ¹³C relative to soil, indicating microorganisms fractionated C during assimilation or preferentially used ¹³C enriched substrates. Enrichment of DNA in ¹⁵N relative to soil was not consistently observed, but there were significant differences between δ¹⁵N of DNA and δ¹⁵N of soil for three different sites, suggesting microorganisms are fractionating N or preferentially using N substrates at different rates across these contrasting ecosystems. There was a strong linear correlation between δ¹⁵N of DNA and δ¹⁵N of the microbial biomass, which indicated DNA was depleted in ¹⁵N relative to the microbial biomass by approximately 3.4‰. Our results show that accurate and precise isotopic measurements of C and N in DNA extracted from the soil are feasible, and that these analyses may provide powerful tools for elucidating C and N cycling processes through soil microorganisms.     

Modification of the original chloroform fumigation extraction technique to allow measurement of δC of soil microbial biomass carbon

In studies of the soil microbial biomass C by the chloroform fumigation extraction (CFE) technique, biomass C is routinely extracted using 0.5 M K₂SO₄ solution. The excessive amounts of salts contained in the extracting solution pause a significant challenge in using ¹³C isotope techniques to study the nature of C in the soil microbial biomass. This is because the salts can affect the oxidation process and therefore hamper accurate mass spectromic analysis of dried extracts. In spite of this, no standard protocol exists for preparing the K₂SO₄ extracts for ¹³C isotope analysis. We have modified the original CFE method to allow measurement of the δ¹³C of soil microbial biomass C by using 2 M KCl instead of the usual 0.5 M K₂SO₄ solution to extract biomass C. Excess salts were removed by dialysis in 100 molecular weight cut off membranes, after which the extracts were freeze-dried and their δ¹³C measured using a mass spectrometer. The soil microbial biomass C and δ¹³C of 2 M KCl extracts were compared with those of 0.5 M K₂SO₄ extracts. There was excellent agreement between organic C and δ¹³C estimates for dialyzed 2 M KCl and 0.5 M K₂SO₄ extracts, but the speed of dialysis for the latter was very slow, making use of the former more rapid. These results suggest that in procedures where oxidation with potassium dichromate is not critical to analysis of soluble C, 2 M KCl may be used in place of 0.5 M K₂SO₄ to extract soil microbial biomass C for δ¹³C measurements. The new procedure is relatively easy and rapid for obtaining indices for both pool sizes and turnover rates of soil microbial biomass C and provides a promising approach to study soil organic C.     

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.     

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

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

不同施肥处理对光合碳在花生-土壤系统中分配的影响

通过室内花生盆栽,设置NPK(常规氮磷钾施肥)、NPKS(常规氮磷钾加玉米秸秆)、NPKA(常规氮磷钾加腐殖酸)和CK(不施肥对照)4个不同的施肥处理,采用3次~(13)CO2脉冲标记的方法对不同施肥处理下光合碳在花生-土壤系统中的分配进行定量研究。结果表明:不同施肥处理对标记期内花生总生物量影响不显著,但是NPKA处理显著提升了花生根系生物量,较CK、NPK和NPKS分别高22.04%、19.47%和53.38%。NPKS处理地上部~(13)C丰度最高,但土壤中~(13)C丰度最低,NPKA处理土壤中~(13)C丰度最高。各处理地上部的~(13)C含量无显著差异,NPKA处理根系的~(13)C含量显著高于NPK且土壤~(13)C含量显著高于其他处理。NPKA处理地上部的~(13)C分配比例最低而土壤中分配比例最高,根系~(13)C分配比例与其他处理无显著差异,根系与土壤~(13)C分配比例之和显著高于其他处理。本研究表明腐殖酸能显著促进花生光合碳向地下部的转运。     

Microbial community composition and carbon cycling within soil microenvironments of conventional, low-input, and organic cropping systems

This study coupled stable isotope probing with phospholipid fatty acid analysis (¹³C-PLFA) to describe the role of microbial community composition in the short-term processing (i.e., C incorporation into microbial biomass and/or deposition or respiration of C) of root- versus residue-C and, ultimately, in long-term C sequestration in conventional (annual synthetic fertilizer applications), low-input (synthetic fertilizer and cover crop applied in alternating years), and organic (annual composted manure and cover crop additions) maize-tomato (Zea mays - Lycopersicum esculentum) cropping systems. During the maize growing season, we traced ¹³C-labeled hairy vetch (Vicia dasycarpa) roots and residues into PLFAs extracted from soil microaggregates (53-250 μm) and silt-and-clay (<53 μm) particles. Total PLFA biomass was greatest in the organic (41.4 nmol g⁻¹ soil) and similar between the conventional and low-input systems (31.0 and 30.1 nmol g⁻¹ soil, respectively), with Gram-positive bacterial PLFA dominating the microbial communities in all systems. Although total PLFA-C derived from roots was over four times greater than from residues, relative distributions (mol%) of root- and residue-derived C into the microbial communities were not different among the three cropping systems. Additionally, neither the PLFA profiles nor the amount of root- and residue-C incorporation into the PLFAs of the microaggregates were consistently different when compared with the silt-and-clay particles. More fungal PLFA-C was measured, however, in microaggregates compared with silt-and-clay. The lack of differences between the mol% within the microbial communities of the cropping systems and between the PLFA-C in the microaggregates and the silt-and-clay may have been due to (i) insufficient differences in quality between roots and residues and/or (ii) the high N availability in these N-fertilized cropping systems that augmented the abilities of the microbial communities to process a wide range of substrate qualities. The main implications of this study are that (i) the greater short-term microbial processing of root- than residue-C can be a mechanistic explanation for the higher relative retention of root- over residue-C, but microbial community composition did not influence long-term C sequestration trends in the three cropping systems and (ii) in spite of the similarity between the microbial community profiles of the microaggregates and the silt-and-clay, more C was processed in the microaggregates by fungi, suggesting that the microaggregate is a relatively unique microenvironment for fungal activity.     

Compound-specific δC and δH analyses of plant and soil organic matter: A preliminary assessment of the effects of vegetation change on ecosystem hydrology

Here we present δ¹³C and δ²H data of long-chained, even-numbered (C₂₇-C₃₁) n-alkanes from C3 (trees) and C4 (grasses) plants and from the corresponding soils from a grassland-woodland vegetation sequence in central Queensland, Australia. Our data show that δ¹³C values of the C4 grassland species were heavier relative to those of C3 tree species from the woodland (Acacia leaves) and woody grassland (Atalaya leaves). However, n-alkanes from the C4 grasses had lighter δ²H values relative to the Acacia leaves, but showed no significant difference in δ²H values when compared with C3 Atalaya leaves. These results differ from those of previous studies, showing that C4 grasses had heavier δ²H values relative to C3 grasses and trees. Those observations have been explained by C4 plants accessing the more evaporation-influenced and isotopically heavier surface water and tree roots sourcing deeper, isotopically lighter soil water (“Two-layered soil-water system”). By comparison, our data suggest that ecosystem changes (vegetation “thickening”) can significantly alter the soil hydrological characteristics. This is shown by the heavier δ²H values in the woodland soil compared with lighter δ²H values in the grassland soil, implying that the recent vegetation change (increased tree biomass) in the woodland had altered soil hydrological conditions. Estimated δ²H values of the source-water for vegetation in the grassland and woodland showed that both trees and grasses in open settings accessed water with lighter δ²H values (avg. −46‰) compared with water accessed by trees in the woodland vegetation (avg. −7‰). These data suggest that in semi-arid environments the “two-layer” soil water concept might not apply. Furthermore, our data indicate that compound-specific δ²H and δ¹³C analyses of n-alkanes from soil organic matter can be used to successfully differentiate between water sources of different vegetation types (grasses versus trees) in natural ecosystems.     

Rhizosphere priming of soil organic matter by bacterial groups in a grassland soil

Plants often impact the rate of native soil organic matter turnover through root interactions with soil organisms; however the role of root-microbial interactions in mediation of the “priming effect” is not well understood. We examined the effects of living plant roots and N fertilization on belowground C dynamics in a California annual grassland soil (Haploxeralf) during a two-year greenhouse study. The fate of ¹³C-labeled belowground C (roots and organic matter) was followed under planted (Avena barbata) and unplanted conditions, and with and without supplemental N (20 kg N ha⁻¹ season⁻¹) over two periods of plant growth, each followed by a dry, fallow period of 120 d. Turnover of belowground ¹³C SOM was followed using ¹³C-phospholipid fatty acid (PLFA) biomarkers. Living roots increased the turnover and loss of belowground ¹³C compared with unplanted soils. Planted soils had 20% less belowground ¹³C present than in unplanted soils after 2 cycles of planting and fallow. After 2 treatment cycles, unlabeled soil C was 4.8% higher in planted soils than unplanted. The addition of N to soils decreased the turnover of enriched belowground ¹³C during the first treatment season in both planted and unplanted soils, however no effect of N was observed thereafter. Our findings suggest that A. barbata may increase soil C levels over time because root and exudate C inputs are significant, but that increase will be moderated by an overall faster C mineralization rate of belowground C. N addition may slow soil C losses; however, the effect was minor and transient in this system. The labeled root-derived ¹³C was initially recovered in gram negative (highest enrichment), gram positive, and fungal biomarkers. With successive growing seasons, the labeled C in the gram negative and fungal markers declined, while gram positive markers continued to accumulate labeled belowground C. The rhizosphere of A. barbata shifted the microbial community composition, resulting in greater abundances of gram negative markers and lower abundances of gram positive, actinobacteria and cyclopropyl PLFA markers compared to unplanted soil. However, the longer-term utilization of labeled belowground C by gram positive bacteria was enhanced in the rhizosphere microbial community compared with unplanted soils. We suggest that the activities of gram positive bacteria may be major controllers of multi-year rhizosphere-related priming of SOM decomposition.