Plant availability of phosphorus in superphosphate and a phosphate rock as influenced by earthworms

The effect of earthworms on the plant availability of phosphorus (P) in superphosphate and Chatham Rise phosphorite (CRP) was evaluated in a glasshouse experiment using perennial ryegrass over seven harvests. A mixed earthworm population of Lumbricus rubellus (Hoff.) and Allolobophora caliginosa (Savigny) was used. Increases in the yield of ryegrass in the presence of earthworms varied from 2 to 32%, whereas increases in P uptake by ryegrass ranged from 0 to 40% over seven harvests. With superphosphate, the initial increases in both ryegrass yield and P uptake by ryegrass in the presence of earthworms ranged from 20 to 40% at first harvest to less than 10% by the seventh. In marked contrast, earthworms increased the agronomic performance of pelletized CRP by 15 to 30% throughout the trial period. An increase in plant-available soil N concentrations due to earthworm activity probably explains the initial difference in the performance of superphosphate. The increased agronomic effectiveness of CRP appears to result from the incorporation and intimate mixing of the PR with the soil by earthworms. The implications of the results obtained in the present study to the interpretation of glasshouse and field trials evaluating P fertilizers are also discussed.     

Stable isotope N and C labelling of different functional groups of earthworms and their casts: A tool for studying trophic links

Earthworms (Oligochaeta: Lumbricidae) have substantial effects on the structure and fertility of soils with consequences for the diversity of plant communities and associated ecosystem functions. However, we still lack a clear understanding of the functional role earthworms play in terrestrial ecosystems, partly because easy-to-use methods to quantify their activities are missing. In this study, we tested whether earthworms and their casts can be dual-labelled with ¹⁵N and ¹³C stable isotopes by cultivating them in soil substrate amended with ¹⁵N ammonium nitrate and ¹³C-glucose. Additionally, we also wanted to know whether (i) earthworms from different functional groups (soil-feeders vs. litter-feeders) and their casts would differ in their incorporation of stable isotopes, (ii) if enrichment levels are higher if the same amount of isotopes is applied in one dose or in staggered doses, and (iii) if isotopic enrichment in casts changes when they are stored in a conditioning cabinet or in a pot filled with soil placed in a greenhouse. Our findings show the feasibility of dual-labelling tissues and casts of both litter-feeding (Lumbricus terrestris) and soil-feeding (Aporrectodea caliginosa) earthworms using the same method. The advantage of this method is that earthworms and their casts can be labelled under realistic conditions by cultivating them for only four days in soil that received a one-time addition of commercially available stable isotopes instead of offering labelled plant material. In earthworms, the isotopic enrichment remained at a stable level for at least 21 days; labelled casts could be stored for at least 105 days without significantly decreasing their isotopic signals. This simple and efficient method opens new avenues for studying the role of these important ecosystem engineers in nutrient cycling and their functional relationships with other organisms.     

Land Application of Aluminum Water Treatment Residual: Aluminum Phytoavailability and Forage Yield

Negative impacts of land‐applied aluminum (Al)–rich water treatment residuals (WTRs), which have been suggested to improve soil phosphorus (P) sorption, could include excessive immobilization of plant‐available P and Al phytotoxicity. We studied the impacts of an Al‐rich WTRs on agronomic returns and plant Al concentrations in glasshouse and field studies. The glasshouse study was a 4 × 2 × 3 factorial experiment with one control in a randomized complete block design and three replicates. Four sources of P were each applied at two agronomic rate [44 kg P ha^?1, P‐based rate; and 179 kg plant‐available nitrogen (PAN) ha^?1, N‐based rate] to topsoil (0–15 cm) of a sandy, siliceous, hyperthermic Arenic Alaquods. Three WTR rates (0, 10, and 25 g kg^?1 oven‐dry‐weight basis) were further applied, whereas the control received neither P source nor WTRs. Bahiagrass (Paspalum notatum Fluggae), ryegrass (Lolium perenne L.), and a second bahiagrass crop were continuously grown in succession for 18 months. Applied WTRs increased soil Al but not plant Al concentrations (22–80 mg Al kg^?1), which fell within the normal concentration range for pasture plants. In the glasshouse, when WTRs were incorporated with the soil, bahiagrass dry matter (DM) accumulation was reduced, but ryegrass DM was not affected even at 25 g kg^?1 WTR. A 2‐year field study, with same treatments but two rates of WTRs (0 and 10 g kg^?1 WTR) surface applied to established bahiagrass on the same soil type (Spodosols) showed neither reduced yields nor increased plant Al phytoavailability in the WTR treatments. The studies show no increase in plant Al is associated with Al‐WTRs applied to reduce excess soil‐soluble P and P losses but plant DM accumulation may be reduced.     

Mucus excretion and carbon turnover of endogeic earthworms

Summary Mucus excretion of endogeic earthworms, by the body surface to burrow walls and by the intestine to casts, was investigated using uniformly ¹⁴C-labelled adult or subadult specimens of Octolasion lacteum (Örley) in laboratory incubations in soil from a beechwood on limestone. The daily loss of C due to mucus excretion from the body surface and in casts was calculated as 0.2 and 0.5% of total animal C, respectively. The C loss due to mucus excretion by subadult or adult individuals of O. lacteum is assumed to account for 63% of total C losses (including mucus excretion and respiration) of the earthworms. In a second experiment we studied the incorporation of ¹⁴C from labelled soil, again from a beechwood on limestone, into the tissue of the endogeic earthworm species Aporrectodea caliginosa (Savigny). The results of this experiment indicate the existence of two C pools, one more labile and one more stable, in earthworms. It is assumed that the C investment for respiration and mucus excretion is derived from the labile C pool of endogeic earthworms.     

蚯蚓与黑麦草相互作用对土壤中荧蒽去除的影响

Earthworms can promote the bioremediation of contaminated soils through enhancing plant growth and microorganism development. The individual and combined effects of earthworms and ryegrass (Lolium multiflorum Lam.) on the removal of fluoranthene from a sandy-loam alluvial soil were investigated in a 70-d microcosm experiment. The experiment was set up in a complete factorial design with treatments in four replicates: without earthworms or ryegrass (control, CK), with earthworms only (E), with ryegrass only (P), and with both earthworms and ryegrass (EP). The residual fluoranthene, microbial biomass C, and polyphenol oxidase activity in the soil changed significantly (P<0.01) with time. In general, the residual concentration of fluoranthene in the soil decreased sharply from 71.8-88.7 to 31.7-37.4 mg kg 1 in 14 d, and then decreased gradually to 19.7-30.5 mg kg 1 on the 70th d. The fluoranthene concentration left in the soil was the least with both earthworms and ryegrass, compared to the other treatments at the end of the experiment. Half-life times of fluoranthene in the E, P, and EP treatments were 17.8%-36.3% smaller than that of CK. More fluoranthene was absorbed by earthworms than ryegrass. However, the total amounts of fluoranthene accumulated in both the ryegrass and earthworms were small, only accounting for 0.01%-1.20% of the lost fluoranthene. Therefore, we assumed that microbial degradation would play a dominant functional role in fluoranthene removal from soil. We found that earthworms significantly increased microbial biomass C and polyphenol oxidase activity (P<0.01) in the presence of ryegrass at the end of the experiment. Furthermore, microbial biomass C and polyphenol oxidase activity were significantly (P<0.05) and negatively related to the residual fluoranthene concentration. This implied that earthworms might promote the removal of fluoranthene from soil via stimulating microbial biomass C and polyphenol oxidase activity.     

Magnesium influence on plant uptake of phosphorus in a calcareous soil

A pot experiment was conducted to investigate the role of magnesium (Mg) in plant utilization of ³²phoshorus (P)‐labelled P in a calcareous soil (Typic Hapluquent). Results with two successive harvests of ryegrass shoots indicated that Mg had no obvious influence on plant uptake of the P applied. In a separate incubation study with the same soil, labelled P along with different levels of Mg was introduced to the soil at two time intervals and soil inorganic P was fractionated. The isotope data of P in various fractions indicated that Mg at the test levels had no obvious influence on P transformation either. It is suggested from the experiments that Mg is unlikely to be able to promote plant utilization of fertilizer P in calcareous soils.     

Quantifying dung carbon incorporation by earthworms in pasture soils

The effect of different earthworm functional groups on the incorporation of maize (C4 plant) dung into a soil (C3 organic matter background) sown with ryegrass (C3 plant) was explored by using differences in the carbon (C) isotope ratios (¹²C and ¹³C) between plant and soil samples in a field mesocosm study. The abundance of earthworms increased with dung inputs, reaching over 4000 earthworms per m², presumably because of the increased food resources used. The amount of dung C incorporated into the soil profile in the presence of earthworms was dependent on the amount of organic matter deposited on the soil surface (925–4620 g C m^?2) and reached rates of 1200 g C m^?2 annually in the treatment receiving repeat dung applications. Dung incorporation was largely concentrated in the surface 0–75 mm, although small amounts of dung‐derived C were observed to a depth of 300 mm. This was especially so in the presence of anecic earthworms, equating to an extra 70 g C m^?2 annually for the 150–300 mm depth increment. It is important to note, in calculating C incorporation rates from earthworms, that only 10–20% of the soil surface in grazed pastures is covered by dung. After 444 days, less than 32% of the applied dung was detected within the upper 300 mm of the soil profile. This study emphasized the need for all three earthworm functional groups to be present within the soil in order to maximize the amount of surface dung that could be incorporated into soil organic matter.     

Cycling of nitrogen by surface-casting earthworms in a pasture ecosystem

Production of surface casts and the removal of plant litter from the soil surface by earth-worms had similar seasonal variations, with maximum values in May and minimum values occurring in July and August. Seasonal variations in the total nitrogen (TN) and oxidizable carbon contents of casts were closely related to variations in litter production. The C:N ratio of casts (10.7) was consistently smaller than that of underlying soil material (15.0 and 14.2 for the 0–5 and 18–22 cm depths, respectively), which is probably due to the mineralization of plant-derived organic material during passage through earthworms and utilization of low C:N ratio litter. Seasonal changes in the amounts of inorganic N forms in casts showed a build-up of NH₄⁺N in the cooler winter months (July and August), attaining a maximum of 112 μg.g^?1. with a decrease in autumn (April and May) and early spring (September and October), reaching a minimum of 54 μg.g^?1. The opposite trend existed for seasonal variations in the NO₃^?-N content of the casts. Because only minor fluctuations in the amounts of N forms were obtained for underlying soil material during the casting period, the more dramatic changes observed in the casts could not be explained by soil variations. Seasonal variations in urease enzyme activity, associated with fluctuations in organic matter content, were more important than the effect of temperature on enzyme activity in accounting for seasonal variations in the NH₄⁺-N content of casts. It was calculated that 73% of the TN content of litter removed from the surface by earthworms was accumulated in casts, indicating both the importance of earthworms in incorporating litter N with soil material and the inefficiency of N digestion by earthworms.     

Subsurface earthworm casts can be important soil microsites specifically influencing the growth of grassland plants

Earthworms (Annelida: Oligochaeta) deposit several tons per hectare of casts enriched in nutrients and/or arbuscular mycorrhizal fungi (AMF) and create a spatial and temporal soil heterogeneity that can play a role in structuring plant communities. However, while we begin to understand the role of surface casts, it is still unclear to what extent plants utilize subsurface casts. We conducted a greenhouse experiment using large mesocosms (volume 45 l) to test whether (1) soil microsites consisting of earthworm casts with or without AMF (four Glomus taxa) affect the biomass production of 11 grassland plant species comprising the three functional groups grasses, forbs, and legumes, (2) different ecological groups of earthworms (soil dwellers—Aporrectodea caliginosa vs. vertical burrowers—Lumbricus terrestris) alter potential influences of soil microsites (i.e., four earthworms × two subsurface microsites × two AMF treatments). Soil microsites were artificially inserted in a 25-cm depth, and afterwards, plant species were sown in a regular pattern; the experiment ran for 6 months. Our results show that minute amounts of subsurface casts (0.89 g kg^?1 soil) decreased the shoot and root production of forbs and legumes, but not that of grasses. The presence of earthworms reduced root biomass of grasses only. Our data also suggest that subsurface casts provide microsites from which root AMF colonization can start. Ecological groups of earthworms did not differ in their effects on plant production or AMF distribution. Taken together, these findings suggest that subsurface earthworm casts might play a role in structuring plant communities by specifically affecting the growth of certain functional groups of plants.     

Decomposition of 15N-labelled catch-crop residues in soil: evaluation of N mineralization and plant-N uptake potentials under controlled conditions

The decomposition of ¹⁵N-labelled catch-crop materials (rape, radish and rye), obtained from field experiments, was studied in a chalky Champagne soil during a 60-week incubation at 28°C. Mineralized N was assumed to come from either labile or recalcitrant fractions of plant residues. The labile fraction represented about one-third of the catch-crop N; its mineralization rate constant varied from 0.06 to 0.12 d^?1. The decomposition rate of the recalcitrant N fraction ranged from 0.03 × 10^?2 to 0.06 × 10^?2 d^?1. Catch-crop species and rate of incorporation had no effect on N residue mineralized at the end of incubation. The decomposition of labelled rye was monitored in the same soil during a 5-month pot experiment to determine the N availability to an Italian ryegrass crop and the effect of plants on the decomposition processes. The ¹⁵N-rye decomposed rapidly both in the presence or absence of Italian ryegrass, but the amounts of N mineralized were influenced by the presence of living roots: 42% of the ¹⁵N in labelled rye was present as inorganic N in the pots without plants after 5 months, compared with only 32% in the ryegrass crop. Comparison of microbial-biomass dynamics in both treatments suggested that there had been preferential utilization by soil micro-organisms of materials released from the living roots than the labelled plant residues.     

Effects of earthworms and ryegrass on respiratory and enzyme activities of soil

The effects of earthworms (Allolobophora caliginosa (Savigny)) and ryegrass (Lolium perenne L.) on the respiratory and enzyme activities of the subsoil of Judgeford silt loam, from which 15 cm of topsoil had been removed, were studied in a glasshouse pot trial over some 13 months. Treatments were subsoil alone, subsoil + ryegrass, subsoil + earthworms, and subsoil + earthworms + ryegrass; activities were assessed every 4 months.In subsoil alone, oxygen uptakes and xylanase, urease, phosphatase, and sulphatase activities declined, but invertase activity increased, during the trial. The presence of earthworms resulted in increased oxygen uptakes, and cellulase and sulphatase activities at some samplings.Generally, the presence of ryegrass enhanced all of the biochemical activities. The additional presence of earthworms further stimulated the activities of invertase, amylase, urease, and phosphatase.Earthworms therefore stimulate biochemical activities and nutrient cycling, and thereby contribute to the restoration of pasture productivity after topsoil removal.     

Origin of the effect of earthworms on the availability of phosphorus in a phosphate rock

A series of trials was undertaken to evaluate the mechanisms by which earthworms increase the availability of phosphorus (P) in a phosphate rock (PR). In the 6 days that PR pellets were visible on the soil surface, there was no downward movement from the site of application in the presence of earthworms. Results from the glasshouse study with tillage treatments also suggest that earthworms play no direct role in the incorporation of the surface-applied PR. Addition of earthworms (Lumbricus rubellus (Hoff.)) to the soil incubating with SPR resulted in a 32% increase in Bray-extractable soil P after 70 days and increases ranging from 30 to 44% in bicarbonate-extractable soil P over the same period. Dissolution of PR, measured by extraction with 0.5 m NaOH, also increased in the presence of earthworms after 70 days from 24% (120 μg P) to 32% (160 μgP) of added P. These increases in dissolution of the PR in the incubating soil in the presence of earthworms appear to result largely from an increase in the degree of intimate contact of the PR particles with soil surfaces. This was confirmed by optical microscopy. Whereas estimates of ingestion rates, obtained by two traditional methods, indicated that L. rubellus ingested less than 5% of the incubating soil over 70 days, measurement of the proportion of soil affected by earthworms on an area basis using a frame technique ranged from 66% for Allolobophora caliginosa (Savigny) and 89% for L. rubellus, to 100% for the two species combined after 19 days. Results obtained with the frame technique explain more fully the effect of earthworms on the dissolution of the PR during incubation.     

蚓粪对黑麦草吸收污染土壤重金属铜的影响

在长江冲积物形成的高沙土中加入CuSO4.5H2O模拟土壤污染,使Cu污染浓度分别为200、400、600 mg kg-1,并设置加蚓粪(15%)和加原土(15%)处理,二处理各设种植黑麦草(Lolium multiflorum)和不种黑麦草培养试验,研究蚓粪对黑麦草生长及对Cu吸收的影响,以揭示蚓粪在Cu污染土壤植物修复中的作用。结果表明:蚓粪显著增加了黑麦草的地上部和地下部的生物量(p<0.001**)以及根系的长度、表面积、体积和根尖数(p<0.05*),在Cu浓度为200 mg kg-1时促进作用最大;蚓粪还显著提高了黑麦草地上部Cu的浓度及累积量,但显著降低了地下部的Cu浓度(p<0.05*),而对地下部的铜累积量没有影响,显示蚓粪能促进Cu从根系向地上部的运移及在地上部的富集。通过对土壤的pH和Cu的形态特征分析,发现种植黑麦草处理降低了土壤的pH并增加了可交换态铜的含量,而且这种作用在加入蚓粪后更加明显。推测蚓粪主要通过促进黑麦草根系的生长和活性而影响根系周围环境,提高重金属铜的生物有效性进而增加植物对铜的吸收。     

Stable isotope labelling of earthworms can help deciphering belowground–aboveground interactions involving earthworms,mycorrhizal fungi,plants and aphids

Functional relationships between belowground detritivores and/or symbionts and aboveground primary producers and their herbivores are not well studied. In a factorial greenhouse experiment we studied interactions between earthworms (addition/no addition of Lumbricus terrestris; Clitellata: Lumbricidae) and arbuscular-mycorrhizal fungi (AMF; with/without inoculation of Glomus mosseae; Glomerales: Glomeraceae) on the leguminous herb Trifolium repens (Fabales: Fabaceae) and associated plant aphids (Aphis gossypii, A. craccivora; Hemiptera: Aphidoidea). In order to be able to trace organismic interactions, earthworms were dual-labelled with stable isotopes (¹⁵N-ammonium nitrate and ¹³C-glucose). We specifically wanted to investigate whether (i) isotopic signals can be traced from the labelled earthworms via surface castings, plant roots and leaves to plant aphids and (ii) these compartments differ in their incorporation of stable isotopes. Our results show that the tested organismic compartments differed significantly in their ¹⁵N isotope enrichments measured seven days after the introduction of earthworms. ¹⁵N isotope incorporation was highest in casts followed by earthworm tissue, roots and leaves, with lowest ¹⁵N signature in aphids. The ¹³C signal in roots, leaves and aphids was similar across all treatments and is for this reason not recommendable for tracing short-term interactions over multitrophic levels. AMF symbiosis affected stable isotope incorporation differently in different subsystems: the ¹⁵N isotope signature was higher below ground (in roots) but lower above ground (leaves and aphids) in AMF-inoculated mesocosms compared to AMF-free mesocosms (significant subsystem × AMF interaction). Aphid infestation was unaffected by AMF and/or earthworms. Generally, these results demonstrate that plants utilize nutrients excreted by earthworms and incorporate these nutrients into their roots, leaf tissue and phloem sap from where aphids suck. Hence, these results show that earthworms and plant aphids are functionally interlinked. Further, ¹⁵N-labelling earthworms may represent a promising tool to investigate nutrient uptake by plants and consequences for belowground-aboveground multitrophic interactions.     

Phosphorus fractions and dynamics in surface earthworm casts under native and improved grasslands in a Colombian savanna Oxisol

The objective of this study was to assess the effect of a native anecic species on phosphorus availability in an Oxisol characterised by a low chemical fertility. Experiments were carried out at Carimagua research station in a representative site of the isohyperthermic savannas on the Colombian Orinoco basin. One field study and two laboratory/incubation studies were performed in a natural herbaceous savanna and a Brachiaria decumbens and Pueraria phaseoloides pasture. In the laboratory, experiment pots were prepared containing soil collected from the respective field paddock's topsoil. Total P content was higher in earthworm casts than in the surrounding soil in field samples, 50% in native savanna soil and more than 100% in pasture soil. In casts produced under laboratory conditions this increase was relatively low (10-20%). Under field conditions, almost without exception, all P fractions were increased in casts relative to the original soil (corresponding to the increase in total P content), being relatively greater in the labile inorganic P fractions. In addition, samples from the natural savanna showed that pH of casts was higher (5.2) than that of soil (4.6) in both field and laboratory samples. Except in the native savanna under field conditions, the phosphatase activity was reduced in casts by 16.7 to 44%. From our results we conclude that earthworms in the field incorporate P from litter or other organic sources (i.e. undecomposed plant and root material, earthworm faeces) which is not normally measured in the analysis of bulk soil.     

Earthworms of different functional groups affect the fate of the Bt-toxin Cry1Ab from transgenic maize in soil

The fate of the insecticidal Cry1Ab protein from crop residues (leaves and roots) of the transgenic maize variety MON810 was studied in the presence and absence of two earthworm species (Lumbricus terrestris, Aporrectodea caliginosa; separate incubations) in soil microcosms. The recombinant Cry1Ab protein was quantified using a highly sensitive ELISA. Control microcosms received corresponding non-transgenic plant material. All earthworms survived in the microcosms over a period of 5 weeks, irrespective of whether they received MON810 or non-transgenic plant material. Weight loss was observed for both earthworm species, independent of the plant material or transgenic modification. A strong decline of immunoreactive Cry1Ab in plant residues (mean initial concentration approx. 5000 ng g⁻¹) of MON810 was observed in all treatments, but in microcosms with earthworms this decline was significantly higher with less than 10% of the initial Cry1Ab concentration remaining after 5 weeks. Cry1Ab concentrations in casts were only 0.1% of those found in remaining plant material of the respective microcosms. No immunoreactive Cry1Ab proteins were found in earthworm tissues (threshold of detection: 0.58 ng g⁻¹ fresh weight). No further decline was found for Cry1Ab concentrations in casts of A. caliginosa during a subsequent period of 3 months of incubation in bulk soil (<0.1 ng g⁻¹) after removal of the earthworms from the microcosms, while in casts of L. terrestris the concentration decreased from 0.4 to below 0.1 ng g⁻¹. In conclusion, this study demonstrates that earthworms enhance the decline of immunoreactive Cry1Ab proteins from maize residues.     

Relative Phosphorus Phytoavailability of Different Phosphorus Sources

Understanding differences in the phytoavailability of various phosphorus (P) sources should improve matching P additions to plant needs and minimize excessive buildup of bioavailable P, which can degrade aquatic systems. We evaluated relative P phytoavailability (RPP) of different P sources in glasshouse and field studies. Bahiagrass (Paspalum notatum Fluggae), ryegrass (Lolium perenne L.), and a second bahiagrass crop were grown in succession in a P‐deficient soil amended with four sources of P (triple superphosphate (TSP), Boca Raton and Pompano; biosolids, and poultry manure), each applied at two rates in the glasshouse study and to an established bahiagrass pasture in the field. The RPP values estimated from plant P uptake of each organic source of P relative to TSP in the glasshouse were similar for the three croppings and similar to the estimates derived from the field study, but varied for the different P sources. Values ranged from 30% for poultry manure to 85% for Boca Raton biosolids. Boca Raton biosolid P was as readily available as P in TSP and would be classified as a high RPP (>75% RPP) source, but Pompano biosolids and manure would be classified as moderate RPP materials (25–75% RPP). The RPP values observed in manure and Pompano biosolid treatments are consistent with 50% “effectiveness” suggested for biosolid P in U.S. Environmental Protection Agency (USEPA) guidelines, whereas P bioavailability of Boca Raton biosolids is similar to mineral fertilizer.     

Earthworms disseminate a soil-borne plant pathogen,Fusarium oxysporum f. sp. raphani

Radish plants infested with a soil-borne plant pathogen, Fusarium oxysporum f. sp. raphani PEG-4, which is resistant to hygromycin B, were placed on the surface of a soil microcosm containing earthworms (Pheretima sp.). The earthworms ate the radish plants and scattered individual casts everywhere in the burrows. The fungal propagules were detected in the gut of the earthworms and in 26 out of 28 casts. These results suggested that the pathogen could survive passage through the alimentary canal of the earthworms and be disseminated anywhere earthworms wandered. The collected casts were incubated in a moist chamber at 28°C for 14 days, which resulted in a decline in the number of the propagules. The effects of the earthworms on the population dynamics of the pathogen were estimated. The earthworms seemed to cause a decline in total propagules of the pathogen in soil, although they expanded its distribution in soil.     

Rapid incorporation of carbon from fresh residues into newly formed stable microaggregates within earthworm casts

Earthworms play an important role in protecting carbon in the soil, but the exact influence of their activity on the distribution and protection of C is still poorly understood. We investigated the effect of earthworms on the formation of stable microaggregates inside newly formed macroaggregates and the distribution of C in them. We crushed (< 250 µm) soil, and subjected it to three treatments: (i) soil + ¹³C‐labelled residue + earthworms (these added after 8 days' incubation), (ii) soil + ¹³C‐labelled residue, and (iii) control (no additions), and then incubated it for 20 days. At the end, we measured the aggregate size distribution, total C and ¹³C, and we isolated microaggregates (53–250 µm) from macroaggregates (> 250 µm) formed. The ¹³C in fine particulate organic matter between and within the microaggregates was determined. Earthworms helped to form large macroaggregates (> 2000 µm). These large macroaggregates contained four times more stable microaggregates than those from samples without earthworms. There was more particulate organic matter within and between microaggregates in macroaggregates in the presence of earthworms. The larger amounts of organic matter inside stable microaggregates in casts than in bulk soil after 12 days of incubation (140 mg ¹³C kg^?1 soil compared with 20 mg ¹³C kg^?1 soil) indicates that these microaggregates are formed rapidly around freshly incorporated residues within casts. In conclusion, earthworms have a direct impact on the formation of stable microaggregates and the incorporation of organic matter inside these microaggregates, and it seems likely that their activity is of great significance for the long‐term stabilization of organic matter in soils.     

STUDIES ON THE DECOMPOSITION OF PLANT MATERIAL IN SOIL. V. THE EFFECTS OF PLANT COVER AND SOIL TYPE ON THE LOSS OF CARBON FROM14C LABELLED RYEGRASS DECOMPOSING UNDER FIELD CONDITIONS

Ryegrass uniformly labelled with ^{1 4}C was allowed to decompose for 10 years under field conditions in a range of contrasting soils. The amount of organic matter already in a soil had no effect on the retention of labelled C by that soil, nor had a variation in soil pH of from 4.9 to 8.1. Decomposition was initially slower in a strongly acid soil (pH 3.7) but by the end of 5 years the difference between this soil and the others had almost disappeared. The more clay in a soil, the greater the retention of labelled C over the whole 10 year period; this was true of both strongly acid and near-neutral soils. More labelled organic matter was leached from a soil containing 7.6% clay than from one with 17.5% clay, but the amount thus lost was insufficient to account for the difference in retention of C by the two soils. The decomposition of labelled plant material was faster in bare soil than in soil growing grass but the ‘protection’ thus given to the labelled C by the growing grass ended when the grass was removed. In bare soil about one third of the labelled ryegrass C was left after one year but thereafter decomposition became very much slower and about one eighth of the labelled C still remained in the soil after 10 years. The decay curve can be represented by a two compartment model, in which about 70% of the ryegrass C decomposed by a first order process of half life 0.25 years and the remainder by a similar process of half-life 8 years.