Dynamic study of the burrowing behaviour of Aporrectodea nocturna and Allolobophora chlorotica: interactions between earthworms and spatial avoidance of burrows

The behaviour of earthworms belonging to two different species and ecological types (Aporrectodea nocturna and Allolobophora chlorotica) was studied using two-dimensional (2D) terraria. Two experiments were set up to gain insight into the nature of interactions between these earthworms. Firstly, the evolution of the burrow systems was analysed with the density of the earthworms varying from one to five individuals. Secondly, a burrow system was first established by using one earthworm which was then removed before the introduction of a second earthworm. This second earthworm therefore encountered a burrow system created either by a conspecific earthworm or by an earthworm of the other species. These experiments showed that: (1) intra- and interspecific interactions occur between earthworms, (2) these interactions are dependent on the physical presence of the earthworms, and (3) spatial avoidance can occur (A. chlorotica avoiding burrows created by A. nocturna). The results suggest that earthworm burrow systems are "individual structures", rarely used by other earthworms when inhabited. When abandoned, the burrows may be recolonised depending on the ecological type of the earthworm under consideration.     

Lethal and sublethal effects of imidacloprid on two earthworm species (Aporrectodea nocturna and Allolobophora icterica)

The chemical imidacloprid is the major component of many widely used insecticides and is relatively persistent in soils. A set of experiments was carried out to estimate the lethal (mortality) and sublethal (weight loss) effects of one of these insecticides, Confidor, on two earthworm species commonly found in agricultural soils. A preliminary experiment in the absence of earthworms showed that imidacloprid was not rapidly degraded, with a decrease of less than 10% after 2 weeks, and that it was distributed in a reasonably homogeneous manner throughout the soil (less than 10% of variation between samples). The LC₅₀ of imidacloprid for the anecic species Aporrectodea nocturna and the endogeic species Allolobophora icterica was between 2 and 4 mg kg^–1 dry soil. This result is consistent with previous findings obtained with other earthworm species and natural soils. When sublethal effects were examined, significant decreases in weight were observed at concentrations of 0.5 and 1 mg kg^–1 dry soil for the two earthworm species whereas no effect was observed at a concentration of 0.1 mg kg^–1 dry soil (NOEC value). These concentrations are close to 0.33 mg kg^–1 which is the Predictive Environmental Concentration. Weight loss appears to be a valuable endpoint that can be used with worms freshly collected in the field as long as variability in the response of a control is taken into account.     

Sublethal effects of imidacloprid on the burrowing behaviour of two earthworm species: Modifications of the 3D burrow systems in artificial cores and consequences on gas diffusion in soil

Earthworms have been termed ‘ecosystem engineers’ (sensu [Jones, C.G., Lawton, J.H., Shachak, M., 1994. Organisms as ecosysem engineers. Oikos 69, 373-386.]) because of the important roles they play in the soil. As a consequence, it is assumed that if earthworms change their behaviour following exposure to pesticides or pollutants this could have a drastic impact on soil functioning. To test this assumption under laboratory conditions, we studied the burrow systems made by two earthworm species (the anecic Aporrectodea nocturna and the endogeic Allolobophora icterica) in artificial soil cores containing imidacloprid, a widely used neonicotinoid insecticide. After 1-month incubation period, the macropores created in the soil core were analyzed by tomography. In order to further characterize transfer properties associated with burrow systems gas diffusion measurements were also carried out. The burrow systems made by the two earthworm species were very different: A. nocturna made more continuous, less branched, more vertical and wider burrows than A. icterica. Some changes to A. nocturna burrow systems were observed after exposure to imidacloprid (they made a smaller burrow system and burrows were more narrow), but only at the highest concentration of imidacloprid used (0.5 mg kg⁻¹). A. icterica worms were more sensitive to imidacloprid and many differences in their burrow systems (length, sinuosity, branching rate and number of burrows) were observed at both concentrations tested (0.1 and 0. 5 mg kg⁻¹). As a consequence, the continuity of the burrow systems made by both species was altered following imidacloprid treatment. Gas diffusion through the A. nocturna soil cores was reduced but no difference in gas diffusion was observed in the A. icterica soil cores.     

Colonization behaviour of two enterobacterial strains on cereals

 Two diazotrophic enterobacterial strains, Pantoea agglomerans D5/23 and Klebsiella pneumoniae CC12/12, were observed in sterile and non-sterile hydroponic and soil experiments in order to determine, by means of an immunological detection method (double antibody sandwich enzyme linked immunosorbent assay), their colonization sites, their migration within individuals of different plant species, and their ability to compete with indigenous organisms. To investigate the interaction between bacteria and plants, root and shoot samples were analysed using electron microscopy. Field experiments were performed to determine the growth-promoting abilities of the bacterial strains. In field experiments, inoculation with P. agglomerans led to an increase in the grain yield of different wheat (Triticum aestivum) cultivars. The same strain was also able to colonize the rhizosphere and the phyllosphere of different cereals due to its ability to migrate within the plant. Roots and media were colonized 10–100 times more intensively than shoots, with about 10⁶ cells g^–1 root and 10⁴ cells g^–1 shoot. We found that P. agglomerans colonized the root and plant-growth medium of wheat to a greater extent than those of rye (Secale cereale) and barley (Hordeum vulgare), whereas the colonization of shoots was higher in rye and barley compared to wheat. Furthermore, while cell numbers of K. pneumoniae in media and roots were 10 times higher than cell numbers of P. agglomerans, only the latter markedly increased root growth. We were able to detect significant differences in colonization numbers between treatments even if the data were not normally or log-normally distributed or the variances were not homogenous. Received: 14 June 1999     

Evolution of burrow systems after the accidental introduction of a new earthworm species into a Swiss pre-alpine meadow

 The unintentional introduction of a new earthworm species (Aporrectodea nocturna) into a Swiss pre-alpine meadow resulted in a great increase in earthworm density in the newly colonized area (386 m^–2) compared with the density observed in the natural area (273 m^–2) where an earthworm community was already present. To investigate the impact of this introduction on the burrow systems, eight soil cores (length 25 cm, diameter 16 cm) were taken (four in the colonized area and four in the natural area) and analysed with computer-assisted X-ray tomography. The resulting images were processed to obtain the 3D-skeleton reconstructions of the earthworm burrow systems. Due to high variability in these burrow systems, only slight differences were observed between the two areas. The total burrow length and the mean burrow lengths tended to be greater in the colonized area. Moreover, the distribution of pore numbers with depth showed different patterns with a maximum for depths between 10 cm and 15 cm in the colonized area and a maximum for depths between 20 cm and 25 cm in the natural area. These differences may have been related to: (1) the particular behaviour of A. nocturna, which was observed to cast at the surface in this site, and (2) the predominance of juvenile earthworms around the colonization front. These differences were sufficient to create significant effects on the continuity of the burrow systems (assessed by the number of different pathways between virtual horizontal planes) for the two areas. The colonized area was characterized by a greater pore continuity, which could have resulted in enhanced transfer properties. Received: 2 July 1999     

Seasonal variations in growth and reproduction of the earthworm Perionyx excavatus (Oligochaeta: Megascolecidae)

 One-week-old juveniles of Perionyx excavatus were cultured in urine-free cow manure for 16 weeks in three different seasons to find out the influence of environmental factors on growth and reproduction. The biomass of worms increased gradually until the 16th week in all seasons. There was no significant (F=2.51) variation in biomass between the three seasons; however, the growth rate varied significantly (F=50.49, P<0.01). During the pre-clitellar period, the growth rate (mg/g) increased steeply (1013.17±41.33 in the monsoon season, 728.64±15.5 in winter and 463.16±22.96 in summer) until the worms attained maturity (5–8 weeks) and from then onwards gradually decreased after the commencement of cocoon production. Worms attained maturity when the mean weight of each worm ranged from 167.46 mg to 197.13 mg. The cumulative number of cocoons varied significantly (F=26.07, P<0.01) with season. Similarly, the rate of cocoon production also varied significantly (F=62.06, P<0.01). The average rate of cocoon production was 7.23, 0.99 and 0.53/worm/week in the monsoon season, winter and summer, respectively. By week 16, the ratio of weight gain to the initial weight of 1-week-old juveniles was 1 : 43.13 in the monsoon season, 1 : 30.49 in the winter and 1 : 23.69 in the summer. The maximum mass gain, growth rate, and cocoon production and earliest attainment of maturity were recorded during the monsoon season, and were attributed to the lowest range of fluctuating temperatures and high humidity compared with during winter and summer. The growth rate of worms in all seasons was inversely proportional to the number of cocoons produced. Further, the higher the rate of cocoon production, the lower the weight of individual cocoons. Received: 1 July 1997     

Characterization of the burrow system of the earthworms Lumbricus terrestris and Aporrectodea giardi using X-ray computed tomography and image analysis

 The burrow systems of two earthworm species (Lumbricus terrestris and Aporrectodea giardi) were studied in artificially packed soil columns placed in controlled conditions in the laboratory. At the end of the incubation, which lasted 246 days, the burrow systems were characterized on the undisturbed columns using X-ray computed tomography. This method provided a set of digitized images corresponding to horizontal 3-mm-thick sections. The following parameters were measured using image analysis on each section: number of biopores, their volume, and their individual orientation calculated using an elliptical model of the earthworm channel. The profiles of these parameters through the columns showed that the burrow systems of L. terrestris and A. giardi, which are both anecic species, were very different in terms of total volume, number of burrows, burrow orientation and extension with soil depth. These results led us to conclude that the burrow system of L. terrestris can be considered as a permanent structure whereas that of A. giardi is closer to the burrow system of endogeic species. Received: 12 June 1998     

Effects of trefoil cover crop and earthworm inoculation on maize crop and soil organisms in Reunion Island

 Traditional tree fallows have been abandoned on the western coast of the Reunion Island because of the increasing need for cultivated land. Soil fertility is no longer restored and crop yields have decreased drastically. The leguminous plant, Lotus uliginosus (trefoil), used as a cover crop, has made possible the control of erosion, the restoration of soil macrofauna, especially earthworms, and the increase in crop yields. When trefoil was associated with earthworms (Amynthas corticis), the densities of maize, the yields of maize stalk and dry matter, the yield of trefoil fodder dry matter, and the biomass and respiratory activity of soil microflora were considerably increased. The combined effects of their association led to a significant decrease in populations of the plant-parasitic nematode, Pratylenchus vulnus, in maize roots, and in the population of borers. Some soil chemical features were modified. Received: 10 September 1997     

Carbon and nitrogen dynamics in ageing earthworm casts in grasslands of the eastern plains of Colombia

 The effects of a large species of anecic earthworm, Martiodrilus carimaguensis Jiménez and Moreno, on soil C and N dynamics were investigated in a native savanna and a man-made pasture of the eastern plains of Colombia. We compared, across time (11 months), the total C, total N, NH⁺ ₄ and NO^– ₃ contents in the earthworm casts, the underlying soil and the adjacent soil. Additional sampling of root biomass and macrofauna was performed. In the two management systems, the total C and N contents were higher in casts (4.33–7.50%) than in the bulk soil (2.81–4.08%), showing that the earthworms selected food substrates with high organic contents. In general, C contents significantly increased during cast ageing (+100%), possibly because of CO₂ fixation processes, dead root accumulation and/or macrofaunal activities in casts. In fresh casts, NH⁺ ₄ levels were very high (294.20–233.98 μg g^–1 dry cast) when compared to the soil (26.96–73.95 μg g^–1 dry soil), due to the intense mineralisation processes that occurred during the transit of soil and organic matter through the earthworm gut. During the first week of cast ageing, NH⁺ ₄ levels sharply decreased, while NH^– ₃ levels showed successive peaks in the casts, the underlying soil and the adjacent soil. These results suggested the rapid production of NO^– ₃ by nitrification processes in the fresh casts, followed by diffusion to the nearby soil, first vertically, then horizontally. After 2 weeks of cast ageing, NH⁺ ₄ and NO^– ₃ levels only showed slight variations, likely because of organic matter protection in stable dry casts. The root biomass was higher (1.6–4.7 times) below the old earthworm casts. The ecological significance of these results is discussed. Received: 22 October 1998     

Growth and cocoon production by the earthworm Metaphire houletti (Oligochaeta) in different food sources

 Quantitative observations on the biology of Metaphire houletti were made in cow and horse manure and oak litter in laboratory conditions over a period of 240 days. The study revealed that copulation is not a prerequisite for production of viable cocoons, indicating that M. houletti may be parthenogenetic. The mean growth rate was 2.86 mg worm^–1 day^–1 reaching sexual maturity at 45 days and producing 0.015 cocoons worm^–1 day^–1 (kept singly) and 2.82 mg worm^–1 day^–1, reaching sexual maturity at 45 days and producing 0.03 cocoons worm^–1 day^–1 (kept in batches) in cow manure; 4.08 mg worm^–1 day^–1, reaching sexual maturity at 46 days and producing 0.02 cocoons worm^–1 day^–1 (kept singly) and 2.97 mg worm^–1 day^–1, reaching sexual maturity at 45 days and producing 0.016 cocoons worm^–1 day^–1 (kept in batches) in horse manure; 3.73 mg worm^–1 day^–1 reaching sexual maturity at 45 day and producing 0.023 cocoons worm^–1 day^–1 (kept singly) and 2.73 mg worm^–1 day^–1, reaching sexual maturity at 47 days and producing 0.028 cocoons worm^–1 day^–1 (kept in batches) in oak litter. After an incubation period of 31.9±1.2 days 82% of the cocoons hatched with a mean of 1.12±0.06 hatchlings per cocoon. The earthworms reared in batches did not demonstrate any advantage over those reared singly in all substrates. Higher growth rates were observed in earthworms raised singly than those raised in batches in all substrates. Received: 30 April 1998     

Accumulation of zinc and its effects on the growth, reproduction and life cycle of Drawida willsi (Oligochaeta), a dominant earthworm in Indian crop fields

 Accumulation of Zn and its effects on the growth, reproduction and life cycle of the earthworm Drawida willsi were determined. D. willsi did not reveal any significant changes in their mass at any of the concentrations of Zn (50, 200 and 400 mg kg^–1) compared to in untreated soils. The Zn concentrations in the exposed earthworms were significantly increased, but they were able to regulate their body content of Zn within a range of 116–125 mg kg^–1 (dry wt) in 200–400 mg kg^–1 Zn-treated soil. Reproduction was significantly reduced when the Zn concentration in soil exceeded 200 mg kg^–1. The drop in reproduction at elevated concentrations of Zn apparently resulted in a delay in completion of the life cycle and a decline in the total population. Received: 9 September 1998     

Temporal and spatial fluctuations in moisture affect humus microfungal community structure in microcosms

 The effects of temporal and spatial fluctuations in moisture on the microfungal community were studied in birch litter plus pine humus microcosms over 6 months. Two treatments were used: a uniform treatment in which moisture was maintained at the original moisture content throughout the profile over the course of the experiment; and a fluctuating moisture treatment in which the moisture content of the whole microcosm fluctuated weekly around the original moisture content. At 6 months, fungal species richness was higher and diversity was lower under moisture fluctuations than under uniform moisture. The number of fungal isolates and proportional diversity was significantly higher in the bottom layer in the fluctuating moisture treatment. The fluctuating moisture treatment and the bottom layer correlated significantly with the first PCA axis, which accounted for 66% of the variation in the fungal species data. The experiment suggests that even without drying, fluctuations in moisture can affect fungal community structure. Received: 3 June 1999     

The effects of low lead levels on the growth and reproduction of the African earthworm Eudrilus eugeniae (Oligochaeta)

 The effects of sublethal concentrations of lead nitrate on the growth and reproduction of the African composting earthworm species, Eudrilus eugeniae, was studied by exposing worms in an organic substrate to lead-nitrate-contaminated food over a period of 76 days. The results revealed that growth was initially affected negatively by the presence of lead, while the maturation rate and cocoon production were not affected. In agreement with other studies on Eisenia fetida, cocoon viability was affected negatively by lead, making this a sensitive toxicity endpoint. Received: 14 October 1998     

Assessment of recovery of population, biomass and reproduction of the earthworm Drawida willsi following the application of malathion under field conditions

 The recovery of population, biomass and reproduction of a dominant crop field earthworm (Drawida willsi, Michaelsen) was assessed following the application of two recommended agricultural doses of malathion (2.2 mg kg^–1– single dose and 4.4 mg kg^–1– double dose) in a rice field agroecosystem for a period of 105 days. The average worm population and biomass showed 12% and 21% decline, respectively, in plots treated with single and double doses of malathion, compared with control. The peak of worm population was observed after 60 days in control (plot I) and this peak value was shifted by 15 days (i.e. observed after 75 days) in malathion-treated plots (plots II and III) indicating delay in growth and maturation of the worms following the application of malathion. Inhibition in the total number of cocoons produced, and thus in the rate of reproduction, was also observed in malathion-treated plots. The peak rate of reproduction (1.71 cocoons adult^–1) was observed after 60 days in control plots and after 75 days (1.57 and 1.40 cocoons adult^–1) in single and double dose malathion-treated plots, respectively. One-way analysis of variance showed significant differences in the population size and biomass of D. willsi up to 75 days, in rate of reproduction up to 90 days, and no difference thereafter. This indicates that, under field conditions, D. willsi worms took about 75–90 days after application of malathion to resume normal population, biomass and reproduction, and hence we suggest that a second application of malathion in single and double agricultural doses should be avoided before this time. Received: 7 September 1999     

High performance liquid chromatography studies on the polysaccharides in the walls of the mounds of two species of termite in Senegal,Cubitermes oculatus and Macrotermes subhyalinus: their origin and contribution to structural stability

 The origin, nature and quantity of polysaccharides in the walls of the epigeal mounds of a species of soil-feeding termite, Cubitermes oculatus, and a fungus-growing termite, Macrotermes subhyalinus, found in Senegal, and of soil not considered to be under the influence of termites, were studied to obtain a clearer picture of the structural stability of these materials. The compounds were extractedand analysed by high performance liquid chromatography. We found that the walls of mounds made by soil-feeding species were very rich in sugars soluble in aqueous acid or hot water. Most of the sugars originated from cellulose and hemicellulose, and only a small proportion from microorganisms. There were also significant amounts of stachyose in the mound walls and in the reference soil. This sugar was probably formed by the surrounding vegetation, which was mainly leguminous crops. Comparison of the mineral and organic-mineral particle sizes of samples confirmed that the walls of soil-feeding termite mounds where there is the greatest redistribution of clay have the best aggregating capacity. The results therefore show that the polysaccharides in mound walls of soil-feeding termites are mostly of plant origin. Their influence on the stability of these structure is discussed. The walls of fungus-growing termite mounds contain little organic matter and hence low levels of polysaccharides, which are mainly of plant origin. Received: 19 July 1999     

Survival and cell culturability of biocontrol Pseudomonas fluorescens CHA0 in the rhizosphere of cucumber grown in two soils of contrasting fertility status

 The effect of cucumber roots on survival patterns of the biocontrol soil inoculant Pseudomonas fluorescens CHA0-Rif was assessed for 22 days in two non-sterile soils, using a combination of total immunofluorescence cell counts, Kogure's direct viable counts and colony counts on plates containing rifampicin. In Eschikon soil (high fertility status for cucumber), CHA0-Rif persisted as culturable cells in bulk soil and in the rhizosphere, but colony counts were lower than viable counts and total cell counts inside root tissues. The occurrence of viable but non-culturable (VBNC) cells inside root tissues (5 log cells g^–1 root) was unlikely to have resulted from the hydrogen peroxide treatment used to disinfect the root surface, as hydrogen peroxide caused the death of CHA0-Rif cells in vitro. In Siglistorf soil (low fertility status for cucumber), the inoculant was found mostly as non-culturable cells. Colony counts and viable counts of CHA0-Rif were similar, both in bulk soil and inside root tissues, whereas in the rhizosphere viable counts exceeded colony counts at the last two samplings (giving about 7 log VBNC cells g^–1). In conclusion, soil type had a significant influence on the occurrence of VBNC cells of CHA0-Rif, although these cells were found in root-associated habitats (i.e. rhizosphere and root tissues) and not in bulk soil. Received: 12 November 1999     

Influence of stock camping behaviour on the soil microbiological and biochemical properties of grazed pastoral soils

 The size and activity of the soil microbial biomass in grazed pastures was compared on the main grazing area and on stock camp areas where animals congregate. Two sites were on hill country and three on gently sloping border-dyke irrigated land. Due to the transfer of nutrients and organic matter to the camp areas via dung and urine there was an accumulation of soil organic C, organic and inorganic P and S and soluble salts in the camp areas. Soil pH also tended to be higher in camp areas due to transfer of alkalinity by the grazing animals. Water soluble organic C, microbial biomass C and basal respiration were all higher in soils from camp areas but the proportion of organic C present as microbial C and the microbial respiratory quotient were unaffected. Microbial activity as quantified by arginine ammonification rate and fluorescein diacetate (FDA) hydrolysis was higher in camp than non-camp soils but dehydrogenase activity remained unaffected. Activities of protease, histidase, urease, acid phosphatase and aryl-sulphatase were all higher in stock camp soils. The activities of both histidase and aryl-sulphatase were also higher when expressed per unit of microbial biomass C, indicating that the increased activity was the result of increased enzyme production by the microbial community. Prolonged regular applications of dairy shed effluent (diluted dung and urine from cattle) to a field had a similar effect to stock camping in increasing soil organic matter content, nutrient accumulation and soil biological activity. It was concluded that the stock camping activity of grazing animals results in an increase in both the fertility and biological activity in soils from camp areas at the expense of these properties on the main grazing areas. Received: 20 October 1997     

Relationships between woodlice (Isopoda: Oniscidea) and microbial density and activity in the field

 Field studies on microbial populations and their activity in soil and litter layers of different sites revealed statistically significant correlations with climatic factors, soil characteristics and the number of terrestrial isopods (Isopoda: Oniscidea). The number of captured Porcellio scaber was negatively correlated with the density of soil microbiota, but positively correlated with both respiration and cellulolytic activity of litter-colonizing microorganisms. By contrast, the number of Oniscus asellus was positively correlated with soil microbial density and respiration, and cellulolytic and respiratory activity of the litter. These interspecific differences were unexpected between species that frequently occur sympatrically, and, thus, probably have similar ecological requirements. Hence, the results suggested species-specific interactions of these species with soil and litter-colonizing microbiota. However, further field studies are required to prove this. Received: 16 December 1998     

Effect of salt stress on interaction between lentil (Lens culinaris) genotypes and Rhizobium spp. strains: symbiotic N2 fixation in normal and sodic soils

 Following screening, selection, characterization and examination of their symbiotic N₂ fixation, only two Rhizobium strains (ND-16 and TAL-1860) and four lentil genotypes (DLG-103, LC-50, LC-53 and Sehore 74-3) were found to be suited to sodic soils. Interactions between salt-tolerant lentil genotypes and Rhizobium strains were found to be significant, and resulted in greater nodulation, N₂ fixation (nitrogenase activity), total nitrogen, plant height, root length and grain yield in sodic soils under field conditions compared to uninoculated controls. Significantly more nodulation, nitrogenase activity, glutamine synthetase (GS) and NADH-dependent glutamate synthase (NADH-GOGAT) activities were found in normal soil as compared to the soil supplemented with 4% and 8% NaCl. Salt stress inhibited nitrogenase, GS and NADH-GOGAT activities. However, nitrogenase activity in nodules was more sensitive to salt stress than GS and NADH-GOGAT activities (NH₄ ⁺ assimilation). The relevance of these findings for salt-tolerant symbionts is discussed. Received: 14 November 1997     

Nodulation and nitrogen fixation of Lupinus species with Bradyrhizobium (lupin) strains in iron-deficient soil

 The effect of six Bradyrhizobium sp. (lupin) strains (WPBS 3201D, WPBS 3211D, USDA 3040, USDA 3041, USDA 3042 and CB 2272) and Fe supply on nodulation, N₂-fixation and growth of three lupin species (Lupinus termis, L. albus and L. triticale) grown under Fe deficiency in an alkaline soil, were examined in sterilized and non-sterilized pot experiments. When inoculated with USDA 3040, 3041, 3042 and CB2272 without Fe addition, the three lupin species had a very low nodule number and mass, low shoot and root dry matter accumulation and lower N yield. However, inoculation with WPBS 3201D and 3211D without Fe treatments increased all these parameters substantially. The ability of WPBS 3201D and 3211D to form nodules on the three lupin species under conditions of Fe stress could be attributed to their ability to scavenge Fe from Fe-deficient environments through their siderophore production. Addition of Fe to the other four strains significantly increased nodulation and N₂-fixation of the three lupin species, indicating that the poorer nodulation and N₂-fixation of these strains in the absence of Fe, resulted from a low ability to obtain Fe from alkaline soils. Bradyrhizobium strains WPBS 3201D and 3211D were superior to the other four strains in terms of promoting greater nodulation, N₂-fixation, plant growth and N accumulation of L. termis and L. albus. However, the other four strains were more efficient in symbiotic association with L. triticale. The greater variations in nodule efficiencies (specific nitrogenase activity) under different levels of Fe supply could be attributed to the quantities of bacteroid protein and leghaemoglobin in the nodules. The results suggested that Bradyrhizobium (lupin) strains differ greatly in their ability to obtain Fe from alkaline soils, and that the selection of bradyrhizobial strains which are tolerant of Fe deficient soils could complement plant breeding for the selection of legume crops for Fe-deficient soils. Received: 5 January 1998