Hydrolytic enzyme activities in maize (Zea mays) and sorghum (Sorghum bicolor) roots inoculated with Gluconacetobacter diazotrophicus and Glomus intraradices

Two strains of Gluconacetobacter diazotrophicus (Pal 5, UAP5541) and the arbuscular mycorrhizal fungus Glomus intraradices increased both the shoot and root dry weight of sorghum 45 days after inoculation, whereas they had no effect on the shoot and root dry weight of maize. Co-inoculation (Gluconacetobacter diazotrophicus plus Glomus mosseae) did not increase the shoot and root dry weight of either plant. There was a synergistic effect of Gluconacetobacter diazotrophicus on root colonization of maize by Glomus intraradices, whereas an antagonistic interaction was observed in the sorghum root where the number of Gluconacetobacter diazotrophicus and the colonization by Glomus intraradices were reduced. Plant roots inoculated with Gluconacetobacter diazotrophicus and Glomus intraradices, either separately or together, significantly increased root endoglucanase, endopolymethylgalacturonase and endoxyloglucanase activities. The increase varied according to the plant. For example, in comparison with non-inoculated plants, there were higher endoglucanase (+328%), endopolymethylgalacturonase (+180%) and endoxyloglucanase (+125%) activities in 45-day old co-inoculated maize, but not in 45-day old sorghum. The possibility is discussed that hydrolytic enzyme activities were increased as a result of inoculation with Gluconacetobacter diazotrophicus, considering this to be one of the mechanisms by which these bacteria may increase root colonization by AM fungi.     

Nickel-tolerant Brevibacillus brevis and arbuscular mycorrhizal fungus can reduce metal acquisition and nickel toxicity effects in plant growing in nickel supplemented soil

The growth of clover (Trifolium repens ) and its uptake of N, P and Ni were studied following inoculation of soil with Rhizobium trifolii, and combinations of two Ni-adapted indigenous bacterial isolates (one of them was Brevibacillus brevis) and an arbuscular mycorrhizal (AM) fungus (Glomus mosseae). Plant growth was measured in a pot experiment containing soil spiked with 30 (Ni I), 90 (Ni II) or 270 (Ni III) mg kg⁻¹ Ni-sulphate (corresponding to 11.7, 27.6 and 65.8 mg kg⁻¹ available Ni on a dry soil basis). Single inoculation with the most Ni-tolerant bacterial isolate (Brevibacillus brevis) was particularly effective in increasing shoot and root biomass at the three levels of Ni contamination in comparison with the other indigenous bacterial inoculated or control plants. Single colonisation of G. mosseae enhanced by 3 fold (Ni I), by 2.4 fold (Ni II) and by 2.2 fold (Ni III) T. repens dry weight and P-content of the shoots increased by 9.8 fold (Ni I), by 9.9 fold (Ni II) and by 5.1 fold (Ni III) concomitantly with a reduction in Ni concentration in the shoot compared with non-treated plants. Coinoculation of G. mosseae and the Ni-tolerant bacterial strain (B. brevis) achieved the highest plant dry biomass (shoot and root) and N and P content and the lowest Ni shoot concentration. Dual inoculation with the most Ni-tolerant autochthonous microorganisms (B. brevis and G. mosseae) increased shoot and root plant biomass and subtantially reduced the specific absorption rate (defined as the amount of metal absorbed per unit of root biomass) for nickel in comparison with plants grown in soil inoculated only with G. mosseae. B. brevis increased nodule number that was highly depressed in Ni I added soil or supressed in Ni II and Ni III supplemented soil. These results suggest that selected bacterial inoculation improved the mycorrhizal benefit in nutrients uptake and in decreasing Ni toxicity. Inoculation of adapted beneficial microorganisms (as autochthonous B. brevis and G. mosseae) may be used as a tool to enhance plant performance in soil contaminated with Ni.     

Interaction between the soil yeast Rhodotorula mucilaginosa and the arbuscular mycorrhizal fungi Glomus mosseae and Gigaspora rosea

The effect of the soil yeast, Rhodotorula mucilaginosa LBA, on Glomus mosseae (BEG n°12) and Gigaspora rosea (BEG n°9) was studied in vitro and in greenhouse trials. Hyphal length of G. mosseae and G. rosea spores increased significantly in the presence of R. mucilaginosa. Exudates from R. mucilaginosa stimulated hyphal growth of G. mosseae and G. rosea spores. Increase in hyphal length of G. mosseae coincided with an increase in R. mucilaginosa exudates. No stimulation of G. rosea hyphal growth was detected when 0.3 and 0.5 ml per petri dish of yeast exudates was applied. Percentage root length colonization by G. mosseae in soybean (Glycine max L. Merill) and by G. rosea in red clover (Trifolium pratense L. cv. Huia) was increased only when the soil yeast was inoculated before G. mosseae or G. rosea was introduced. Beneficial effects of R. mucilaginosa on arbuscular mycorrhizal (AM) colonization were found when the soil yeast was inoculated either as a thin agar slice or as a volume of 5 and 10 ml of an aqueous solution. R. mucilaginosa exudates (20 ml per pots) applied to soil increased significantly the percentage of AM colonization of soybean and red clover.     

Physiological characteristics (SDH and ALP activities) of arbuscular mycorrhizal colonization as affected by Bacillus thuringiensis inoculation under two phosphorus levels

The effect of Bacillus thuringiensis (B.t.) inoculation on plant growth and on the intra- and extraradical mycorrhizal development of lettuce roots colonized by Glomus mosseae or Glomus intraradices was examined in an inert, soil-less substrate. Histochemical determination of succinate dehydrogenase (SDH) and alkaline phosphatase (ALP) activities which indicate active fungal metabolism was carried out at two phosphorus (P) levels. The presence of B.t. increased extra- and intraradical colonization [measured as frequency (%F), intensity (%I) and percentage of arbuscules (%A)] for both arbuscular mycorrhizal fungi (AMF) rather than plant growth or nutrition regardless P level. Under the lowest level of P fertilization, B.t. enhanced to a similar extent the extra- and intraradical development of both endophytes, but the proportion of fungal tissue showing SDH or ALP was increased in G. intraradices-colonized plants. [SDH: 458% (M) and 512% (A); ALP: 358% (M) and 300% (A)]. P supply decreased G. intraradices colonization to a higher extent than G. mosseae. Nevertheless, the totality of G. intraradices structures developed in P-amended medium showed intraradical o extraradical activity, while in G. mosseae-colonized roots, SDH and ALP activities highly decreased relative to fungal tissue determined by TB staining as affected by P. Our results show that bacterial inoculation compensates the negative effect of P on the intraradical fungal growth and vitality. P amendment reduced in a higher extent G. intraradices infection intensity (non-vital and vital staining) and G. mosseae activity (ALP staining). Thus, big differences in the proportion of SDH-active infection showing ALP activity in mycelium developed by each endophyte were noted at the highest P level. Physiological plant parameters such as photosynthetic activity did not explain specific changes on each arbuscular-mycorrhizal fungus as affected by P or B.t. inoculation. The increased extraradical mycelium development and metabolic fungal activity as a result of B.t. inoculation positively affected N and P plant content and photosynthetic rate in G. intraradices-colonized plants under the lowest P conditions. In general, the increased metabolically active fungal biomass in co-inoculated plants was irrespective of P level and was not related to the P plant uptake from the inert soil-less substrate. These results show the bacterial effect increasing the physiological and metabolic status of AM endophytes, which not only confirms but also extends previous findings on arbuscular mycorrhizae-bacteria interactions. The present study emphasizes the ecological and practical importance of rhizosphere free-living bacteria as mycorrhizae-helper microorganisms.     

Interactions between azospirillum and va mycorrhiza and their effects on growth and nutrition of maize and ryegrass

Interactions between the N₂-fixing bacterium Azospirillum brasilense and the mycorrhizal fungus Glomus mosseae were studied in relation to their effects on the growth and nutrition of Zea mays (C₄) and Lolium perenne (C₃) plants. Although roots from plants inoculated with Azospirillum exhibited C₂H₂ reduction activity no significant effect of inoculation on N concentration in the plant shoots was found. With non-mycorrhizal plants, inoculation with Azospirillum resulted in increased dry matter production at the first harvest compared to the effect achieved by supplying N as fertilizer, but this trend was reversed at the last harvest. However, with mycorrhizal maize plants, Azospirillum, which stimulated the development of VA mycorrhiza, was still effective in improving plant growth and nutrient uptake at the last harvest. Azospirillum and N behaved similarly in enhancing the growth and nutrition of mycorrhizal maize. The dual inoculation of maize by Azospirillum and Glomus produced plants of a similar size, N content, and a higher P content, than those supplied with N and P.     

Using arbuscular mycorrhiza to reduce the stressful effects of soil compaction on corn (Zea mays L.) growth

Soil compaction is of great importance in agriculture, because its high levels may adversely affect plant growth and the environment. Since mechanical methods are not very efficient and economical, using biological methods to alleviate the stress of soil compaction on plant growth may be beneficial. The objectives of this study were to: (1) evaluate the effects of soil compaction on corn (Zea mays L.) growth, and (2) test the hypothesis that applying arbuscular mycorrhiza (AM) with different origins can partially or completely overcome the stressful effects of soil compaction on corn growth under unsterilized and sterilized conditions. Corn was planted in unsterilized and sterilized compacted soils, while treated with three species of AM including, Iranian Glomus mosseae, Iranian Glomus etunicatum, and Canadian Glomus mosseae, received from GINCO (Glomales in vitro Collection), Canada. Plant growth variables and soil resistance parameters were determined. AM significantly increased root fresh (maximum of 94% increase) and dry (maximum of 100% increase) weights in the compacted soil. AM with different origins may improve corn growth in compacted soils, though its effectiveness is related to the level of compaction and also to the interaction with other soil microorganisms.     

Interference in culture among Rhizobium strains

Growth interference in culture was shown by 57 strains of Rhizobium belonging to different inoculation groups. The assays were conducted in vitro by spot tests on agar plates and using 1007 paired combinations, 240 cases of interference were observed.A frequency of interference of about 30% was found among strains of R. meliloti and R. meliloti, R. lupini, R. trifolii and R. leguminosarum. Only one case out of 168 showed interference between R. meliloti and R. japonicum.     

Interactions between the arbuscular mycorrhizal fungus Glomus mosseae and plant growth-promoting fungi and their significance for enhancing plant growth and suppressing damping-off of cucumber (Cucumis sativus L.)

This study investigated the interactions between the arbuscular mycorrhizal fungus (AMF) Glomus mosseae and the plant growth-promoting fungi (PGPF) Penicillium simplicissimum GP17-2 and Trichoderma harzianum GT3-2 in relation to their colonization of roots and the rhizosphere of cucumber (Cucumis sativus L.), and their effect on plant growth and suppression of the damping-off pathogen Rhizoctonia solani. Combined inoculation of T. harzianum with G. mosseae increased the percentage of AMF root colonization, but the presence of P. simplicissimum had no significant effect on mycorrhizal formation. The existence of G. mosseae decreased the population development of T. harzianum in and/or around the roots, however, growth of P. simplicissimum was not affected. Both PGPF isolates were effective in increasing the plant shoot and root dry weight, G. mosseae, however, was not. Nevertheless, co-inoculation of G. mosseae with T. harzianum synergistically enhanced plant growth. A comparable additive effect on plant growth was not observed when P. simplicissimum was combined with G. mosseae. Treating plants with the PGPF either simultaneously with the pathogen or prior to pathogen infection suppressed damping-off disease. G. mosseae was significantly effective in reducing disease severity when inoculated prior to the pathogen but not when inoculated simultaneously with the pathogen. Interestingly, the levels of disease protection achieved by the single inoculation of P. simplicissimum, T. harzianum or G. mosseae were significantly increased by combined inoculation of each PGPF with G. mosseae.     

An AM fungus and a PGPR intensify the adverse effects of salinity on the stability of rhizosphere soil aggregates of Lactuca sativa

A mesocosm experiment was conducted to examine the effect of an arbuscular mycorrhizal (AM) fungus (Glomus mosseae (Nicol & Gerd.) Gerd. & Trappe) and a plant growth-promoting rhizobacterium (PGPR) (Pseudomonas mendocina Palleroni), alone or in combination, on the structural stability of the rhizosphere soil of Lactuca sativa L. grown under two levels of salinity. The plants inoculated with P. mendocina had significantly greater shoot biomass than the control plants at both salinity levels, whereas the mycorrhizal inoculation was only effective in increasing shoot biomass at the moderate salinity level. The aggregate stability of soils inoculated with the PGPR and/or G. mosseae significantly decreased with increasing saline stress (about 29% lower than those of soils under non-saline conditions). Only the inoculated soils showed higher concentrations of sodium (Na) under severe saline stress. The severe salinity stress decreased the glomalin-related soil protein (GRSP) concentration, but the highest values of GRSP were recorded in the inoculated soils. Our findings suggest that the use of AM fungi and/or a PGPR for alleviating salinity stress in lettuce plants could be limited by their detrimental effect on soil structural stability.     

Rapid identification and discrimination among Egyptian genotypes of Rhizobium leguminosarum bv. viciae and Sinorhizobium meliloti nodulating faba bean (Vicia faba L.) by analysis of nodC, ARDRA, and rDNA sequence analysis

Twenty-eight Rhizobium strains were isolated from the root nodules of faba bean (Vicia faba L.) collected from 11 governorates in Egypt. A majority of these strains (57%) were identified as Rhizobium leguminosarum bv. viciae (Rlv) based on analysis of a nodC gene fragment amplified using specific primers for these faba bean symbionts. The strains were characterized using a polyphasic approach, including nodulation pattern, tolerance to environmental stresses, and genetic diversity based on amplified ribosomal DNA-restriction analysis (ARDRA) of both 16S and 23S rDNA. Analysis of tolerance to environmental stresses revealed that some of these strains can survive in the presence of 1% NaCl and a majority of them survived well at 37 °C. ARDRA indicated that the strains could be divided into six 16S rDNA genotypes and five 23S rDNA genotypes. Sequence analysis of 16S rDNA indicated that 57% were Rlv, two strains were Rhizobium etli, one strain was taxonomically related to Rhizobium rubi, and a group of strains were most closely related to Sinorhizobium meliloti. Results of these studies indicate that genetically diverse rhizobial strains are capable of forming N₂-fixing symbiotic associations with faba bean and PCR done using nodC primers allows for the rapid identification of V. faba symbionts.     

Root herbivory by Tipula paludosa larvae increases colonization of Agrostis capillaris by arbuscular mycorrhizal fungi

The objective of this study was to determine if root-feeding by insect larvae affects subsequent colonization of roots by arbuscular mycorrhizal fungi. We investigated grazing by larvae of Tipula paludosa on colonization of Agrostis capillaris by two species of fungi, Glomus mosseae and G. intraradices. Host plants were subjected to 7 days of grazing only, continuous grazing for 42 days, or no herbivory. Those plants with no herbivory had significantly lower levels of colonization by arbuscules and hyphae compared to plants which were grazed for 7 or 42 days. The effect only occurred in the upper parts of the root system, where larvae were active. We suggest that this effect was most likely mediated by a change in quantity and composition of root exudates.     

Stability of desiccated rhizosphere soil aggregates of mycorrhizal Juniperus oxycedrus grown in a desertified soil amended with a composted organic residue

Adequate soil structural stability favours the establishment and viability of a stable plant cover, protecting the soil against water erosion in desertified Mediterranean environments. We studied the effect of soil drying-rewetting, inoculation with a mixture of three exotic arbuscular mycorrhizal (AM) fungi (Glomus intraradices Schenck & Smith, Glomus deserticola (Trappe, Bloss. & Menge) and Glomus mosseae (Nicol & Gerd.) Gerd. & Trappe) and addition of a composted organic residue on aggregate stabilisation of the rhizosphere soil of Juniperus oxycedrus. The AM fungi and composted residue produced similar increases in plant growth, independently of the water conditions. Under well-watered conditions, the highest percentages of stable aggregates were recorded in the amended soil, followed by the soil inoculated with AM fungi. Excepting microbial biomass C, the soil drying increased labile C fractions (water soluble C, water soluble and total carbohydrates), whereas the rewetting decreased significantly such C fractions. Desiccation caused a significant increase in aggregate stability of the rhizosphere soil of all plants, particularly in the amended and inoculated plants. In all treatments, the aggregates formed after soil drying were unstable, since, in the rewetting, they disappear, reaching the initial levels before soil drying. Our results suggest that the aggregation mechanisms developed by rhizosphere microbial community of the amended and inoculated plants under water stress can be particularly relevant in desertified soils exposed to long desiccation periods.     

Using native plants to evaluate the effect of arbuscular mycorrhizal fungi on revegetation of iron tailings in grasslands

A pot experiment was conducted to evaluate native plant species associated with exogenous AMF for their suitability in the revegetation of iron mine tailings of Inner Mongolia grassland. Agropyron cristatum (L.) Gaertn. and Elymus dahuricus Turcz. associated with AMF, Glomus mosseae, or Glomus versiforme, were grown on iron mine tailings to assess the mycorrhizal effects on plant growth, mineral nutrition uptake, C:N:P stoichiometry, and heavy metals uptake. The symbiotic associations were successfully established between exogenous AMF and two native plants, and root colonization rates of G. versiforme were significantly (P?<?0.05) higher than those of G. mosseae. G. versiforme was more effective than G. mosseae in promoting plant growth by significantly (P?<?0.05) increasing the concentrations of N, P, and K and decreasing the ratios of C:N:P. The shoot and root dry weights of A. cristatum and E. dahuricus were increased by 51–103 %. The N, P, and K concentrations of shoots and roots of two plants were increased by 18–236 %. Inoculation with AMF also significantly (P?<?0.05) decreased concentrations of heavy metals in the shoots and increased those in the roots, indicating that AMF could confer some degree of heavy metal tolerance to plants. The results indicated that plant inoculation with G. versiforme was more suitable than inoculation with G. mosseae for the revegetation of iron mine tailings. The experiment provided evidence for the potential use of local plant species in combination with exogenous AMF for ecological restoration of metalliferous tailings in arid and semi-arid grassland.     

Pre-inoculation with arbuscular mycorrhizal fungi suppresses root knot nematode (<Emphasis Type="Italic">Meloidogyne incognita</Emphasis>) on cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis>)

A pot experiment was conducted to evaluate the influence of pre-inoculation of cucumber plants with each of the three arbuscular mycorrhizal (AM) fungi Glomus intraradices, Glomus mosseae, and Glomus versiforme on reproduction of the root knot nematode Meloidogyne incognita. All three AM fungi tested significantly reduced the root galling index, which is the percentage of total roots forming galls. Numbers of galls per root system were significantly reduced only in the G. intraradices + M. incognita treatment. The number of eggs per root system was significantly decreased by AM fungus inoculation, no significant difference among the three AM fungal isolates. AM inoculation substantially decreased the number of females, the number of eggs g⁻¹ root and of the number of eggs per egg mass. The number of egg masses g⁻¹ root was greatly reduced by inoculation with G. mosseae or G. versiforme. By considering plant growth, nutrient uptake, and the suppression of M. incognita together, G. mosseae and G. versiforme were more effective than G. intraradices.     

Glycosidation of apigenin results in a loss of its activity on different growth parameters of arbuscular mycorrhizal fungi from the genus Glomus and Gigaspora

The effect of different concentrations (0.5, 2 and 8 μM) of apigenin and its glycosidated form 5,7,4′-hydroxy flavone glycoside on arbuscular mycorrhizal (AM) fungal spore germination, hyphal growth, hyphal branching, the formation of entry points and root colonization of Gigaspora. rosea, Gi. margarita, Glomus mosseae and G. intraradices was tested. The lowest apigenin concentration (0.5 μM) nearly doubled hyphal branching, the formation of entry points and root colonization of all four tested fungi, whereas higher concentrations (2 and 8 μM) nearly doubled the hyphal growth of Gi. margarita, G. mosseae and G. intraradices. In none of the treatments with the apigenin-glycoside any effect on AM fungi could be observed. Our data show that apigenin exhibits an AM fungal genus and even species activity and we provide strong evidence that glycosidation results in a loss of its activity towards AM fungi.     

AM fungi inoculation and addition of microbially‐treated dry olive cake‐enhanced afforestation of a desertified Mediterranean site

A field experiment was carried out to compare the effectiveness of inoculation with three arbuscular mycorrhizal (AM) fungi, namely Glomus intraradices Schenck & Smith, Glomus deserticola (Trappe, Bloss. & Menge) and Glomus mosseae (Nicol & Gerd.) Gerd. & Trappe, and the addition of Aspergillus niger‐treated dry olive cake (DOC) in the presence of rock phosphate, in increasing root nitrate reductase (NR) and acid phosphatase activities, mycorrhizal colonization, plant growth and nutrient uptake in Dorycnium pentaphyllum L. seedlings afforested in a semiarid degraded soil. Three months after planting, both the addition of fermented DOC and the mycorrhizal inoculation treatments had increased root NR activity significantly, particularly the inoculation with G. deserticola (by 75 per cent with respect to non‐inoculated plants), but they had no effect on root acid phosphatase. Mycorrhizal inoculation treatments with G. deserticola or G. mosseae on their own were even more effective than the addition of fermented DOC alone in improving the growth and (NPK) foliar nutrients of D. pentaphyllum plants. The combined treatment involving the application of microbially‐treated agrowastes and mycorrhizal inoculation with AM fungi, particularly with G. mosseae, can be proposed as a successful revegetation strategy for D. pentaphyllum in P‐deficient soils under semiarid Mediterranean conditions. Copyright © 2004 John Wiley & Sons, Ltd.     

The effect of acidity on the distribution and symbiotic efficiency of rhizobia in Lithuanian soils

The distribution and symbiotic efficiency of nodule bacteria Rhizobium leguminosarum_bv. trifolii F., Sinorhizobium meliloti D., Rhizobium galegae L., and Rhizobium leguminosarum bv. viciae F. in Lithuanian soils as dependent on the soil acidity were studied in the long-term field, pot, and laboratory experiments. The critical and optimal pH values controlling the distribution of rhizobia and the symbiotic nitrogen fixation were determined for every bacterial species. The relationship was found between the soil pH and the nitrogen-fixing capacity of rhizobia. A positive effect of liming of acid soils in combination with inoculation of legumes on the efficiency of symbiotic nitrogen fixation was demonstrated.     

Establishment of Retama sphaerocarpa L. seedlings on a degraded semiarid soil as influenced by mycorrhizal inoculation and sewage‐sludge amendment

A field experiment was carried out to evaluate the effectiveness of mycorrhizal inoculation with three arbuscular mycorrhizal (AM) fungi (Glomus intraradices Schenck & Smith, Glomus deserticola (Trappe, Bloss. & Menge), and Glomus mosseae (Nicol & Gerd.) Gerd. & Trappe) and the addition of composted sewage sludge (SS) with respect to the establishment of Retama sphaerocarpa L. seedlings, in a semiarid Mediterranean area. Associated changes in soil chemical (nutrient content and labile carbon fractions), biochemical (enzyme activities), and physical (aggregate stability) parameters were observed. Six months after planting, both the addition of composted SS and the mycorrhizal‐inoculation treatments had increased total N content, available‐P content, and aggregate stability of the soil. Values of water‐soluble C and water‐soluble carbohydrates were increased only in the mycorrhizal‐inoculation treatments. Rhizosphere soil from the mycorrhizal‐inoculation treatments had significantly higher enzyme activities (dehydrogenase, protease‐BAA, acid phosphatase, and β‐glucosidase) than the control soil. In the short‐term, mycorrhizal inoculation with AM fungi was the most effective treatment for enhancement of shoot biomass, particularly with G. mosseae (about 146% higher with respect to control plants). The addition of the composted SS alone was sufficient to restore soil structural stability but was not effective with respect to improving the performance of R. sphaerocarpa plants.     

The bioprotective effect of AM root colonization against the soil-borne fungal pathogen Gaeumannomyces graminis var. tritici in barley depends on the barley variety

The systemic effect of root colonization by the arbuscular mycorrhizal fungus (AMF) Glomus mosseae on the susceptibility of old and modern barley varieties to the soil-borne fungal pathogen Gaeumannomyces graminis var. tritici (Ggt) was studied in a split-root system. Plants were precolonized on one side of the split-root system with the AMF and thereafter the other side of the split-root system was inoculated with the pathogen. At the end of the experiment the level of bioprotection was estimated by quantifying lesioned roots and the determination of the root fresh weight. AM root colonization provided protection in some of the barley genotypes tested, but not in others. This protective effect seemed to vary in the oldest and the most modern barley variety tested.     

Differences between strains of rhizobium trifolii in ability to colonize soil and plant roots in the absence of their specific host plants

Evidence is presented of strain differences within Rhizobium trifolii in ability to colonize soil surrounding common pasture species, both legume and non-legume, in problem sandy soils in Western Australia. R. lupini and R. trifolii are shown to spread better than R. meliloti in these soils.