Zinc influences nodulation,disease severity,leaf drop and herbage yield of alfalfa cultivars

Zinc deficiency is an important limiting factor in sustainable crop production and is a factor often overlooked in determining the benefits and overall success of alfalfa pastures in rotations. A field experiment was conducted to investigate the effects of zinc and alfalfa cultivars on nodulation, herbage yield, leaf drop and disease severity (Phytophthora root rot disease and common leaf spot disease) in alfalfa (Medicago sativa L.). Ten cultivars of alfalfa (Hunter River, Hunterfield, Sceptre Aurora, Genesis, Aquarius, Venus (Y8622), PL69, P5929 and PL34HQ) were tested at two levels of zinc (+Zn: 4 kg ha^–1, -Zn: no zinc added) on a Zn-deficient soil (DTPA zinc of top 0–15 cm soil was 0.4 mg kg^–1 soil, while 15–30 cm subsoil Zn was 0.1 mg kg^–1 soil) under field conditions. Zinc application significantly increased number and dry weight of nodules, herbage yield and leaf to stem ratio of alfalfa plants. There was a significant reduction in leaf drop, and occurrence of Phytophthora root rot (caused by fungus Phytophthora megasperma f. sp. medicaginis) and common leaf spot (caused by fungus Pseudopeziza medicaginis) diseases with Zn application. Alfalfa cultivars had a differential response to low Zn. Hunter River and Hunterfield were the most affected by omitting zinc application, while Sceptre, PL34HQ and Aquarius were comparatively less affected. The present study suggest that Zn nutrition effects nodulation, leaf drop, disease occurrence and production potential of alfalfa. The alfalfa cultivars have differential ability to low Zn supply. Growing of Zn-efficient cultivars and adequate Zn nutrition may also improve the N₂-fixation by alfalfa on low-Zn soils.     

Interaction between alfalfa cultivars and Rhizobium strains for nitrogen Fixation

Summary Two experiments were conducted in the greenhouse to study the interaction between alfalfa cultivars (Medicago sativa L. and M. falcata L.) and strains of Rhizobium meliloti Dang. for acetylene reduction rate, plant height and dry weights of shoot, root and whole plant. Fifteen alfalfa cultivars were inoculated with 10 strains of Rhizobium in Experiment I. Variance component analysis revealed that more than 30% of the total variance was due to alfalfa cultivars for acetylene reduction rate and 26% was accounted for by Rhizobium strains. More than 36% of the total variation was attributed to the interaction between alfalfa cultivars and Rhizobium strains for this character. Twenty-five host cultivars and 11 Rhizobium strains were included in Experiment II. The results also showed that the interaction of alfalfa cultivars and Rhizobium strains contributed the largest portion of the total variation for dry weights of shoot, root and whole plant and acetylene reduction rate. The results clearly demonstrated that the non-additive effects were the major component of variation for these characters associated with nitrogen fixation in alfalfa. Therefore, an effective way of improving nitrogen fixation in alfalfa is to select for a favourable combination of specific Rhizobium strains and alfalfa cultivars.     

Associative effect of Rhizobium and phosphate-solubilizing bacteria on the yield and nutrient uptake of chickpea

Combined inoculation of Rhizobium and ‘Phosphate-solubilizing’Pseudomonas striata orBacillus polymyxa with and without added chemical fertilizer on chickpea yield and nutrient content was studied under greenhouse conditions. While the single inoculation of Rhizobium increased the nodulation and nitrogenase activity, the ‘phosphate-solubilizers’ increased the available phosphorus content of the soil. Combined inoculation of Rhizobium andP. striata orB. polymyxa increased the above parameters and also the dry matter content, the grain yield and nitrogen and phosphorus uptake significantly over the uninoculated control. The inoculation effects were more pronounced in the presence of added fertilizers. The possibilities of saving half the dose of N and replacing superphosphate with rockphosphate and inoculation with ‘phosphate-solubilizers’ are discussed.     

Rhizobium meliloti inoculation of alfalfa selected for tolerance to acid,aluminum-rich soils

Alfalfa (Medicago sativa L.) growth and nodulation in acid soil is reduced because the plant and its bacterial symbiontRhizobium meliloti cannot tolerate acid, aluminum-rich soil. A study was conducted to determine if a relatively acid-tolerant alfalfa germplasm combined with a relatively acid-tolerantR. meliloti strain could overcome these limitations. In a light room study, an acid-tolerant alfalfa germplasm inoculated with a more acid-tolerantR. meliloti strain produced greater top growth, nodule number and weight, and acetylene reduction values in an unlimed soil (pH 4.6) than the same germplasm inoculated with a relatively acid-sensitiveR. meliloti strain or an acid-sensitive germplasm inoculated with either a relatively acid-tolerant or acid-sensitiveR. meliloti strain.     

Influence of drought on competition between selected Rhizobium meliloti strains and naturalized soil rhizobia in alfalfa

Drought is an important environmental factor that can affect rhizobial competition and N₂ fixation. Three alfalfa (Medicago sativa L. and M. falcata L.) accessions were grown in pots containing soil from an irrigated (Soil 1) and a dryland (Soil 2) alfalfa field in northern Utah, USA. Mutants of three strains of Rhizobium meliloti Dang. from Pakistan (UL 136, UL 210, and UL 222) and a commercial rhizobial strain 102F51a were developed with various levels of resistance to streptomycin. Seeds inoculated with these individual streptomycin-resistant mutants were sown in the two soils containing naturalized rhizobial populations. Soils in the pots were maintained at −0.03, −0.5, and −1.0 MPa. After 10 weeks, plants were harvested and nodule isolates were cultured on agar medium with and without streptomycin to determine nodule occupancy (proportion of the nodules occupied by introduced rhizobial strains). Number of nodules, nodule occupancy, total plant dry weight, and shoot N were higher for Soil 1 than Soil 2. Number of nodules, plant dry weight, and shoot N decreased as drought increased from −0.03 to −1.0 MPa in the three alfalfa accessions. Rhizobial strains UL 136 and UL 222 were competitive with naturalized alfalfa rhizobia and were effective at symbiotic N₂ fixation under drought. These results suggest that nodulation, growth, and N₂ fixation in alfalfa can be improved by inoculation with competitive and drought-tolerant rhizobia and may be one economically feasible way to increase alfalfa production in water-limited environments. Joint contribution from USDA-ARS and the Utah Agric. Exp. Sta., Utah State Univ., Logan, UT 84322-4810, USA. Journal Paper No. 4931. Joint contribution from USDA-ARS and the Utah Agric. Exp. Sta., Utah State Univ., Logan, UT 84322-4810, USA. Journal Paper No. 4931.     

Root exuded nod-gene inducing signals limit the nodulation capacity of different alfalfa varieties with Rhizobium meliloti

Summary Different nodulation capacities were found among nine different varieties of alfalfa, cultivated in the Central region of Mexico, by Rhizobium meliloti 2011. A correlation between nodulation capacity and foliar dry weight was observed, which points to a genotype dependance on these parameters. A correlation between the nodulation capacity and the R. meliloti nod-gene inducing activity of the root exudates from the different varieties, as measured by -galactosidase induction in a test system consisting of a R. meliloti nodC-lacZ strain incubated with each root exudate, was established. When the root exudate from the best nodulating variety was added to the four poorest nodulating varieties, an increase in nodule formation was observed. We conclude that root exuded nod-gene inducing signals are a symbiotically-limiting component in natural populations of the poorest nodulating varieties of alfalfa.     

Net assimilation rate and relative nitrogen assimilation rate in relation to the dry matter production of alfalfa cultivars

Summary Twelve alfalfa cultivars inoculated with an indigenous strain (RM9) ofRhizobium meliloti, were compared for their seedling morphological characters, and growth characters, including net assimilation rate (NAR), relative growth rate (RGR), leaf area ratio (LAR) and relative nitrogen assimilation rate (R_N). Highly significant differences were obtained between cultivars for most characters.Simple correlation showed that NAR influenced RGR (r=0.91) more than leaf area ratio (LAR) (r=–0.44), and that most characters measured were highly correlated with seedling dry weight. Factor analysis showed that NAR, RGR and R_N contributed 25% of the total variation in the dependence structure. The grouping indicated that the higher the NAR and R_N the greater was the RGR. Path-coefficient analysis showed that NAR had more important direct and indirect effects than R_N in dry matter accumulation. The relationship implied that selection for plants with high NAR, or high efficiency in converting light energy to dry matter production could contribute greater N₂ fixation in alfalfa.     

Nitrogen-fixing characteristics of alfalfa cultivars nodulated by representatives of an indigenousRhizobium meliloti serogroup

Studies were conducted to evaluate whether field-grown cultivars of alfalfa (Medicago sativa L.) nodulate differentially with members of a soil population ofRhizobium meliloti, and to determine the influence of the dominant nodule occupants on N₂-dependent growth of the same cultivars under greenhouse conditions. Nodules were sampled from four replicate plots of Vernal, Anchor, and Saranac alfalfa, and the isolates analysed serologically. Results from agglutination tests identified serogroup 31 as a dominant nodule occupant. A significant cultivar effect was observed, with a greater and more consistent occupancy rate by serogroup 31 across the replicates on Vernal (60%) compared to Anchor (24%) or Saranac (36%). The symbiotic effectiveness of the parent isolate of serogroup 31 was evaluated on each cultivar over four successive harvests in a greenhouse study. Significant cultivar x N source interactions for herbage dry weight resulted following the second harvest. Of the three cultivars, only inoculated Vernal responded with an increase in shoot dry weight and N₂ assimilated relative to N supplemented plants between harvests two and three. In separate greenhouse experiments, field isolates of serogroup 31 from nodules on Vernal produced homogeneous, effective responses both on Vernal and Anchor. In contrast, serogroup 31 field isolates from Anchor nodules were highly heterogeneous in effectiveness on the parent host, with poorly effective isolates being substantially more effective on Vernal. The data indicate that attention should be given to the potential impact of the indigenousR. meliloti population upon cultivar ranking at specific field locations, and also to strain-cultivar idiosyncracies when carrying out physiological sutidies of regrowth characteristics.Technical Paper No. 8716 of the Oregon State University Agricultural Experiment Station.     

Identification and characterization of common bean (Phaseolus vulgaris L.) lines well nodulated in the presence of high nitrate

Common bean,Phaseolus vulgaris L., is known to be ‘inefficient’ in nodulation and N₂ fixation although it responds to applied nitrogen. An experiment was conducted to identify and to characterize bean cultivars nodulating in the presence of a high level of nitrogen. Sixteen cultivars and a check for inefficient nodulation, OAC Seaforth, were inoculated and grown for 40 days in replicated pots supplied with zero, 3.5 and 10.5 mM combined nitrogen as NO ₃ ⁻ and NH ₄ ⁺ . Seven traits relating to nodulation and N₂ fixation were all significantly affected by N level (N), cultivar (Cv) and N × Cv interactions (except for root dry weight), indicating that cultivars responded differently to the N treatments. Total dry weight (W) and shoot to root ratio (S/R) increased with the increased N levels. Nodule dry weight (Wn), visual nodulation score (Nv) and nodulation index (Nx) decreased as the N increased. Percent N and N content per plant increased with the increased N level. Plant weight (W) was positively correlated with Wn, Nv and N content and negatively correlated with %N. Nodulation score was positively associated with Wn and plant N content. Genotypes superior in nodulation and N₂ fixation in the presence of N were identified. Cultivars Italian Barlotti, California Light Red Kidney, Kentucky Wonder A and Pueblo 152 were selected for further testing and use in improving the nitrate tolerant nodulating characteristic of bean.     

Rhizobium meliloti distribution in the soil following alfalfa inoculation

Summary Alfalfa seeds, inoculated with an antibiotic-resistantRhizobium meliloti strain, were planted in three replicated field plots at Clayton, N.C. Core samples were taken three times in the next year at 0, 10, and 20 cm from the edge of each plot. Soil subsamples were taken from within each core sample at 0, 6, 12, and 18 cm depths. The numbers of the inoculum Rhizobium strain in each soil subsample were determined by inoculation of alfalfa plants with diluted soil samples. In general the distribution of rhizobia showed some movement outward and downward in the soil. Lower counts were obtained at the surface during summer. The Rhizobium persistence pattern in the soil differed in the three plots which is consistent with the variability in Rhizobium numbers often observed in established alfalfa stands. Cooperative investigation of the United States Department of Agriculture, Science and Education Administration, Agricultural Research and the North Carolina Agricultural Research Service, Raleigh, North Carolina. Paper No. 6818 of the Journal Series of the North Carolina Agricultural Research Service at Raleigh.     

Influence of soil pH on the soybean-endomycorrhiza symbiosis

Summary Soybean (Glycine max {L.} Merr.) cultivars were inoculated withGigaspora gigantea andGlomus mosseae to determine mycorrhizal: cultivar relationships as affected by soil pH. The specific cultivarfungal response was dependent on soil pH. Overall cultivar responses in unlimed soil (pH 5.1) were greater forG. gigantea thanG. mosseae. The Bossier —G. gigantea combination was particularly responsive in unlimed soil and showed an increase of 10% in shoot length, 35% in shoot dry weight. 75% in root dry weight, and 397% in nodule dry weight over uninoculated controls. Little cultivar response was observed withG. mosseae inoculation in unlimed soil. In limed soil (pH 6.2), the larger responses were obtained withG. mosseae inoculated plants, although inoculation with eitherG. mosseae orG. gigantea appeared effective. In general, nodulation was greater on mycorrhizal roots than on control roots.     

Back-cross reciprocal monosomic analysis of Fusarium head blight resistance in wheat (Triticum aestivum L.)

 Fusarium head blight (FHB or scab) caused by Fusarium spp. is a widespread disease of cereals causing yield and quality losses and contaminating cereal products with mycotoxins. The breeding of resistant varieties is the method of choice for controlling the disease. Unfortunately, the genetic basis of scab resistance is still poorly understood. We present the results of a back-cross reciprocal monosomic analysis of FHB resistance using the highly resistant Hungarian winter wheat line ‘U-136.1’ and the highly susceptible cultivar ‘Hobbit-sib’. Resistance testing was performed in a field trial artificially inoculated with a Fusarium culmorum conidial suspension. Five hemizygous families containing ‘U-136.1’ chromosomes 6B, 5A, 6D, 1B, and 4B had a visually reduced spread of infection compared to lines having the ‘Hobbit-sib’ chromosome. Chromosome 2B from ‘U-136.1’ had an increased spread of infection. The critical chromosomes controlling seed weight were 6D, 3B, 5A, and 6B while those controlling deoxynivalenol (DON) content were homoeologous groups 2 and 6, although the latter effects were not significant due to a high coefficient of variation. Results from this and other studies show that chromosomes 6D, 6B, 5A, 4D, and 7A have frequently been associated with scab resistance in a number of wheat cultivars. Research groups now attempting to map scab resistance in wheat using markers should pay special attention to the above-mentioned chromosomes. Received: 31 March 1998 / Accepted: 14 July 1998     

Nitrogen release from roots of alfalfa and soybean grown in sand culture

An enclosed root chamber containing sterile sand medium was used to study net nitrogen (N) release from actively growing root systems of ‘Saranac’ alfalfa (Medicago sativa L.) and ‘Fiskeby V’ soybean (Glycine max L. Merr.). Plants were inoculated with a rhizobial strain appropriate to each host, irrigated with N-free nutrient solution, and grown either to 85 or to 173 d after germination (alfalfa) or to physiological maturity (soybean). Alfalfa released 4.5% of symbiotically-fixed plant N into the root zone over its growth period; soybean released 10.4% of plant N. Root zone leachates were analyzed for total N and for amino acid and ammonium content. Significant ammonium-N release occurred from the alfalfa but not the soybean root system; little amino-N was released by root systems of either species. Shoot harvest and water deficit caused increased release of N from alfalfa roots. The results provide evidence that alfalfa and soybean released significant proportions of their N into the root zone, and indicate that while substantial ammonium-N was released from alfalfa roots, passive leakage of amino-N was not a primary mechanism for N release from root systems of either species. Cooperative investigation of USDA-ARS and the Minnesota Agric. Exp. Stn. (Scientific J. Series No. 16048). This research was supported in part by a Graduate School Fellowship to LSB from the Univ. of Minnesota.     

The contributions of plant and bacteria genotypes in the growth and nitrogen accumulation of inoculated alfalfa

The symbiotic efficiency of each of 30 alfalfa (16 Medicago sativa and 14 M. varia) cultivars inoculated with 7 Rhizobium meliloti strains was studied in three field experiments. Two-factor analysis of variance of the obtained date demonstrated that the green mass yield and nitrogen accumulation depend on genotypes of both partners. The total contribution of plant and bacterial genotypes to the variation of green mass yield increased from 0–17% in the first year of alfalfa growth to 40–78% in the third year. The effect of the genotypic variability of the symbiotic partners was higher for N accumulation than for the green mass. There was a negative correlation between plant mass and N accumulation in the uninoculated plants with the relative (%) deviations of these parameters in the inoculated plants. In the experiments conducted in the Tashkent region the efficiency of the “alfalfa-R. meliloti” symbiosis was higher than in the experiment conducted in the Tumen region.     

Field resistance of spring barley cultivars to powdery mildew in Lithuania

During vegetative period 2004–2005 powdery mildew (Erysiphe graminis DC. f. sp. hordei Em. Marchal) field resistance of spring barley cultivars was investigated at the Lithuanian Institute of Agriculture. The spring barley genotypes tested were Lithuania-registered cultivars, cultivars from genetic resources collection, and the new cultivars used for initial breeding. In total, 23 resistance genes were present in the 84 cultivars studied. Among mono-genes only mlo and 1-B-53 showed very high resistance. Slight powdery mildew necroses (up to 3 scores) formed on cultivars possessing these genes. The maximal powdery mildew (PM) severity reached a score of 8.5 and the area under disease progress curve (AUDPC) a value of 1216.8. The cultivars ‘Primus’, ‘Astoria’, ‘Power’, ‘Harrington’ and ‘Scarlett’ were the most resistant among the non mlo cultivars. Severity of PM on ‘Primus’ reached a score of 3.5 (3.0 of PM necrosis) in average, the other cultivars were diseased from 4.5 (3.0) to 5.0 (2.0). The AUDPC values for these cultivars except ‘Scarlett’ were the lowest (85.0–145.3) among the other cultivars. The highest contrast in development of the other leaf diseases was between highly resistant and susceptible to PM cultivar groups. The fast development of PM depressed development of the other diseases 4.7 times.     

Response of Leucaena/Rhizobium symbiosis to mineral nutrients in Southwestern Nigeria

Pot and field experiments carried out at the International Institute of Tropical Agriculture (IITA) and at Fashola, Southwestern Nigeria, examined the effect of inoculation and N, P and micronutrients on nodulation and growth ofLeucaena leucocephala (Lam.) de Wit. In pot studies all parameters measured, except the percentage of nitrogen in shoots, were improved by inoculation, nitrogen and phosphorus. Micronutrients increased only nitrogen and allantoin contents. Interactions between inoculation and P, N and micronutrients on nodulation and growth of leucaena were observed. The effect of inoculation and fertilization with phosphorus or micronutrients was further investigated in field experiments. Establishment of uninoculated and unfertilized leucaena was poor at both locations due to low soil fertility and the presence of only a few native leucaena rhizobia. At one site, only inoculated plants were nodulated, while at the other, all plants produced nodules. Shoot dry weight, total nitrogen and phosphorus of inoculated plants were statistically equal to nitrogen-fertilized plants. Uninoculated plants were stunted. Generally, micronutrients did not influence nodulation, total nitrogen or growth of leucaena. They had only a positive effect on nitrogenase activity. Phosphorus increased total nitrogen and phosphorus uptake and plant growth. A 75% increase in shoot dry weight was obtained when 80 kg P ha^–1 was applied to inoculated leucaena with Rhizobium strain IRc 1045. Inoculated plants contained more allantoins than uninoculated ones but no significant correlation was found between these compounds and other parameters of N fixation.     

RFLP variation in diploid and tetraploid alfalfa

Summary Alfalfa (Medicago sativa L.) is a major forage crop throughout the world. Although alfalfa has many desirable traits, continued breeding is required to incorporate pest resistances and other traits. We conducted this study to determine the amount of restriction fragment length polymorphism (RFLP) variability present within and between diploid and tetraploid alfalfa populations, and whether or not this variability is sufficient for construction of an RFLP map. Diploid plants from M. sativa ssp. falcata, ssp. coerulea, and ssp. sativa and tetraploid spp. sativa cultivars Apollo, Florida 77, and Spredor 2 were included. A total of 19 cDNA clones was probed onto genomic Southern blots containing DNA digested by EcoRI, HindIII, or BamHI. Phylogenetic trees were produced, based on parsimony analysis of shared restriction fragments. Evidence for extensive gene duplication was found; most probes detected complex patterns of restriction fragments. Large amounts of variation are present within all diploid subspecies. M. sativa ssp. falcata plants formed clusters distinct from ssp. sativa or ssp. coerulea plants, which were not distinctly clustered. Some M. sativa ssp. falcata plants were more similar to the other groups than to other plants within ssp. falcata. Variation among tetraploid cultivars showed that Florida 77 and Apollo had more similarities than either showed with Spredor 2. All three cultivars showed large within-population variation, with Apollo being the most diverse and Spredor 2 the least. Based on these results, development of an RFLP map at the diploid level appears possible. Also, differentiation of cultivars, particularly ones of divergent origin, seems possible based on RFLP patterns.     

Response of strawberry to inoculation with arbuscular mycorrhizal fungi under very high soil phosphorus conditions

A field study was done to assess the potential benefit of arbuscular mycorrhizal (AM) inoculation of elite strawberry plants on plant multiplication, under typical strawberry nursery conditions and, in particular, high soil P fertility (Mehlich-3 extractible P=498 mg kg⁻¹). Commercially in vitro propagated elite plants of five cultivars (‘Chambly,’ ‘Glooscap,’ ‘Joliette,’ ‘Kent,’ and ‘Sweet Charlie’) were transplanted in noninoculated growth substrate or in substrate inoculated with Glomus intraradices or with a mixture of species (G. intraradices, Glomus mosseae, and Glomus etunicatum) at the acclimation stage and were grown for 6 weeks before transplantation in the field. We found that AM fungi can impact on plant productivity in a soil classified as excessively rich in P. Inoculated mother plants produced about 25% fewer daughter plants than the control in Chambly (P=0.03), and Glooscap produced about 50% more (P=0.008) daughter plants when inoculated with G. intraradices, while the productivity of other cultivars was not significantly decreased. Daughter plant shoot mass was not affected by treatments, but their roots had lower, higher, or similar mass, depending on the cultivar–inoculum combination. Root mass was unrelated to plant number. The average level of AM colonization of daughter plants produced by noninoculated mother plants did not exceed 2%, whereas plants produced from inoculated mothers had over 10% of their root length colonized 7 weeks after transplantation of mother plants and ∼6% after 14 weeks (harvest), suggesting that the AM fungi brought into the field by inoculated mother plants had established and spread up to the daughter plants. The host or nonhost nature of the crop species preceding strawberry plant production (barley or buckwheat) had no effect on soil mycorrhizal potential, on mother plant productivity, or on daughter plant mycorrhizal development. Thus, in soil excessively rich in P, inoculation may be the only option for management of the symbiosis.     

Evaluation of the symbiotic nitrogen-fixing potential of soils by direct microbiological means

A method for estimating the nitrogen-fixing capacity of a population of rhizobia resident in soil is presented. legume test plants, growing under microbiologically-controlled conditions in test tubes packed with a vermiculite substrate moistened with a nitrogen-free plant nutrient solution, are inoculated directly with a suspension of the soil under examination. Rhizobia in the soil nodulate the test plants, and the amount of foliage dry matter produced in the 28 days after inoculation is regarded as an index of their effectiveness. An inoculum of at least 30, and preferably 100, rhizobia is needed to ensure that nitrogen fixation is not masked by delayed nodulation. The new method is tentatively described as the ‘whole-soil inoculation’ technique. Appraisals were made withTrifolium subterraneum L. andRhizobium trifolii and withMedicago sativa L. andR. meliloti. Soil-borne pathogens did not interfere with plant growth. The whole-soil inoculation technique was less tedious and time-consuming than an alternative method which involved extracting representative isolates from the soil and testing their effectiveness individually, and appeared to give more realistic values for the nitrogen-fixing capacity of the soil as a whole. Used in association with a field experiment, the whole-soil inoculation technique confirmed microbiologically that there had been an agronomic response to surface application of inoculant to poorly-nodulatedT. subterraneum pasture. It is submitted that this technique for determining the effectiveness of rhizobia in soil, combined with a plant-infection method for counting rhizobia, can be a reliable guide to the need for inoculation in the field.     

Co-inoculation with antibiotic-producing bacteria to increase colonization and nodulation by rhizobia

A study was conducted to determine whether colonization of legume roots and nodulation byRhizobium meliloti andBradyrhizobium japonicum could be enhanced by using inocula containing microorganisms that produce antibiotics suppressing soil or rhizosphere inhabitants but not the root-nodule bacteria. An antibiotic-producing strain of Pseudomonas and one of Bacillus were isolated, and mutants ofR. meliloti andB. japonicum sp. resistant to the antibiotics were used. The colonization of the alfalfa rhizosphere and nodulation byR. meliloti were enhanced by inoculation of soil withPseudomonas sp. in soil initially containing 2.7×10⁵ R. meliloti per g. The colonization of soybean roots byB. japonicum was enhanced by inoculating soil with three cell densities ofBacillus sp., and nodulation was stimulated byBacillus sp. added at two cell densities. In some tests, the dry weights of soybeans and seed yield increased as a result of these treatments, and co-inoculation with Bacillus also increased pod formation. Inoculation of seeds withBacillus sp. and the root-nodule bacterium enhanced nodulation of soybeans and alfalfa, but colonization byB. japonicum andR. meliloti was stimulated only during the early period of plant growth. Studies were also conducted withStreptomyces griseus and isolates ofR. meliloti andB. japonicum resistant to products of the actinomycete. Nodulation of alfalfa byR. meliloti was little or not affected by the actinomycete alone; however, both nodulation and colonization were enhanced if the soil was initially amended with chitin andS. griseus was also added. Chitin itself did not affectR. meliloti. Treatments of seeds with chitin orS. griseus alone did not enhance colonization of alfalfa roots byR. meliloti or soybean roots byB. japonicum, but the early colonization of the roots by both bacterial species was promoted if the seeds received both chitin andS. griseus; this treatment also increased nodulation and dry weights of alfalfa and soybeans and the N content of alfalfa. It is suggested that co-inoculation of legumes with antibiotic-producing microorganisms and root-nodule bacteria resistant to those antibiotics is a promising means of promoting nodulation and possibly nitrogen fixation.