Evolutionary history shapes patterns of mutualistic benefit in Acacia–rhizobial interactions

The ecological and evolutionary factors that drive the emergence and maintenance of variation in mutualistic benefit (i.e., the benefits provided by one partner to another) in mutualistic symbioses are not well understood. In this study, we evaluated the role that host and symbiont phylogeny might play in determining patterns of mutualistic benefit for interactions among nine species of Acacia and 31 strains of nitrogen‐fixing rhizobial bacteria. Using phylogenetic comparative methods we compared patterns of variation in mutualistic benefit (host response to inoculation) to rhizobial phylogenies constructed from housekeeping and symbiosis genes; and a multigene host phylogeny. We found widespread genotype‐by‐genotype variation in patterns of plant growth. A relatively large component of this variation (21–28%) was strongly influenced by the interacting evolutionary histories of both partners, such that phylogenetically similar host species had similar growth responses when inoculated with phylogenetically similar rhizobia. We also found a relatively large nonphylogenetic effect for the average mutualistic benefit provided by rhizobia to plants, such that phylogenetic relatedness did not predict the overall benefit provided by rhizobia across all hosts. We conclude that phylogenetic relatedness should frequently predict patterns of mutualistic benefit in acacia‐rhizobial mutualistic interactions; but that some mutualistic traits also evolve independently of the phylogenies.     

滨海盐渍土野大豆根瘤菌分离筛选及其在大豆中的应用

【目的】了解盐渍土野大豆根瘤菌的多样性，筛选具有耐盐促生作用的菌株，为栽培大豆耐盐菌剂的开发提供菌种资源。【方法】采用传统培养方法从滨海盐渍土野大豆中分离根瘤菌，评价菌株的促生特性，并验证其对野大豆和栽培大豆的促生效果。【结果】从野大豆根和根瘤样品中分离出87株根瘤菌，主要为中华根瘤菌属(Sinorhizobium)、根瘤菌属(Rhizobium)和慢生根瘤菌属(Bradyrhizobium)。测定了24株代表性菌株的促生特性，发现有16株根瘤菌具有产吲哚-3-乙酸(indole-3-acetic acid, IAA)能力，6株能够产生1-氨基环丙烷-1-羧酸(1-amino-cyclopropane-1- carboxylic, ACC)脱氨酶，16株具有溶磷活性，6株能够产生铁载体。根据以上促生特性，选择了11株优良根瘤菌进行野大豆促生和结瘤能力评价，发现美洲中华根瘤菌(Sinorhizobium americanus) DL3的性能优于其他菌株。最后，通过盆栽试验检测了菌株DL3对野大豆和栽培大豆耐盐能力的影响，发现菌株DL3在盐胁迫下能促进野大豆和大豆的生长，同时，降低了叶片脯氨酸水平，缓解了植物的盐胁迫程度。【结论】菌株DL3在提高植物耐盐性方面具有一定的作用，对实现大豆的盐碱地种植具有重要的理论意义和实践价值。     

Growth Tolerance of Rhizobia Isolated from Sand Dune Legumes of the Southwest Coast of India

This paper describes the properties of rhizobia from extreme soil environments which are characterized by high temperatures, salt concentrations and also rather extreme pH values due to the contamination by spray water from the sea. Coastal sand dunes are such extreme habitats which support a variety of microorganisms. To explore stress‐tolerant rhizobia, ten rhizobial strains were isolated from five wild legumes from two dune systems of the southwest coast of India. They were tested for growth performance or tolerance at a wide range of temperatures (30–55 °C), salinity (0.1–4.5 % w/v) and initial pH values (3.5–11). Growth of five isolates was highest between 30–40 °C, while four isolates showed considerable growth up to 2.5 % salinity (at 35 °C). All isolates demonstrated elevated growth at an initial pH of between 5–6 (at 35 °C and 2 % salinity), while five isolates had additional growth peaks at an initial pH of between pH 7.5–9 indicating alkaline tolerance and were suitable for efficient phosphate solubilization. The stress tolerance traits of these rhizobia are of potential value for strain improvement in agriculture or the bioremediation of soils at elevated temperatures, salinity and extreme pH values, and thus are of high biotechnological importance.     

Effects of salt stress and rhizobial inoculation on growth and nitrogen fixation of three peanut cultivars

Increasing soil salinity represents a major constraint for agriculture in arid and semi‐arid lands, where mineral nitrogen (N) deficiency is also a frequent characteristic of soils. Biological N fixation by legumes may constitute a sustainable alternative to chemical fertilisation in salinity‐affected areas, provided that adapted cultivars and inoculants are available. Here, the performance of three peanut cultivars nodulated with two different rhizobial strains that differ in their salt tolerance was evaluated under moderately saline water irrigation and compared with that of N‐fertilised plants. Shoot weight was used as an indicator of yield. Under non‐saline conditions, higher yields were obtained using N fertilisation rather than inoculation for all the varieties tested. However, under salt stress, the yield of inoculated plants became comparable to that of N‐fertilised plants, with minor differences depending on the peanut cultivar and rhizobial strain. Our results indicate that N fixation might represent an economical, competitive and environmentally friendly choice with respect to mineral N fertilisation for peanut cultivation under moderate saline conditions.     

Phenotypic and molecular characterization of indigenous rhizobia nodulating chickpea in India

In a combined approach of phenotypic and genotypic characterization, 28 indigenous rhizobial isolates obtained from different chickpea growing regions in peninsular and northern India were analyzed for diversity. The field isolates were compared to two reference strains TAL620 and UPM-Ca142 representing M. ciceri and M. mediterraneum respectively. Phenotypic markers such as resistance to antibiotics, tolerance to salinity, temperature, pH, phosphate solubilization ability, growth rate and also symbiotic efficiency showed considerable diversity among rhizobial isolates. Their phenotypic patterns showed adaptations of rhizobial isolates to abiotic stresses such as heat and salinity. Two salt tolerant strains (1.5% NaCl by T1 and T4) with relatively high symbiotic efficiency and two P-solubilising strains (66.7 and 71 microg/ml by T2 and T5) were identified as potential bioinoculants. Molecular profiling by 16S ribosomal DNA Restriction Fragment Length Polymorphism (RFLP) revealed three clusters at 67% similarity level. Further, the isolates were differentiated at intraspecific level by 16S rRNA gene phylogeny. Results assigned all the chickpea rhizobial field isolates to belong to three different species of Mesorhizobium genus. 46% of the isolates grouped with Mesorhizobium loti and the rest were identified as M. ciceri and M. mediterraneum, the two species which have been formerly described as specific chickpea symbionts. This is the first report on characterization of chickpea nodulating rhizobia covering soils of both northern and peninsular India. The collection of isolates, diverse in terms of species and symbiotic effectiveness holds a vast pool of genetic material which can be effectively used to yield superior inoculant strains.     

Fitness variation among host species and the paradox of ineffective rhizobia

Legumes can preferentially select beneficial rhizobial symbionts and sanction ineffective strains that fail to fix nitrogen. Yet paradoxically, rhizobial populations vary from highly beneficial to ineffective in natural and agricultural soils. Classic models of symbiosis focus on the single dimension of symbiont cost‐benefit to sympatric hosts, but fail to explain the widespread persistence of ineffective rhizobia. Here, we test a novel framework predicting that spatio‐temporal and community dynamics can maintain ineffective strains in rhizobial populations. We used clonal and multistrain inoculations and quantitative culturing to investigate the relative fitness of four focal Bradyrhizobium strains varying from effective to ineffective on Acmispon strigosus. We found that an ineffective Bradyrhizobium strain can be sanctioned by its native A. strigosus host across the host's range, forming fewer and smaller nodules compared to beneficial strains. But the same ineffective Bradyrhizobium strain exhibits a nearly opposite pattern on the broadly sympatric host Acmispon wrangelianus, forming large nodules in both clonal and multistrain inoculations. These data suggest that community‐level effects could favour the persistence of ineffective rhizobia and contribute to variation in symbiotic nitrogen fixation.     

Salt- and alkaline-tolerance are linked in Acacia

Saline or alkaline soils present a strong stress on plants that together may be even more deleterious than alone. Australia''s soils are old and contain large, sometimes overlapping, areas of high salt and alkalinity. Acacia and other Australian plant lineages have evolved in this stressful soil environment and present an opportunity to understand the evolution of salt and alkalinity tolerance. We investigate this evolution by predicting the average soil salinity and pH for 503 Acacia species and mapping the response onto a maximum-likelihood phylogeny. We find that salinity and alkalinity tolerance have evolved repeatedly and often together over 25 Ma of the Acacia radiation in Australia. Geographically restricted species are often tolerant of extreme conditions. Distantly related species are sympatric in the most extreme soil environments, suggesting lack of niche saturation. There is strong evidence that many Acacia have distributions affected by salinity and alkalinity and that preference is lineage specific.     

Origins of cheating and loss of symbiosis in wild Bradyrhizobium

Rhizobial bacteria nodulate legume roots and fix nitrogen in exchange for photosynthates. These symbionts are infectiously acquired from the environment and in such cases selection models predict evolutionary spread of uncooperative mutants. Uncooperative rhizobia – including nonfixing and non‐nodulating strains – appear common in agriculture, yet their population biology and origins remain unknown in natural soils. Here, a phylogenetically broad sample of 62 wild‐collected rhizobial isolates was experimentally inoculated onto Lotus strigosus to assess their nodulation ability and effects on host growth. A cheater strain was discovered that proliferated in host tissue while offering no benefit; its fitness was superior to that of beneficial strains. Phylogenetic reconstruction of Bradyrhizobium rDNA and transmissible symbiosis‐island loci suggest that the cheater evolved via symbiotic gene transfer. Many strains were also identified that failed to nodulate L. strigosus, and it appears that nodulation ability on this host has been recurrently lost in the symbiont population. This is the first study to reveal the adaptive nature of rhizobial cheating and to trace the evolutionary origins of uncooperative rhizobial mutants.     

Effect of salt stress on nodulation and nitrogen fixation in legumes

It is now well established that almost all phases of root nodule development in legumes are adversely affected by saline conditions in the rooting medium. There is also a general agreement that the rhizobia are more tolerant to salt stress than the host plant, but they show considerable strain variability in growth and survival under saline conditions. Inhibitory effect of salinity on nodulation has been attributed to decrease in rhizobial colonisation and shrinkage and lack of root hair formation. Salt stress also induces premature senescence of already formed nodules. Both N2-fixation activity and nodule respiration are inhibited sharply on exposure of plants to saline conditions. The decrease in N2-fixation has been ascribed to direct effect on nitrogenase activity or an indirect effect through decrease in leghemoglobin content, respiratory rate, malate concentrations in nodules and photosynthate availability. Salinity increases oxygen diffusion resistance in the nodules and alters their ultrastructure. Decrease in N2-fixation in nodules under salinity is also accompanied by parallel decrease in the activity of H2O2-scavenging enzymes like catalase, ascorbate peroxidase and the level of antioxidants like ascorbic acid. Nodules appear to undergo osmoregulation under saline conditions by accumulating physiologically compatible solutes like proline, sugars (pinnitol) and lactic acid. The intensity of the adverse effects of salinity on nodule functioning depends on plant species, rhizobial strain, duration of exposure to saline conditions, nature, concentration and mode of salt application.     

Oil content and lipid composition of safflower (Carthamus tinctorius) irrigated with saline water under greenhouse and field conditions

An intensive process of land deterioration of some regions in Uzbekistan including the Aral Sea basin has led to a significant increase in soil salinity levels and consequently to a considerable reduction of total fertile soil area, as these lands are of little use for plant growth. The area is estimated to be more than 1.4 million ha of seabed. As a result, there was an immediate need to cultivate new crops capable of stopping the movement of sands and the enlargement of saline soils. Safflower (Carthamus tinctorius) is considered to be a moderately salt‐tolerant crop and, as such, one of a few crops well suited to the cropping systems of salt‐affected soils. It is used in Uzbekistan as a reserve crop when the culture of the main crop fails. In spite of the great economic importance of this oil, there is almost no available information in the literature on the effect of salinity on oil quality and its chemical ingredients. The purpose of the present study was to determine, in greenhouse and field experiments, how irrigation with saline water would influence content of oil, lipids and other lipophylic components in safflower. We found that irrigation of safflower with moderate concentrations of saline water seems feasible, as far as oil and lipid composition is concerned. Consequently, safflower might be a potential crop for lands of little use for plant growth in Uzbekistan or other similar sites in the world.     

Effects of peanut rhizobia on the growth and symbiotic performance ofArachis hypogaea under abiotic stress

The effects of saline and osmotic stress on four peanut rhizobia, plant growth and symbiotic N₂-fixation inArachis hypogaea were studied. Abiotic stress was applied by adding either 100 mM NaCl or 20 mM PEG6000. At the rhizobial level,Bradyrhizobium ATCC10317 and TAL1000 showed stronger tolerance to stress than TAL1371 and SEMIA6144. The effect of salinity on the bacterium-plant association was studied by using the variety Blanco Manfredi M68. In the absence of stresses, all the strains induced a significantly higher number of nodules on the roots, although TAL1371 and SEMIA6144 were more effective. Both stresses affected the interaction process, while TALl371 was the best partner.     

Microbial phylotype composition and diversity predicts plant productivity and plant–soil feedbacks

The relationship between ecological variation and microbial genetic composition is critical to understanding microbial influence on community and ecosystem function. In glasshouse trials using nine native legume species and 40 rhizobial strains, we find that bacterial rRNA phylotype accounts for 68% of amoung isolate variability in symbiotic effectiveness and 79% of host specificity in growth response. We also find that rhizobial phylotype diversity and composition of soils collected from a geographical breadth of sites explains the growth responses of two acacia species. Positive soil microbial feedback between the two acacia hosts was largely driven by changes in diversity of rhizobia. Greater rhizobial diversity accumulated in association with the less responsive host species, Acacia salicina, and negatively affected the growth of the more responsive Acacia stenophylla. Together, this work demonstrates correspondence of phylotype with microbial function, and demonstrates that the dynamics of rhizobia on host species can feed back on plant population performance.     

Phenotypic plasticity with respect to salt stress response by Lotus glaber: the role of its AM fungal and rhizobial symbionts

Our hypothesis is that Lotus glaber (a glycophytic species, highly tolerant to saline-alkaline soils) displays a plastic root phenotypic response to soil salinity that may be influenced by mycorrhizal and rhizobial microorganisms. Uninoculated plants and plants colonised by Glomus intraradices or Mesorhizobium loti were exposed to either 150 or 0 mM NaCl. General plant growth and root architectural parameters (morphology and topology) were measured and phenotypic plasticity determined at the end of the salt treatment period. Two genotypes differing in their salt tolerance capacity were used in this study. G. intraradices and M. loti reduced the total biomass of non-salinised, sensitive plants, but they did not affect that of corresponding tolerant ones. Root morphology of sensitive plants was greatly affected by salinity, whereas mycorrhiza establishment counteracted salinity effects. Under both saline conditions, the external link length and the internal link length of mycorrhizal salt-sensitive plants were higher than those of uninoculated control and rhizobial treatments. The topological trend (TT) was strongly influenced by genotype x symbiosis interaction. Under non-saline conditions, nodulated root systems of the sensitive plant genotype had a more herringbone architecture than corresponding uninoculated ones. At 150 mM NaCl, nodulated root systems of tolerant plants were more dichotomous and those of the corresponding sensitive genotype more herringbone in architecture. Notwithstanding the absence of a link between TTs and variations in plant growth, it is possible to predict a dissimilar adaptation of plants with different TTs. Root colonisation by either symbiotic microorganisms reduced the level of root phenotypic plasticity in the sensitive plant genotype. We conclude that root plasticity could be part of the general mechanism of L. glaber salt tolerance only in the case of non-symbiotic plants.     

Genetic diversity of <Emphasis Type="Italic">Sinorhizobium meliloti</Emphasis> associated with alfalfa in Chilean volcanic soils and their symbiotic effectiveness under acidic conditions

A study was conducted with the aim of evaluating the genetic diversity of alfalfa rhizobia isolated from volcanic soils in southern Chile and their ability to establish an effective symbiosis with alfalfa. Rhizobial strains isolated from nodules were identified and selected based on PCR analyses and acid tolerance. Symbiotic effectiveness (nodulation and shoot dry weight) of acid-tolerant rhizobia was evaluated in glasshouse experiments under acidic conditions. The results revealed that Sinorhizobium meliloti is the dominant species in alfalfa nodules with a high genetic diversity at strain level grouped in three major clusters. There was a close relationship (r ² = 0.895, P ≤ 0.001, n = 40) between soil pH and the size of rhizobial populations. Representative isolates from major cluster groups showed wide variation in acid tolerance expressed on buffered agar plates (pH 4.5–7.0) and symbiotic effectiveness with alfalfa. One isolate (NS11) appears to be suitable as an inoculant for alfalfa according to its acid tolerance and symbiotic effectiveness at low pH (5.5). The isolation and selection of naturalized S. meliloti strains with high symbiotic effectiveness under acidic conditions is an alternative approach to improving the productivity of alfalfa and for reducing the application of synthetic fertilizers in Chile.     

Genetic Diversity of <Emphasis Type="Italic">Acacia seyal</Emphasis> Del. Rhizobial Populations Indigenous to Senegalese Soils in Relation to Salinity and pH of the Sampling Sites

The occurrence and the distribution of rhizobial populations naturally associated to Acacia seyal Del. were characterized in 42 soils from Senegal. The diversity of rhizobial genotypes, as characterized by polymerase chain reaction restriction fragment length polymorphism (RFLP) analysis of 16S–23S rDNA, performed on DNA extracted from 138 nodules resulted in 15 clusters. Results indicated the widespread occurrence of compatible rhizobia associated to A. seyal in various ecogeographic areas. However, the clustering of rhizobial populations based on intergenic spacer (IGS) RFLP profiles did not reflect their geographic origin. Four genera were discriminated on the basis of 16S rRNA gene sequences of the strains representative for the IGS-RFLP profiles. The majority of rhizobia associated to A. seyal were affiliated to Mesorhizobium and Sinorhizobium 64 and 29%, respectively, of the different IGS-RFLP profiles. Our results demonstrate the coexistence inside the nodule of plant-pathogenic non-N₂-fixing Agrobacterium and Burkholderia strains, which induced the formation of ineffective nodules, with symbiotic rhizobia. Nodulation was recorded in saline soils and/or at low pH values or in alkaline soils, suggesting adaptability of natural rhizobial populations to major ecological environmental stress and their ability to establish symbiotic associations within these soil environments. These results contribute to the progressing research efforts to uncover the biodiversity of rhizobia and to improve nitrogen fixation in agroforestry systems in sub-Saharan Africa.     

Comparison of Distichlis spicata and Suaeda aegyptiaca in response to water salinity: Candidate halophytic species for saline soils remediation

Distichlis spicata and Suaeda aegyptiaca are two potential halophytic plant species for bioremediation of salt degraded soils, and development of saline agriculture. The physiological responses of the species to different levels of salinity (EC 0, 12, 24, 36, and 48 dS/m) in a controlled environment experiment were studied. Both species showed a high level of tolerance to elevated concentrations of salt in the irrigation water. The shoot fresh and dry weights in S. aegyptiaca increased till 36 dS/m and were sustained under 48 dS/m while in D. spicata, both parameters decreased as salinity increased. Glycine betaine accumulation did not change in D. spicata with increasing salinity, whereas proline content revealed a marked increase of 7.13 fold in 48 dS/m salinity compared to the control, which showed its critical osmoprotection role in the plant. In S. aegyptiaca, both osmolytes content significantly increased at high salinity levels (36 and 48 dS/m) up to 3.22 and 2.0 folds, respectively. Overall, S. aegyptiaca had a better potential of Na⁺ phytoremediation, and tolerated higher salinity compared to D. spicata. In contrast, the vigorous root and rhizome growth in D. spicata made it a proper solution for protecting the soils against further erosion under saline conditions.     

Diversity and Biogeography of Rhizobia Isolated from Root Nodules of <Emphasis Type="Italic">Glycine max</Emphasis> Grown in Hebei Province,China

A total of 215 rhizobial strains were isolated and analyzed with 16S rRNA gene, 16S–23S intergenic spacer, housekeeping genes atpD, recA, and glnII, and symbiotic genes nifH and nodC to understand the genetic diversity of soybean rhizobia in Hebei province, China. All the strains except one were symbiotic bacteria classified into nine genospecies in the genera of Bradyrhizobium and Sinorhizobium. Surveys on the distribution of these rhizobia in different regions showed that Bradyrhizobium japonicum and Bradyrhizobium elkanii strains were found only in neutral to slightly alkaline soils whereas Bradyrhizobium yuanmingense, Bradyrhizobium liaoningense-related strains and strains of five Sinorhizobium genospecies were found in alkaline–saline soils. Correspondence and canonical correspondence analyses on the relationship of rhizobial distribution and their soil characteristics reveal that high soil pH, electrical conductivity, and potassium content favor distribution of the B. yuanmingense and the five Sinorhizobium species but inhibit B. japonicum and B. elkanii. High contents of available phosphorus and organic matters benefit Sinorhizobium fredii and B. liaoningense-related strains and inhibit the others groups mentioned above. The symbiotic gene (nifH and nodC) lineages among B. elkanii, B. japonicum, B. yuanmingense, and Sinorhizobium spp. were observed in the strains, signifying that vertical gene transfer was the main mechanism to maintain these genes in the soybean rhizobia. However, lateral transfer of symbiotic genes commonly in Sinorhizobium spp. and rarely in Bradyrhizobium spp. was also detected. These results showed the genetic diversity, the biogeography, and the soil determinant factors of soybean rhizobia in Hebei province of China.     

Importance of native arbuscular mycorrhizal inoculation in the halophyte Asteriscus maritimus for successful establishment and growth under saline conditions

Background and aims

The biological restoration of saline habitats could be achieved by using halophyte plant species together with adapted arbuscular mycorrhizal fungi (AMF). An interesting plant to be used in restoration of saline environments, Asteriscus maritimus, is highly mycotrophic. The aim of this study was to assess the effectiveness of native and allochthonous AMF to enhance the establishment and growth of the halophyte A. maritimus under saline conditions.

Methods

We studied the symbiotic effectiveness of four AMF strains (three native fungal isolates from a saline soil and one allochthonous, from collection) in A. maritimus subjected to increasing salinity stress. We measured plant physiological parameters by which AMF may ameliorate the detrimental effects of salinity stress.

Results

A. maritimus plants showed a high mycorrhizal dependency, even in absence of salt stress. Plants inoculated with native AMF had higher shoot dry weight, efficiency of photosystem II, stomatal conductance and accumulation of glutathione than those inoculated with the collection AMF at the highest level of salinity. Moreover, at this salt level, only 30 % of A. maritimus plants inoculated with the collection AMF survived, while with the three native AMF, the rate of survival was 100 %.

Conclusions

Results points out the importance of native AMF inoculation in the establishment, survival and growth of A. maritimus plants. Inoculation with these native AMF enhanced A. maritimus salt tolerance by increasing efficiency of photosystem II, stomatal conductance and glutathione content and by reducing oxidative damage. Thus, the use of adequate native AMF inocula could be a critical issue for success in recovering saline degraded areas.     

Characterization of high temperature-tolerant rhizobia isolated from Prosopis juliflora grown in alkaline soil

A method was developed for the fast screening and selection of high-temperature tolerant rhizobial strains from root nodules of Prosopis juliflora growing in alkaline soils. The high-temperature tolerant rhizobia were selected from 2,500 Rhizobium isolates with similar growth patterns on yeast mannitol agar plates after 72 h incubation at 30 and 45 degrees C, followed by a second screening at 47.5 degrees C. Seventeen high-temperature tolerant rhizobial strains having distinguishable protein band patterns were finally selected for further screening by subjecting them to temperature stress up to 60 degrees C in yeast mannitol broth for 6 h. The high-temperature tolerant strains were NBRI12, NBRI329, NBRI330, NBRI332, and NBRI133. Using this procedure, a large number of rhizobia from root nodules of P. juliflora were screened for high-temperature tolerance. The assimilation of several carbon sources, tolerance to high pH and salt stress, and ability to nodulate P. juliflora growing in a glasshouse and nursery of the strains were studied. All five isolates had higher plant dry weight in the range of 29.9 to 88.6% in comparison with uninoculated nursery-grown plants. It was demonstrated that it is possible to screen in nature for superior rhizobia exemplified by the isolation of temperature-tolerant strains, which established effective symbiosis with nursery-grown P. juliflora. These findings indicate a correlation between strain performance under in vitro stress in pure culture and strain behavior under symbiotic conditions. Pure culture evaluation may be a useful tool in search for Rhizobium strains better suited for soil environments where high temperature, pH, and salt stress constitutes a limitation for symbiotic biological nitrogen fixation.     

Genotypic and phenotypic characterization of rhizobia that nodulate snap bean (Phaseolus vulgaris L.) in Egyptian soils

Snap bean fields in 12 of the 25 governorates of Egypt were surveyed to determine the distribution and taxonomy of snap bean-nodulating rhizobia. Nodulation rates in the field were very low, indicating that Egyptian soils do not have sufficient numbers of snap bean-compatible Rhizobium spp. A total of 87 rhizobial isolates were assayed on the most commonly grown cultivars in order to identify the most effective strains. The five most effective isolates (R11, R13, R28, R49 and R52) were fast-growing and utilized a wide range of carbon and nitrogen sources. A phylogenetic assignment of these strains by analysis of the 16S ribosomal RNA gene suggested that all fell within the Rhizobium etli–Rhizobium leguminosarum group. Strains R11, R49 and R52 all clustered with other identified R. etli strains, while strains R13 and R28 were more distinct. The distinctness of R13 and R28 was supported by physiological characteristics, such as their ability to utilize citrate, erythritol, dulcitol and lactate. Strains R13 and R28 also yielded the highest plant nitrogen content of all isolates.The highly effective strains isolated in this study, in particular strains R13 and R28, are promising candidates for improving crop yields. The data also suggested that these two strains represented a novel sub-group within the R. etli–R. leguminosarum group. As snap bean is a crop of great economic value to Egypt, the identification of highly effective rhizobial strains adapted to Egyptian soils, such as strains R13 and R28, is of great interest.