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The genetic diversity and phylogeny of rhizobia isolated from nodules of 9 wild-growing Lathyrus L. species (Fabaceae) growing in Republic Bashkortostan were studied. It is shown that for the given plants is characteristic that the big variety of heterogeneous strains of root nodule bacteria. Nevertheless, it is revealed that the majority of them in phylogenetics are closely related to Rhizobium leguminosarum. However, some plant species are found out also nodule bacteria which were considered earlier unusual for Lathyrus. So, L. vernus L. Bernh. and L. sylvestris L. are found out a root nodule bacteria close to R. tropici, L. palustris L. — Agrobacterium sp., and L. gmelinii Fritsch all isolated with us bacteria from root nodules by the sequence of genes of 16S рРНК have appeared are closely related to Phyllobacterium myrsinacearum.

The Sinorhizobium-legume interaction is sensitive to a number of environmental factors: one of the important stresses is soil acidity. Low pH imposes severe constraints on legume productivity. In the Sinorhizobium-Medicago partnership, prokaryotic partner is the most acid-sensitive. The isolation of acid-tolerant Sinorhizobium strains from the eastern and southern Mediterranean rim have allowed the successful development of medic pastures on over I million ha of land in Western Australia, posing two important questions: • How do acid-tolerant strains differ from acid-sensitive counterparts? • How do agriculturally important root nodule bacteria respond to low pH? Knowledge of the genetic circuits required and activated by the cell in acidic conditions is vital to understand the response and survival of bacteria in these conditions. The molecular basis of acid tolerance has been investigated with several approaches (Glenn et al. 1999).

Quantification of acidity tolerance in the laboratory may be the first step in rhizobial strain selection for the Amazon region. The present method evaluated rhizobia in Petri dishes with YMA medium at pH 6.5 (control) and 4.5, using scores of 1.0 (sensitive, "no visible" growth) to 4.0 (tolerant, maximum growth). Growth evaluations were done at 6, 9, 12, 15 and 18 day periods. This method permits preliminary selection of root nodule bacteria from Amazonian soils with statistical precision. Among the 31 rhizobia strains initially tested, the INPA strains 048, 078, and 671 presented scores of 4.0 at both pHs after 9 days of growth. Strain analyses using a less rigorous criterion (growth scores higher than 3.0) included in this highly tolerant group the INPA strains 511, 565, 576, 632, 649, and 658, which grew on the most diluted zone (zone 4) after 9 days. Tolerant strains still must be tested for nitrogen fixation effectiveness, competitiveness for nodule sites, and soil persistence before their recommendation as inoculants.

International audience; The aim of this study was to identify bacteria present in the nodules of the legume Hedysarum spinosissimum growing in metal-contaminated soils; and to test whether these root-nodule bacteria are able to promote host plant growth and enhance their phytostabilization potential. Seventy-four bacteria were isolated from nodules of H. spinosissimum growing in 3 different mining sites in Morocco and were identified by 16S rDNA gene sequencing. They belonged to 8 genera affiliated to Pseudomonas (49 strains), Pantoea (11), Rhizobium (6), Herbaspirillum (3), Bacillus (2), and one strain of Serratia, Agrobacterium and Azospirillum. Seven strains, presenting a high tolerance to Zn and Pb, exhibited capacity of inorganic phosphate solubilization and ammonia production from peptone degradation. The inoculation of H. spinosissimum, growing in 2000 μM of Zn by Pseudomonas putida and Herbaspirillum huttiense suppresses Zn toxicity symptoms in plants and also enhances plant growth by significantly increasing plant shoot and root fresh weights. The maximal Zn accumulation was observed in roots of plants inoculated by Pseudomonas putida with a translocation factor of 0.05(\textpm 0.006). Our results, evidence that the selection of metal-resistant bacteria is a key step in polluted soils for the use of plants like Hedysarum spinosissimum for in situ phytoremediation.

In the present study we recovered endophytic root-nodule bacteria from chickpea (Cicer arietinumi L.) and mothbean (Vigna aconitifolia L.). Phenotypic and genotypic characterization of isolates was performed by employing biochemical and genetic approaches. Sequencing data showed that most isolates belonged to genus, Pseudomonas spp. being a dominant species. They also showed similarity with Rhizobium, Agrobacterium and Erwinia spp. Isolates were screened functionally for indole-3-acetic acid, siderophore production and inorganic phosphorus (Pi) solubilization. All isolates showed Pi solubilization except CJS-2. Nine isolates (CSS-1, CBS-1, CLS-3, CCS-1, CHS-1, VS-1, VL-1, VN-1, VN-2) were found positive for IAA production and eight isolates (CBS-1, CCS-1, CHS-2, CKS-2, CNS-2, VS-1, VJ-1) exhibited positive results for siderophore production. An understanding of the phonetic and functional diversity of these microbes that interact with plants will be worthwhile to fully achieve the biotechnological potential of efficient plant-microbe partnerships for a range of applications.

Inoculation with beneficial soil microorganisms has the potential to enhance success of restoration, particularly in harsh Mediterranean-type ecosystems (MTEs). We investigated the effects of microorganisms (mycorrhizal fungi and root nodule bacteria) and planting material (seed and nursery-raised seedlings) on early establishment and growth of two key postdisturbance colonizing species with different life histories, life forms and functional types (Eucalyptus gomphocephala and Acacia saligna) under field conditions. Establishment and growth were monitored at 13months, following the first MTE drought period. For E. gomphocephala, establishment was higher for seedlings (81%) than for seeding (7.5%). Inoculation with ectomycorrhizal fungal spores was not beneficial. For A. saligna, establishment was also higher for seedlings (84%) than for seeding (42.5%). Mycorrhizal fungal inoculum had no effect on establishment or growth. This study has shown that in harsh MTE conditions, the use of seedlings is more effective than seeding in degraded woodlands even when attempting to reintroduce key colonizing species. The microorganism treatments tested did not result in significant improvement in establishment or growth.

Very little is known about the genetic diversity and phylogeny of rhizobia nodulating Lotus species in northern temperate regions. We have therefore studied the genetic diversity among a total of 61 root nodule bacteria isolated from Lotus corniculatus and Anthyllis vulneraria from different geographic sites and habitats in Sweden by restriction fragment length polymorphism (RFLP) of the internal transcribed spacer between their 16S rRNA and 23S rRNA (IGS) region. A high diversity consisting of 26 IGS types from 54 L. corniculatus isolates and five IGS types from seven A. vulneraria isolates was found. The 16S rRNA sequences and phylogeny of representatives of the different IGS types showed four interesting exceptions from the majority of the isolates belonging to the genus Mesorhizobium: Two isolates were both found to be closely related to Rhodococcus spp., and two other isolates showed close relationship with Geobacillus spp. and Paenibacillus spp., respectively. The nodA sequences and phylogeny showed that all the isolates, including those not belonging to the traditional rhizobia genera, harbored nodA sequences which were typical of Mesorhizobium loti. Generally, the 16S rRNA and nodA phylogenetic trees were not congruent in that isolates with similar 16S rRNA sequences were associated with isolates harboring different nodA sequences. All the isolates were confirmed to nodulate L. corniculatus in an inoculation test. This is the first report of members of these non-rhizobia genera being able to nodulate legumes, and we suggest that they may have acquired their nodulating properties through lateral gene transfer.

Liquid media containing potato extract and 1% of glucose or sucrose were used to culture root-nodule bacteria (rhizobia) in shaken Erlenmeyer flasks. For comparison, these bacteria were also cultured in yeast extract-mannitol broth (YEMB) as a standard medium. Proliferation of rhizobia was monitored by measuring optical densities (OD550) of the cultures and by plate counting of the viable cells (c.f.u) of the bacteria. In general, multiplication of the rhizobia in potato extract-glucose broth (PEGB) and potato extract-sucrose broth (PESB) was markedly faster, as indicated by higher values of OD550, than in YEMB. The numbers of R. leguminosarum by. vicae GGL and S. meliloti 330 in PEGB and PEGB were high and ranged from 1.2 x 10(10) to 4.9 x 10(10) mL(-1) after 48 h of incubation at 28 degrees C. B. japonicum B3S culture in PEGB contained 6.4 x 10(9) c.f.u. ml(-1) after 72 h of incubation. PEGB and YEMB cultures of the rhizobia were similar with respect to their beneficial effects on nodulation of the host-plants of these bacteria.

Native s-glucuronidase activity is absent in the root nodule bacteria. The Escherichia coli gusA gene (conferring s-glucuronidase activity) can be used as a tool for the genetic analysis of these bacteria. The gusA was modified by positioning stop codons in all three reading frames (before the RBS of gusA) to prevent translational fusions.

Plant growth experiments were conducted to assess symbiotic efficiency, photosynthetic rates, and the development of soybean (Glycine max (L.) Merrill) seedlings after seed inoculation with active and inactive strains of root nodule bacteria Bradyrhizobium japonicum preincubated in the presence homologous and heterologous proteins. The properties of active and inactive symbiotic strains were differentially modulated by homologous soybean lectin, which had a marked influence on plant physiological condition. The incubation of active rhizobia with a homologous lectin, i.e., lectin of the respective plant, increased the nitrogen-fixing activity of nodules and, consequently, elevated photosynthetic rates and weight increments in soybean plants. At the same time, the homologous lectin suppressed the symbiotic properties of inactive strain of nodule bacteria. The preincubation of rhizobia with a heterologous pea lectin had virtually no effect on functioning of symbiotic apparatus and photosynthetic rate, whereas the preincubation of root nodule bacteria with human albumin exerted an effect similar to that induced by a homologous lectin on symbiotic productivity.

© 2015, Nature Publishing Group. All rights reserved. Root nodule bacteria (RNB) or "rhizobia" are a type of plant growth promoting bacteria, typified by their ability to fix nitrogen for their plant host, fixing nearly 65% of the nitrogen currently utilized in sustainable agricultural production of legume crops and pastures. In this study, we sequenced the genomes of 110 RNB from diverse hosts and biogeographical regions, and undertook a global exploration of all available RNB genera with the aim of identifying novel genetic determinants of symbiotic association and plant growth promotion. Specifically, we performed a subtractive comparative analysis with non-RNB genomes, employed relevant transcriptomic data, and leveraged phylogenetic distribution patterns and sequence signatures based on known precepts of symbiotic- and host-microbe interactions. A total of 184 protein families were delineated, including known factors for nodulation and nitrogen fixation, and candidates with previously unexplored functions, for which a role in host-interaction, -regulation, biocontrol, and more, could be posited. These analyses expand our knowledge of the RNB purview and provide novel targets for strain improvement in the ultimate quest to enhance plant productivity and agricultural sustainability.

This study was conducted to select rhizobia from hairy vetch (Vicia villosa Roth) with nodulation and excellent nitrogen-fixing ability. Hairy vetch root was collected from 7 of cultivation region of all over the country, rhizobia were isolated from the root nodules. Isolates were re-inoculated into a hairy vetch separately and studied nodulation and nitrogen-fixing ability. As a result, total of 52 Rhizobium isolates were isolated from the hairy vetch root nodules, among these, 16 isolates were Rhizobium which show good growth at more than 0.5% NaCl concentration. These 16 isolates were re-inoculated separately, 8 weeks after, good root nodule formation was observed from Rhizobium sp. RH1, RH3, RH81, RH82, RH84, and RH93 strain treated samples. Six isolates were positive for nitrogen fixing ability, the highest acetylene reduction activity was shown by Rhizobium sp. RH84. Results suggest that the Rhizobium sp. RH84 could be used as the possibility of its application as a green manure crop of hairy vetches in nonuniform salt distribution reclaimed land.

A collection of 104 isolates from root-nodules of Vicia faba was submitted to 16S rRNA PCR-RFLP typing. A representative sample was further submitted to sequence analysis of 16S rRNA. Isolates were assigned to 12 genera. All the nodulating isolates (45 %) were closely related to Rhizobium leguminosarum USDA2370(T) (99.34 %). The remaining isolates, including potential human pathogens, failed to nodulate their original host. They were checked for presence of symbiotic genes, P-solubilization, phytohormone and siderophore production, and then tested for their growth promoting abilities. Results indicated that 9 strains could induce significant increase (41-71 %) in shoot dry yield of faba bean. A Pseudomonas strain was further assessed in on-farm trial in combination with a selected rhizobial strain. This work indicated that nodule-associated bacteria could be a valuable pool for selection of effective plant growth promoting isolates. Nevertheless, the possible involvement of nodules in increasing risks related to pathogenic bacteria should not be neglected and needs to be investigated further.

Fourteen root-nodule bacteria isolated from the medicinal legume Indigofera tinctoria were characterized for their phenotypic features including growth curves, utilization of carbon and nitrogen sources, antibiotic resistance, vitamin requirement and growth under different conditions. The partial sequences of the 16S rDNA of these strains were obtained and BLASTN analysis revealed that the microsymbionts of I. tinctoria were related to members of five distinct genera: Rhizobium, Sinorhizobium, Bradyrhizobium, Cupriavidus and Pseudoalteromonas. The partial nifH gene of Pseudoalteromonas-like strain DASA 57075 had 96% similarity with nifH genes of members of Bradyrhizobium. The partial nodC gene of Pseudoalteromonas-like strain DASA 57075 showed 88% similarity with the nodC gene of several rhizobia including Sinorhizobium, Bradyrhizobium and Mesorhizobium. We propose a bacterium that is related to Pseudoalteromonas from the gamma-class of Proteobacteria as a new legume symbiont. This is also the first report that the same species of legume can be nodulated by bacteria from up to five different genera in three distinct classes.

This study aims to evaluate the phenotypical characteristics of bacterial isolates from mulungu (Erythrina velutina Willd.) nodules and determinate their Box-PCR fingerprinting. All bacteria were evaluated by the following phenotypic characteristics: growth rate, pH change, colony color and mucus production. The bacterial isolates able to re-nodulate the original host were also evaluated regarding its tolerance to increased salinity and different incubation temperatures, ability to growth using different carbon sources, intrinsic antibiotic resistance and ?in vitro? auxin biosynthesis. The molecular fingerprints were set up using the Box-PCR technique and the isolates were clustered by their profiles. Among the 22 bacterial isolates obtained, eight were able to re-nodulate the original host. Among the nodule inducing isolates, some were tolerant to 1% of NaCl and 39° C and all of them metabolized the maltose, fructose, glucose, sucrose and arabinose, were resistant to rifampicin and produced auxin. The bacteria showed low genetic similarity among them and reference strains, which indicates the great genetic variability of the isolates. The results of this work are the first reports about the bacterial isolates able to nodulate this species. A more deep study of these bacteria may reveal the existence of isolates tolerant to environmental stresses and suitable as a future mulungu inoculant.

The genus Psoralea (tribe Psoraleae, family Leguminosae) is indigenous to the Cape fynbos of South Africa and little is known about its symbiosis and/or adaptation. The aim of this study was to assess root nodulation and N2 fixation in eight of the 50 Psoralea species, as well as the biodiversity of their associated nodulating microsymbionts. The eight species studied (namely, Psoralea pinnata, Psoralea aphylla, Psoralea aculeata, Psoralea monophylla, Psoralea repens, Psoralea laxa, Psoralea asarina and Psoralea restioides) all had round-shaped, determinate type (desmodioid) nodules, and data from 15N natural abundance showed that they obtained 60–88% of their N nutrition from symbiotic fixation. These Psoralea species also transported their fixed-N as ureides (allantoin and allantoic acid) in the xylem stream, a symbiotic trait that links them very closely to the tribe Phaseoleae. Bacteria isolated from root nodules of the eight Psoralea species varied in phenotypic characteristics, nodulation promiscuity, and N2-fixing efficacy. Furthermore, 16S rDNA gene sequence data showed that Psoralea species can form root nodules with different soil bacteria, including Rhizobium, Mesorhizobium and Burkholderia strains. This is not only evidence of nodulation promiscuity, but also an indication of the species’ adaptation to the nutrient-poor, low-N, sandy acidic soils of the Cape fynbos.

Milletia pinnata is a leguminous tropical tree that produces seed oil suitable for biodiesel and is targeted to be planted on marginal land associated with nitrogen poor soil. This study aimed to identify effective rhizobia species for M. pinnata. Soil samples were collected from M. pinnata grown in Kununurra, Australia. Rhizobia were trapped, characterised and sequenced for 16S rRNA, atpD, dnaK and recA genes. Forty isolates tolerated pH 7 – 9, temperatures 29 – 37 °C, salinity below 1 % NaCl, and had optimal growth on mannitol, arabinose or glutamate as a single carbon source, a few grew on sucrose and none grew on lactose. Inoculation of isolates increased shoot dry weight of M. pinnata’s seedlings in nitrogen minus media. Slow-growing isolates were closely related to Bradyrhizobium yuanmingense, Bradyrhizobium sp. DOA10, Bradyrhizobium sp. ORS305 and B. liaoningense LMG 18230T. The fast-growing isolates related to Rhizobium sp. 8211, R. miluonense CCBAU 41251T, R miluonense CC-B-L1, Rhizobium sp. CCBAU 51330 and Rhizobium sp. 43015 Millettia pinnata was effectively nodulated by slow-growing isolates related to Bradyrhizobium yuanmingense, Bradyrhizobium sp. DOA10 Bradyrhizobium sp. ORS305, B. liaoningense LMG 18230T and fast-growing isolates related Rhizobium sp. 8211, R. miluonense, Rhizobium sp. CCBAU 51330 and Rhizobium sp. 43015

Aims: Aim of this study is to determine the genetic variation of rhizobia associated with horse gram [Macrotyloma uniflorum (Lam.) Verdc.] plants grown in different regions of Andhra Pradesh, India. Methods and Results: Four representative isolates having most representative characters from the previous characterization were selected for 16S rRNA sequence. The sequences were submitted to the NCBI GenBank and Ribosomal Database Project (RDP). The isolates HGR-4, 6 and 13 showed more than 99% homology between them and they were grouped with Rhizobium reference strains where as the isolate HGR-25 showed 87·1, 87·4 and 87·2% homology with the isolates HGR-4, 6 and 13, respectively, and were grouped with reference strains for Caulobacter. The nodulation ability of these isolates on horse gram was confirmed by inoculation tests. Conclusions: The isolate HGR-25 was identified as Caulobacter isolated from the plants growing in soil samples collected from Khareemnagar district, Andhra Pradesh, India. Inoculation tests revealed that Caulobacter formed nodules on horse gram. It was also confirmed by RDP. Significance and Impact of the Study: This is the first report that a legume was nodulated by a member of the genus Caulobacter, which belongs to the family Caulobacteriaceae in the order Caulobacterales of Alphaproteobacteria.

Symbiotic relationships between legumes and nitrogen-fixing soil micro-organisms are of ecological importance in plant communities worldwide. For example, nutrient-poor Australian soils are often dominated by shrubby legumes (e.g. species of Acacia). However, relatively few studies have quantified patterns of diversity, host-specificity and effectiveness of these ecologically important plant–microbe interactions. In this study, 16S rRNA gene sequence and PCR-RFLP analyses were used to examine bacterial strains isolated from the root nodules of two widespread south-eastern Australian legumes, Acacia salicina and Acacia stenophylla, across nearly 60 sites. The results showed that there was extensive genetic diversity in microbial populations, including a broad range of novel genomic species. While previous studies have suggested that most native Australian legumes nodulate primarily with species of the genus Bradyrhizobium, our results indicate significant associations with members of other root-nodule-forming bacterial genera, including Rhizobium, Ensifer, Mesorhizobium, Burkholderia, Phyllobacterium and Devosia. Genetic analyses also revealed a diverse suite of non-nodulating bacterial endophytes, only a subset of which have been previously recorded. Although the ecological roles of these endosymbionts are not well understood, they may play both direct and indirect roles in promoting plant growth, nodulation and disease suppression.

Competitiveness and genetic variation of the Rhizobium galegae strains from the collection of the All-Russia Institute of Agricultural Microbiology, Russian Academy of Agricultural Sciences, causing nodulation of oriental goat's rue under conditions of Bashkortostan soils (lacking this rhizobial species) were studied. It was demonstrated that of all the tested strains, the strains CIAM 0702 and CIAM 0704, each carrying two megaplasmids of 1500 and 2000 MDa, were the most competitive. RAPD (random amplified polymorphic DNA) analysis showed that R. galegae strains were able to intensively exchange the genetic material in the host plant rhizosphere. We did not succeed in detecting the local root nodule bacteria that were either initially able to infect oriental goat's rue or had adapted to infecting this species due to various genetic rearrangements.

The theoretical and experimental data on salt tolerance of root nodule bacteria Sinorhizobium meliloti (Ensifer meliloti), an alfalfa symbiont, and on genetic determination of this feature are reviewed. Extensive data on the genes affecting adaptation of proteobacteria are provided, as well as on the groups of genes with activity depending on the osmolarity of the medium. Structural and functional polymorphism of the bet genes involved in betaine synthesis and transport in S. meliloti is discussed. The phenotypic and. genotypic polymorphism in 282 environmental rhizobial strains isolated from the centers of alfalfa diversity affected by aridity and salinity is discussed. The isolates from the Aral Sea area and northern Caucasus were shown to possess the betC gene represented by two types of alleles: the dominant A-type allele found in Rm 1021 and the less common divergent E-type allele, which was revealed in regions at the frequencies at the frequencies of 0.35 and 0.48, respectively. In the isolates with the salt-tolerant phenotype, which were isolated from root nodules and subsequently formed less effective symbioses with alfalfa, the frequency of E-type alleles was 2.5 times higher. Analysis of the nucleotide and amino acid sequences of the E-type allele of the betC gene revealed that establishment of this allele in the population was a result of positive selection. It is concluded that diversification of the functionally diverse bet genes occurring in S. meliloti affects the salt tolerance and symbiotic effectivity of rhizobia.

The genetic diversity and phylogeny of root module bacteria entering into symbiotic relations with Caragana arborescens introduced on the territory of the Republic of Bashkortostan (RB) were studied. Analysis of the 85 strains isolated from root-nodules showed that, according to the DNA polymorphism revealed by RAPD analysis, they can be divided into 12 groups. It was found that, both in natural habitats and on the territory of the RB, Caragana arborescens is primarily nodulated by Mesorhizobium bacteria whose 16S rRNA gene sequences differ to some degree from those of the bacteria described earlier by Chinese authors. Bacteria assigned to the genus Phyllobacterium based on their 16S rRNA gene sequences were also revealed in plants growing on the territory of the RB.

Some Tetraponera ants (Formicidae, Pseudomyrmecinae) subsist almost entirely on amino acid deficient honeydew secretions of pseudococcids and harbour a dense aggregation of bacterial symbionts in a unique pouch-shaped organ at the junction of the midgut and the intestine. The organ is surrounded by a network of intruding tracheae and Malpighian tubules, suggesting that these bacteria are involved in the oxidative recycling of nitrogen-rich metabolic waste. We have examined the ultrastructure of these bacteria and have amplified, cloned and sequenced ribosomal RNA-encoding genes, showing that the ant pouch contains a series of close relatives of Flavobacteria and Rhizobium, Methylobacterium, Burkholderia and Pseudomonas nitrogen-fixing root-nodule bacteria. We argue that pouch bacteria have been repeatedly 'domesticated' by the ants as nitrogen-recycling endosymbionts. This ant-associated community of mutualists is, to our knowledge, the first finding of symbionts related to root-nodule bacteria in animals.

The effect of various biopreparations of the root nodule bacterium Bradyrhizobium japonicum, modified with a homologous lectin, on the virulence of rhizobia, the nitrogen-fixing activity of root nodules, and the productivity of the soybean (Glycine max (L.) Merr.) was studied. It was shown that a homologous lectin, added to a bacterial suspension when manufacturing biopreparations on a liquid and solid support, increases the efficiency of the soybean symbiotic system and the productivity of the host plant. The potentialities of using bacterial preparations modified with a homologous lectin are discussed.

Thirty root-nodulating bacteria isolates were obtained from the roots of Acacia ampliceps (Maslin), A. ehrenbergiana (Hayne.), A. saligna (Labill.), A. seyal (Del.), A. tortilis (Forssk.), A. tortilis subsp. raddiana (Savi.), Leucaena leucocephala (Lam.) and Vicia faba (L.) trees growing in the Riyadh region. The isolates' phenotypic and biochemical properties were characterized by assessing colony appearance, growth rate, resistance to antibiotics and heavy metals, and tolerance to salinity, elevated temperature and pH. All isolates had same colony morphology and grew on yeast extract mannitol and tryptone yeast agar, but not MGS media. The results also revealed considerable diversity among the isolates, which exhibited different patterns of resistance to abiotic stresses. Most isolates tolerated temperatures up to 37 degrees C and could grow from pH 5.5-8.5 and at a high NaCl concentration (2% w/v). The majority of isolates could utilize a variety of carbohydrates. Most of the isolates displayed resistance to antibiotics in the 75 microg ml(-1) range, with approximately 100 pg ml(-1) the maximum concentration at which growth was observed. All isolates were sensitive to aluminum and resistant to other heavy metals tested, and they were able to reduce nitrate and hydrolyze urea.