Symbiosis
Symbiosis has played a key role in the evolution of life on Earth. Symbiotic mergers of once independent species drove the origin of eukaryotes. Moreover, symbiosis has enabled many species to gain novel functions and occupy new ecological niches, thus underpinning the functioning of diverse ecosystems. As endosymbionts, microbes provide their eukaryotic hosts with an array of ecological and physiological innovations, including new metabolic capabilities, such as autotrophy or nitrogen fixation, and protection against infections or environmental stressors. Microbial eukaryotes also commonly host their own endosymbionts, including bacteria and algae. Understanding the stability and resilience of symbioses is key to predicting the response of important ecosystems, such as coral reefs, to global change. Manipulating symbiotic associations also has far-reaching economic, environmental and medical implications, through the potential to improve crop productivity, reduce reliance on fertilisers, and control the insect vectors of infectious diseases.
This collection, guest edited by Professor Michael Brockhurst (University of Manchester) and Dr. Rebecca J Hall (University of Birmingham), will feature microbe-focused studies of symbiosis, ranging from the molecular mechanisms of host-symbiont interactions, their genetic and genomic diversity, to understanding the impacts of symbioses in natural and manmade ecosystems.
Collection Contents
41 - 60 of 68 results
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Gene expression level influences amino acid usage, but not codon usage, in the tsetse fly endosymbiont Wigglesworthia
More LessWigglesworthia glossinidia brevipalpis, the obligate bacterial endosymbiont of the tsetse fly Glossina brevipalpis, is characterized by extreme genome reduction and AT nucleotide composition bias. Here, multivariate statistical analyses are used to test the hypothesis that mutational bias and genetic drift shape synonymous codon usage and amino acid usage of Wigglesworthia. The results show that synonymous codon usage patterns vary little across the genome and do not distinguish genes of putative high and low expression levels, thus indicating a lack of translational selection. Extreme AT composition bias across the genome also drives relative amino acid usage, but predicted high-expression genes (ribosomal proteins and chaperonins) use GC-rich amino acids more frequently than do low-expression genes. The levels and configuration of amino acid differences between Wigglesworthia and Escherichia coli were compared to test the hypothesis that the relatively GC-rich amino acid profiles of high-expression genes reflect greater amino acid conservation at these loci. This hypothesis is supported by reduced levels of protein divergence at predicted high-expression Wigglesworthia genes and similar configurations of amino acid changes across expression categories. Combined, the results suggest that codon and amino acid usage in the Wigglesworthia genome reflect a strong AT mutational bias and elevated levels of genetic drift, consistent with expected effects of an endosymbiotic lifestyle and repeated population bottlenecks. However, these impacts of mutation and drift are apparently attenuated by selection on amino acid composition at high-expression genes.
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Fur is not the global regulator of iron uptake genes in Rhizobium leguminosarum
More LessRhizobium leguminosarum fur mutants were unaffected in Fe-dependent regulation of several operons that specify different Fe uptake systems, yet cloned R. leguminosarum fur partially corrected an Escherichia coli fur mutant and R. leguminosarum Fur protein bound to canonical fur boxes. The lack of a phenotype in fur mutants is not due to functional redundancy with Irr, another member of the Fur superfamily found in the rhizobia, since irr fur double mutants are also unaffected in Fe-responsive regulation of several operons involved in Fe uptake. Neither Irr nor Fur is needed for symbiotic N2 fixation on peas. As in Bradyrhizobium japonicum, irr mutants accumulated protoporphyrin IX. R. leguminosarum irr is not regulated by Fur and its Irr protein lacks the motif needed for haem-dependent post-translational modification that occurs in B. japonicum Irr. The similarities and differences in the Fur superfamily in the rhizobia and other Gram-negative bacteria are discussed.
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Small genome of Candidatus Blochmannia, the bacterial endosymbiont of Camponotus, implies irreversible specialization to an intracellular lifestyle a
More LessaThe GenBank accession number for the sequence reported in this paper is AF495758.
Blochmannia (Candidatus Blochmannia gen. nov.) is the primary bacterial endosymbiont of the ant genus Camponotus. Like other obligate endosymbionts of insects, Blochmannia occurs exclusively within eukaryotic cells and has experienced long-term vertical transmission through host lineages. In this study, PFGE was used to estimate the genome size of Blochmannia as approximately 800 kb, which is significantly smaller than its free-living relatives in the enterobacteria. This small genome implies that Blochmannia has deleted most of the genetic machinery of related free-living bacteria. Due to restricted gene exchange in obligate endosymbionts, the substantial gene loss in Blochmannia and other insect mutualists may reflect irreversible specialization to a host cellular environment.
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Reassessment of major products of N2 fixation by bacteroids from soybean root nodules
More LessNH3/ was the principal product from soybean bacteroids, prepared by various procedures, when assayed in solution in a flow chamber under N2 fixation conditions. In addition, small quantities of alanine were produced (reaching 20% of NH3/ under some conditions). Some 15N was assimilated by bacteroids purified from soybean root nodules on Percoll density gradients and shaken with 15N2 and 0·008 atm O2. Under these conditions, accounted for 93% of the 15N fixed into the soluble fraction. This fraction contained no measurable [15N]alanine. Neither these bacteroids nor those prepared by the previously used differential centrifugation method, when incubated with exogenous alanine under non-N2-fixing conditions, gave rise to NH3 from alanine. Therefore, contamination of bacteroid preparations with enzymes of plant cytosolic origin and capable of producing NH3 from alanine cannot explain the failure to detect [15N]alanine [as reported elsewhere: Waters, J. K., Hughes, B. L., II, Purcell, L. C., Gerhardt, K. O., Mawhinney, T. P. & Emerich, D. W. (1998). Proc Natl Acad Sci USA 95, 12038–12042]. Cell-free extracts of the bacteroids as used in the 15N experiments contained alanine dehydrogenase and were able to produce alanine from pyruvate and . Other experiments with alanine dehydrogenase in extracts of cultured rhizobia and bacteroids are reported and discussed in relation to the 15N experiments. Possible reasons for the differences between laboratories regarding the role of alanine are discussed. It is concluded that NH3 is the principal soluble product of N2 fixation by suspensions of soybean bacteroids ex planta and that should continue to be considered the principal product of N2 fixation which is assimilated in vivo in soybean nodules.
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Establishment of a functional symbiosis between the cyanobacterium Nostoc punctiforme and the bryophyte Anthoceros punctatus requires genes involved in nitrogen control and initiation of heterocyst differentiation
More LessThe GenBank accession numbers for the coding regions of ntcA, hetR and hetF are AY038370, AF318069 and AF288130, respectively.
Three mutant strains (ntcA, hetR, hetF) of the cyanobacterium Nostoc punctiforme unable to differentiate heterocysts were characterized and examined for their ability to form a symbiotic association with the bryophyte Anthoceros punctatus. Previously unknown characteristics of the N. punctiforme hetR mutant include differentiation of chilling-resistant akinetes, while vegetative cells of the ntcA mutant randomly lysed, yielding short filaments, following ammonium deprivation. Strains with mutations in hetF and hetR infected A. punctatus with similar frequency to that of wild-type N. punctiforme but did not support growth of the plant partner. These results confirm that the infection of A. punctatus by hormogonia leading to the establishment of an association is physiologically uncoupled from the development of a functional diazotrophic association. They also indicate that heterocyst regulatory elements downstream from HetR and HetF are required in both free-living and symbiotic heterocyst differentiation and nitrogenase expression. A strain with a mutation in the global nitrogen regulator ntcA did not infect A. punctatus despite its ability to differentiate hormogonia at a low frequency. When complemented with one or more copies of ntcA, the mutant strain infected A. punctatus at a similar frequency as the wild-type and supported growth of the plant partner in the absence of combined nitrogen. These results established a connection between the presence of a functional copy of ntcA and the magnitude of hormogonium differentiation, and the behaviour of the formed hormogonia.
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Classification of rhizobia based on nodC and nifH gene analysis reveals a close phylogenetic relationship among Phaseolus vulgaris symbionts
More LessThe GenBank accession numbers for the sequences reported in this paper are AF217261 through AF217272 for nodC and AF218126, AF275670 and AF275671 for nifH.
The nodC and nifH genes were characterized in a collection of 83 rhizobial strains which represented 23 recognized species distributed in the genera Rhizobium, Sinorhizobium, Mesorhizobium and Bradyrhizobium, as well as unclassified rhizobia from various host legumes. Conserved primers were designed from available nucleotide sequences and were able to amplify nodC and nifH fragments of about 930 bp and 780 bp, respectively, from most of the strains investigated. RFLP analysis of the PCR products resulted in a classification of these rhizobia which was in general well-correlated with their known host range and independent of their taxonomic status. The nodC and nifH fragments were sequenced for representative strains belonging to different genera and species, most of which originated from Phaseolus vulgaris nodules. Phylogenetic trees were constructed and revealed close relationships among symbiotic genes of the Phaseolus symbionts, irrespective of their 16S-rDNA-based classification. The nodC and nifH phylogenies were generally similar, but cases of incongruence were detected, suggesting that genetic rearrangements have occurred in the course of evolution. The results support the view that lateral genetic transfer across rhizobial species and, in some instances, across Rhizobium and Sinorhizobium genera plays a role in diversification and in structuring the natural populations of rhizobia.
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Distribution of repC plasmid-replication sequences among plasmids and isolates of Rhizobium leguminosarum bv. viciae from field populations
More LessThe distribution of four classes of related plasmid replication genes (repC) within three field populations of Rhizobium leguminosarum in France, Germany and the UK was investigated using RFLP, PCR-RFLP and plasmid profile analysis. The results suggest that the four repC classes are compatible: when two or more different repC sequences are present in a strain they are usually associated with different plasmids. Furthermore, classical incompatibility studies in which a Tn5-labelled plasmid with a group IV repC sequence was transferred into field isolates by conjugation demonstrated that group IV sequences are incompatible with each other, but compatible with the other repC groups. This supports the idea that the different repC groups represent different incompatibility groups. The same field isolates were also screened for chromosomal (plac12) and symbiotic gene (nodD-F region) variation. Comparison of these and the plasmid data suggest that plasmid transfer does occur within field populations of R. leguminosarum but that certain plasmid-chromosome combinations are favoured.
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Transcriptional activity of the symbiotic plasmid of Rhizobium etli is affected by different environmental conditions
More LessGlobal patterns of transcriptional activity of the symbiotic plasmid (pSym) of Rhizobium etli were studied under a variety of environmental conditions, including some relevant to the symbiotic process. 32P-labelled single-stranded complementary DNAs synthesized from total RNA were used as hybridization probes against an ordered collection of cosmid clones that covered the whole pSym. Our results showed that, under aerobic conditions, discrete regions of the pSym are differentially transcribed depending on the carbon and nitrogen sources employed. In general, poor carbon or nitrogen sources allowed greater expression than rich ones. Time-course experiments with the nod gene inducer genistein led us to the identification of new regions responsive to this flavonoid. Widespread transcription was observed during microaerobiosis, but not under aerobic conditions, indicating that oxygen concentration is a major effector of transcriptional activity in the pSym. This response was reduced, but not suppressed, in a nifA mutant, indicating the location of regions whose transcription may depend on other oxygen-sensitive regulators. During symbiosis, almost the entire pSym was actively transcribed and the transcription pattern was similar to that observed during microaerobiosis. The experimental approach described allowed the identification and localization of specific regions in the pSym whose expression depends on defined environmental stimuli.
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Nitrogen limitation of chemostat-grown Rhizobium etli elicits higher infection-thread formation in Phaseolus vulgaris
More LessThe symbiotic association between rhizobia and legume roots is a complex process involving many steps. An infection thread is a tubular structure of host origin formed during the infection of legume roots by rhizobia. Previous studies with batch cultures have reported that optimal attachment of rhizobia to root hairs coincides with nutrient limitation. In this study, the ability of chemostat-grown, nutrient-limited Rhizobium etli cells to form infection threads with its symbiotic partner Phaseolus vulgaris was investigated. Rhizobia were grown in a chemostat in synthetic media under C- or N-limiting conditions. Infection-thread formation was examined after inoculation of seedlings with a rhizobial cell suspension from each treatment. The number of infection threads was estimated by light microscopy after staining root sections with o-toluidine. Exopolysaccharide (EPS) production was also measured, and the cellular content and electrophoretic pattern of lipopolysaccharide (LPS) determined semiquantitatively. N-limited cells showed a markedly higher infectivity (measured as infection-thread formation) than C-limited cells. With one of the two bean cultivars used, the number of infection threads produced by N-limited cells was higher than that produced by exponentially growing cells in batch cultures. The higher infectivity of N-limited cells was correlated with higher EPS production. Electrophoretic analysis of LPS showed that C- and N-limited cells shared a common profile but the relative concentration of short LPS forms differed.
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A metabolic study of Buchnera, the intracellular bacterial symbionts of the pea aphid Acyrthosiphon pisum
More LessCells of the bacterium Buchnera were isolated from embryos of the pea aphid Acyrthosiphon pisum, with an intact perisymbiont membrane (the insect membrane which surrounds each bacterial cell inside the aphid). The bacterial preparations respired aerobically, consuming oxygen at an average rate of 24 nmol (mg protein)−1 min−1. The bacteria took up a range of carboxylic acids and the amino acids glutamate and aspartate from an external concentration of 0·5 mm at rates of 1–10 nmol (mg protein)−1 h−1; glucose was taken up at 0·17 nmol (mg protein)−1 h−1. Glutamate uptake was proportional to its external concentration, at all concentrations tested between 15 μm and 10 mm. Saturable systems for the uptake of succinate and aspartate were identified. The kinetic constants were: K m 0·79 mm, V max 12·6 nmol (mg protein)−1 min−1 for succinate; and K m 0·22 mm, V max 3·3 nmol (mg protein)−1 min−1 for aspartate. Succinate uptake was not inhibited by the uncoupler CCCP and was markedly stimulated by ATP, suggesting that its transport is not linked to a proton-motive force but is dependent on an energized membrane and possibly mediated by a co-transport system involving another ion.
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Phage Induction of Lysogenic Rhizobium leguminosarum biovar trifolii in both the Free-living and the Symbiotic Form
More LessA lysogenic strain, Rhizobium leguminosarum biovar trifolii UK-1: φU, was isolated from a wild white clover nodule. It was symbiotically effective on white clover. A lysogenic phage (U-mole) was induced from this strain by treatment with either UV irradiation or mitomycin C. Phage U-mole had an icosahedral head (40 nm wide), a short tail (9 nm long) and tail fibres (15 nm long). Phage U-mole was induced with mitomycin C both from bacterial cells in infection threads and from bacteroids in nodules. Southern hybridization using EcoRI fragments of phage U-mole as probe indicated that phage U-mole DNA was integrated into the chromosome of strain UK-1: øU at a site involving the 60 kb EcoRI fragment of phage U-mole. Phage U-mole also lysogenized the wild-type strain R. leguminosarum biovar trifolii 4S. The resulting lysogenized strain, 4S:øU, had lost its 315 kb Sym plasmid and its nodulation ability.
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Symbiotic Plasmid Rearrangement in a Hyper-recombinant Mutant of Rhizobium leguminosarum biovar phaseoli
More LessThe symbiotic plasmid of Rhizobium leguminosarum biovar phaseoli strain CFN23 has been previously shown to undergo a series of genetic rearrangements that modify the bacterial symbiotic phenotype. A hyper-recombinant derivative of strain CFN23 was isolated. This derivative (strain CFN2300) has a similar phenotype to that reported for Escherichia coli mutants defective in DNA metabolism. The frequency of CFN23 symbiotic plasmid deletion is increased in the CFN2300 background, suggesting that homologous genetic recombination is involved in the generation of this genetic rearrangement.
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Chemotaxis of Rhizobium leguminosarum biovar phaseoli towards Flavonoid Inducers of the Symbiotic Nodulation Genes
Chemotaxis of Rhizobium leguminosarum biovar phaseoli RP8002 towards a range of carbohydrates, phenolic compounds and flavonoids was assayed. Xylose (peak response 10−4 M), sucrose (peak response 10−6 M) and raffinose (peak response 10−5 M) were strong chemoattractants amongst the carbohydrates, whilst glucose, fructose, galactose and maltose produced little or no detectable response. Of the monocyclic phenolic compounds, vanillyl alcohol, p-hydroxybenzoic acid (both peak responses 10−6 M) and 3,4-dihydroxybenzoic acid (peak response 10−4 M) all evoked strong chemotactic responses. Amongst the nod-inducing flavonoids, apigenin and luteolin were both strong chemoattractants (peaks at 10−5 M) while naringenin produced a very low response. Competition experiments suggest that apigenin and luteolin are recognized by a common receptor, but that there exists a separate receptor for luteolin alone. The inhibitors of nod-induction, umbelliferone and acetosyringone, both produced strong chemotactic responses, with peaks at 10−3 M and 10−2 M respectively. This evidence is indicative of a role for chemotaxis towards nod-inducing flavonoids in the initiation of root nodule formation by rhizobia, and also suggests that chemotaxis may influence the host range of the interaction.
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Symbiotic Phenotypes of Auxotrophic Mutants of Rhizobium meliloti104A14
More LessAuxotrophic mutants of Rhizobium meliloti 104A14 were isolated using nitrous acid mutagenesis followed by penicillin enrichment. Mutants in ornithine transcarbamylase, argininosuccinate synthetase or serine-glycine biosynthesis formed nitrogen-fixing (Fix+) nodules on the roots of alfalfa (Medicago sativa). Mutants with defects in ornithine, pyrimidine, purine, asparagine, leucine, methionine or tyrosine biosynthesis, in one-carbon metabolism or in carbamoylphosphate synthetase formed nodules but these nodules were unable to fix nitrogen. Prototrophic revertants were always Fix+. Plasmids that would complement many of these auxotrophs were isolated by transduction with a P2 cosmid gene bank of R. meliloti 104A14. These plasmids were then introduced into mutants of the same and different classes and the growth and symbiotic phenotypes of the new strains were determined. In all cases, complementation of the nutritional defect restored symbiotic nitrogen fixation.
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Plasmids and Symbiotic Effectiveness of Representative Phage Types from Two Indigenous Populations of Rhizobium meliloti
More LessSUMMARY: Phage types representative of the population of indigenous Rhizobium meliloti at each of two sites were evaluated for plasmid content by agarose gel electrophoresis and for symbiotic effectiveness with Medicago sativa cv. Saranac. Relative to four strains used commercially, 55 and 65 phage types representing these populations showed a high average level of symbiotic effectiveness; only a single type from one site was relatively ineffective in symbiosis. On the basis of plasmid number and molecular mass, 160 isolates comprising 45 and 48 types from both sites were placed in 22 different groups with 17 and 13 groups from the respective sites. The number of plasmids varied between one and five per isolate with molecular masses ranging from 5 MDa to considerably greater than 267 MDa. Only five isolates lacked a plasmid with mobility in agarose gels corresponding to that of a reference megaplasmid but instead showed a band of lesser mobility and therefore greater molecular mass. Phage types, which were divided into plasmid groups solely on the basis of differences between isolates from each site, may reflect adaptation of R. meliloti to their respective sites. Differences between isolates within certain phage types due to the presence or absence of a single plasmid may have resulted from genetic interchange between indigenous R. meliloti. There was no significant correlation between plasmid number or mass and symbiotic effectiveness or phage sensitivity of the phage types from either site.
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Ammonia Transport in Free-living and Symbiotic Rhizobium sp. ANU289
More LessSummary: [14C]Methylamine uptake by free-living Rhizobium sp. ANU289 had Michaelis–Menten kinetics (apparent K m 6.6 μm). Uptake was competitively inhibited by ammonia (K i 0.4 μm) and was dependent on an energized membrane. Uptake by bacteria had an optimum at pH 7.0. Methylamine uptake by bacteroids from siratro root nodules was much slower than that by free-living bacteria at pH 7.0 but increased exponentially with the pH of the medium. Uptake by bacteroids did not show saturation kinetics and was insensitive to the presence of ammonia or uncouplers. These results suggest that free-living bacteria (grown under conditions where ammonia is limiting) have an active transport mechanism for the uptake of ammonium ions; this carrier is not operative in the symbiotic state, where passive diffusion of ammonia occurs. In the free-living state, the ammonium carrier is under genetic control, being repressed by growth on high concentrations of ammonia. Derepression occurs under conditions of nitrogen starvation.
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Effect of Transfer of Symbiotic Plasmids and of Hydrogenase Genes (hup) on Symbiotic Efficiency of Rhizobium leguminosarum Strains
More LessSummary: Plasmid-encoded symbiotic determinants from the Rhizobium leguminosarum strain MA1 (817) with uptake hydrogenase activity (Hup+) and from the Hup− strain MC1 (18a) were mobilized by recombination with the self-transmissible plasmid pVW5JI. The symbiotic determinants were transferred by conjugation from strain MA1 to strain MC1 and to a derivative of MC1 without the symbiotic plasmid, and vice versa, thus constructing four types of transconjugants. The determinants for a total recycling hydrogenase in strain MA1 were found to be encoded on the symbiotic plasmid.
Strain MC1 fixed 60% more N2 in pea root nodules, determined as mg nitrogen per plant, than strain MA1. This difference was not increased in the MC1 derivative that obtained hydrogenase activity. Plants inoculated with a derivative of strain MA1, however, where the symbiotic plasmid was replaced by that of strain MC1 had a high percentage nitrogen content. It was concluded that the symbiotic plasmid and the genetic background were more important for plant nitrogen accumulation than uptake hydrogenase.
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Sugar Metabolism and the Symbiotic Properties of Carbohydrate Mutants of Rhizobium leguminosarum
More LessRhizobium leguminosarum metabolizes sugars via the Entner-Doudoroff and pentose phosphate pathways but does not have a functional Embden-Meyerhof pathway. Although some sugar catabolizing enzymes are constitutive, activities of the ‘Entner-Doudoroff’ enzymes vary with the carbon source. Bacteroids have complete pathways for sugar catabolism even though the specific activities of some enzymes, e.g., glucokinase, are lower than in free-living cells. Tn5-induced mutants lacking glucokinase, fructokinase and pyruvate dehydrogenase have been isolated. Although these mutants are unable to utilize sugars, they all nodulate peas and fix N2. The capacity to utilize particular C6 and C12 sugars is apparently not essential for bacteroid development or the establishment of effective N2 fixation.
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Contribution of the Symbiotic Plasmid to the Competitiveness of Rhizobium leguminosarum
More LessFour different symbiotic plasmids from Rhizobium leguminosarum were introduced into three different recipient strains that lacked plasmid-linked symbiotic determinants. The twelve synthetic strains so constructed were each tested for competitiveness against a standard reference strain. The recipient strain and the introduced symbiotic plasmid contributed about equally to competitiveness in forming root nodules on pea plants: there was also significant interaction between strain and plasmid, although this was much less important than the main effects. Competitiveness for growth on the legume root surface (the rhizosphere) was attributable entirely to the recipient strain; the introduced plasmid had no significant effect.
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Antibiotic Activity of Xenorhabdus spp., Bacteria Symbiotically Associated with Insect Pathogenic Nematodes of the Families Heterorhabditidae and Steinernematidae
More LessA wide range of micro-organisms, including yeasts, was found to be inhibited by the primary form of Xenorhabdus spp., but not by the secondary form. Only one Xenorhabdus strain, the symbiont of Neoaplectana glaseri, did not inhibit any of the micro-organisms tested; it is suggested that this strain may not have been isolated in the primary form. Gram-positive bacteria were sensitive to all active isolates of Xenorhabdus; each of the yeasts and almost all of the Gram-negative bacteria were sensitive to some but not all Xenorhabdus isolates. Each Xenorhabdus isolate was sensitive to some other Xenorhabdus isolates. The antibiotic activity of X. nematophilus was unaffected by autoclaving but was lost after dialysis. Anaerobically incubated Xenorhabdus spp. did not exhibit antibiotic activity.
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