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1. Entcheva  P, Phillips  DA, Streit  WR,     ( 2002 )

Functional analysis of Sinorhizobium meliloti genes involved in biotin synthesis and transport.

Applied and environmental microbiology 68 (6)
PMID : 12039741  :   DOI  :   10.1128/aem.68.6.2843-2848.2002     PMC  :   PMC123963    
Abstract >>
External biotin greatly stimulates bacterial growth and alfalfa root colonization by Sinorhizobium meliloti strain 1021. Several genes involved in responses to plant-derived biotin have been identified in this bacterium, but no genes required for biotin transport are known, and not all loci required for biotin synthesis have been assigned. Searches of the S. meliloti genome database in combination with complementation tests of Escherichia coli biotin auxotrophs indicate that biotin synthesis probably is limited in S. meliloti 1021 by the poor functioning or complete absence of several key genes. Although several open reading frames with significant similarities to genes required for synthesis of biotin in gram-positive and gram-negative bacteria were found, only bioB, bioF, and bioH were demonstrably functional in complementation tests with known E. coli mutants. No sequence or complementation evidence was found for bioA, bioC, bioD, or bioZ. In contrast to other microorganisms, the S. meliloti bioB and bioF genes are not localized in a biotin synthesis operon, but bioB is cotranscribed with two genes coding for ABC transporter-like proteins, designated here bioM and bioN. Mutations in bioM and bioN eliminated growth on alfalfa roots and reduced bacterial capacity to maintain normal intracellular levels of biotin. Taken together, these data suggest that S. meliloti normally grows on exogenous biotin using bioM and bioN to conserve biotin assimilated from external sources.
KeywordMeSH Terms
Transaminases
2. Sullivan  JT, Trzebiatowski  JR, Cruickshank  RW, Gouzy  J, Brown  SD, Elliot  RM, Fleetwood  DJ, McCallum  NG, Rossbach  U, Stuart  GS, Weaver  JE, Webby  RJ, De Bruijn  FJ, Ronson  CW,     ( 2002 )

Comparative sequence analysis of the symbiosis island of Mesorhizobium loti strain R7A.

Journal of bacteriology 184 (11)
PMID : 12003951  :   DOI  :   10.1128/jb.184.11.3086-3095.2002     PMC  :   PMC135072    
Abstract >>
The Mesorhizobium loti strain R7A symbiosis island is a 502-kb chromosomally integrated element which transfers to nonsymbiotic mesorhizobia in the environment, converting them to Lotus symbionts. It integrates into a phenylalanine tRNA gene in a process mediated by a P4-type integrase encoded at the left end of the element. We have determined the nucleotide sequence of the island and compared its deduced genetic complement with that reported for the 611-kb putative symbiosis island of M. loti strain MAFF303099. The two islands share 248 kb of DNA, with multiple deletions and insertions of up to 168 kb interrupting highly conserved colinear DNA regions in the two strains. The shared DNA regions contain all the genes likely to be required for Nod factor synthesis, nitrogen fixation, and island transfer. Transfer genes include a trb operon and a cluster of potential tra genes which are also present on the strain MAFF303099 plasmid pMLb. The island lacks plasmid replication genes, suggesting that it is a site-specific conjugative transposon. The R7A island encodes a type IV secretion system with strong similarity to the vir pilus from Agrobacterium tumefaciens that is deleted from MAFF303099, which in turn encodes a type III secretion system not found on the R7A island. The 414 genes on the R7A island also include putative regulatory genes, transport genes, and an array of metabolic genes. Most of the unique hypothetical genes on the R7A island are strain-specific and clustered, suggesting that they may represent other acquired genetic elements rather than symbiotically relevant DNA.
KeywordMeSH Terms
Genes, Bacterial
Symbiosis
3. Lepek  VC, D'Antuono  AL, Tomatis  PE, Ugalde  JE, Giambiagi  S, Ugalde  RA,     ( 2002 )

Analysis of Mesorhizobium loti glycogen operon: effect of phosphoglucomutase (pgm) and glycogen synthase (g/gA) null mutants on nodulation of Lotus tenuis.

Molecular plant-microbe interactions : MPMI 15 (4)
PMID : 12026175  :   DOI  :   10.1094/MPMI.2002.15.4.368    
Abstract >>
The phosphoglucomutase (pgm) gene codes for a key enzyme required for the formation of UDP-glucose and ADP-glucose, the sugar donors for the biosynthesis of glucose containing polysaccharides. A Mesorhizobium loti pgm null mutant obtained in this study contains an altered form of lipopolysaccharide (LPS), lacks exopolysaccharide (EPS), beta cyclic glucan, and glycogen and is unable to nodulate Lotus tenuis. The nonnodulating phenotype of the pgm mutant was not due to the absence of glycogen, since a glycogen synthase (glgA) null mutant effectively nodulates this legume. In M. loti, pgm is part of the glycogen metabolism gene cluster formed by GlgP (glycogen phosphorylase), glgB (glycogen branching), glgC (ADP-glucose pyrophosphorylase), glgA, pgm, and glgX (glycogen debranching). The genes are transcribed as a single transcript from glgP to at least pgm under the control of a strong promoter (promoter I) upstream of glgP. An alternative promoter (promoter II), mapping in a 154-bp DNA fragment spanning 85 bp upstream of the glgA start codon and the first 69 bp of the glgA coding region, controls the expression of glgA and pgm, independently of the rest of the upstream genes. Primer extension experiments showed that transcription starts 19 bp upstream of the glgA start codon.
KeywordMeSH Terms
4. Gaunt  MW, Turner  SL, Rigottier-Gois  L, Lloyd-Macgilp  SA, Young  JP,     ( 2001 )

Phylogenies of atpD and recA support the small subunit rRNA-based classification of rhizobia.

International journal of systematic and evolutionary microbiology 51 (Pt 6)
PMID : 11760945  :   DOI  :   10.1099/00207713-51-6-2037    
Abstract >>
The current classification of the rhizobia (root-nodule symbionts) assigns them to six genera. It is strongly influenced by the small subunit (16S, SSU) rRNA molecular phylogeny, but such single-gene phylogenies may not reflect the evolution of the genome as a whole. To test this, parts of the atpD and recA genes have been sequenced for 25 type strains within the alpha-Proteobacteria, representing species in Rhizobium, Sinorhizobium, Mesorhizobium, Bradyrhizobium, Azorhizobium, Agrobacterium, Phyllobacterium, Mycoplana and Brevundimonas. The current genera Sinorhizobium and Mesorhizobium are well supported by these genes, each forming a distinct phylogenetic clade with unequivocal bootstrap support. There is good support for a Rhizobium clade that includes Agrobacterium tumefaciens, and the very close relationship between Agrobacterium rhizogenes and Rhizobium tropici is confirmed. There is evidence for recombination within the genera Mesorhizobium and Sinorhizobium, but the congruence of the phylogenies at higher levels indicates that the genera are genetically isolated. rRNA provides a reliable distinction between genera, but genetic relationships within a genus may be disturbed by recombination.
KeywordMeSH Terms
Phylogeny
5. López-Lara  IM, Kafetzopoulos  D, Spaink  HP, Thomas-Oates  JE,     ( 2001 )

Rhizobial NodL O-acetyl transferase and NodS N-methyl transferase functionally interfere in production of modified Nod factors.

Journal of bacteriology 183 (11)
PMID : 11344149  :   DOI  :   10.1128/JB.183.11.3408-3416.2001     PMC  :   PMC99639    
Abstract >>
The products of the rhizobial nodulation genes are involved in the biosynthesis of lipochitin oligosaccharides (LCOs), which are host-specific signal molecules required for nodule formation. The presence of an O-acetyl group on C-6 of the nonreducing N-acetylglucosamine residue of LCOs is due to the enzymatic activity of NodL. Here we show that transfer of the nodL gene into four rhizobial species that all normally produce LCOs that are not modified on C-6 of the nonreducing terminal residue results in production of LCOs, the majority of which have an acetyl residue substituted on C-6. Surprisingly, in transconjugant strains of Mesorhizobium loti, Rhizobium etli, and Rhizobium tropici carrying nodL, such acetylation of LCOs prevents the endogenous nodS-dependent transfer of the N-methyl group that is found as a substituent of the acylated nitrogen atom. To study this interference between nodL and nodS, we have cloned the nodS gene of M. loti and used its product in in vitro experiments in combination with purified NodL protein. It has previously been shown that a chitooligosaccharide N deacetylated on the nonreducing terminus (the so-called NodBC metabolite) is the preferred substrate for NodS as well as for NodL. Here we show that the NodBC metabolite, acetylated by NodL, is not used by the NodS protein as a substrate while the NodL protein can acetylate the NodBC metabolite that has been methylated by NodS.
KeywordMeSH Terms
6. Sullivan  JT, Brown  SD, Yocum  RR, Ronson  CW,     ( 2001 )

The bio operon on the acquired symbiosis island of Mesorhizobium sp. strain R7A includes a novel gene involved in pimeloyl-CoA synthesis.

Microbiology (Reading, England) 147 (Pt 5)
PMID : 11320134  :   DOI  :   10.1099/00221287-147-5-1315    
Abstract >>
The symbiosis island of Mesorhizobium sp. strain R7A is a 500 kb chromosomal genetic element that upon transfer converts nonsymbiotic mesorhizobia to symbionts able to nodulate and fix nitrogen with Lotus corniculatus. Four genomic species of nonsymbiotic mesorhizobia have been isolated. All were auxotrophic for thiamin and biotin and three were auxotrophic for nicotinate, whereas derivatives of the strains containing the symbiosis island were prototrophic for all three vitamins. In this work, a 13.2 kb region of the island that converts the nonsymbionts to nicotinate and biotin prototrophy was characterized. The region contained orthologues of the Escherichia coli bioBFD and A genes arranged in an operon with a novel gene, bioZ, a nadABC operon, the nitrogen-fixation regulatory gene nifA, and a homologue of the pantothenate biosynthesis gene panD. The bioZ gene product was similar to beta-ketoacyl-acyl carrier protein synthase III (FabH). bioZ::Tn5 mutants grew poorly in the absence of biotin and the bioZ gene complemented an E. coli bioH mutant, suggesting that its product is involved in the synthesis of pimeloyl-COA: The bio operon was not required for symbiosis, as only mutants in the nifA gene were impaired in symbiosis, and a bioA::Tn5 mutant was not impaired in rhizosphere colonization. The rationale for the vitamin biosynthetic loci being located on an acquired genetic element that is absent from nonsymbiotic mesorhizobia remains to be determined.
KeywordMeSH Terms
Genes, Bacterial
Operon
Symbiosis
7. Sy  A, Giraud  E, Jourand  P, Garcia  N, Willems  A, de Lajudie  P, Prin  Y, Neyra  M, Gillis  M, Boivin-Masson  C, Dreyfus  B,     ( 2001 )

Methylotrophic Methylobacterium bacteria nodulate and fix nitrogen in symbiosis with legumes.

Journal of bacteriology 183 (1)
PMID : 11114919  :   DOI  :   10.1128/JB.183.1.214-220.2001     PMC  :   PMC94868    
Abstract >>
Rhizobia described so far belong to three distinct phylogenetic branches within the alpha-2 subclass of Proteobacteria. Here we report the discovery of a fourth rhizobial branch involving bacteria of the Methylobacterium genus. Rhizobia isolated from Crotalaria legumes were assigned to a new species, "Methylobacterium nodulans," within the Methylobacterium genus on the basis of 16S ribosomal DNA analyses. We demonstrated that these rhizobia facultatively grow on methanol, which is a characteristic of Methylobacterium spp. but a unique feature among rhizobia. Genes encoding two key enzymes of methylotrophy and nodulation, the mxaF gene, encoding the alpha subunit of the methanol dehydrogenase, and the nodA gene, encoding an acyltransferase involved in Nod factor biosynthesis, were sequenced for the type strain, ORS2060. Plant tests and nodA amplification assays showed that "M. nodulans" is the only nodulating Methylobacterium sp. identified so far. Phylogenetic sequence analysis showed that "M. nodulans" NodA is closely related to Bradyrhizobium NodA, suggesting that this gene was acquired by horizontal gene transfer.
KeywordMeSH Terms
Plants, Medicinal
Symbiosis
8. Turner  SL,     ( 2000 )

The glutamine synthetases of rhizobia: phylogenetics and evolutionary implications.

Molecular biology and evolution 17 (2)
PMID : 10677854  :   DOI  :   10.1093/oxfordjournals.molbev.a026311    
Abstract >>
Glutamine synthetase exists in at least two related forms, GSI and GSII, the sequences of which have been used in evolutionary molecular clock studies. GSI has so far been found exclusively in bacteria, and GSII has been found predominantly in eukaryotes. To date, only a minority of bacteria, including rhizobia, have been shown to express both forms of GS. The sequences of equivalent internal fragments of the GSI and GSII genes for the type strains of 16 species of rhizobia have been determined and analyzed. The GSI and GSII data sets do not produce congruent phylogenies with either neighbor-joining or maximum-likelihood analyses. The GSI phylogeny is broadly congruent with the 16S rDNA phylogeny for the same bacteria; the GSII phylogeny is not. There are three striking rearrangements in the GSII phylograms, all of which might be explained by horizontal gene transfer to Bradyrhizobium (probably from Mesorhizobium), to Rhizobium galegae (from Rhizobium), and to Mesorhizobium huakuii (perhaps from Rhizobium). There is also evidence suggesting intrageneric DNA transfer within Mesorhizobium. Meta-analysis of both GS genes from the different genera of rhizobia and other reference organisms suggests that the divergence times of the different rhizobium genera predate the existence of legumes, their host plants.
KeywordMeSH Terms
Evolution, Molecular
Phylogeny
9. Martens  M, Delaere  M, Coopman  R, De Vos  P, Gillis  M, Willems  A,     ( 2007 )

Multilocus sequence analysis of Ensifer and related taxa.

International journal of systematic and evolutionary microbiology 57 (Pt 3)
PMID : 17329774  :   DOI  :   10.1099/ijs.0.64344-0    
Abstract >>
Multilocus sequence analysis (MLSA) was performed on representatives of Ensifer (including species previously assigned to the genus Sinorhizobium) and related taxa. Neighbour-joining (NJ), maximum-parsimony (MP) and maximum-likelihood (ML) phylogenies of dnaK, gltA, glnA, recA, thrC and 16S rRNA genes were compared. The data confirm that the potential for discrimination of Ensifer species is greater using MLSA of housekeeping genes than 16S rRNA genes. In incongruence-length difference tests, the 16S rRNA gene was found to be significantly incongruent with the other genes, indicating that this gene should not be used as a single indicator of relatedness in this group. Significant congruence was detected for dnaK, glnA and thrC. Analyses of concatenated sequences of dnaK, glnA and thrC genes yielded very similar NJ, MP and ML trees, with high bootstrap support. In addition, analysis of a concatenation of all six genes essentially produced the same result, levelling out potentially conflicting phylogenetic signals. This new evidence supports the proposal to unite Ensifer and Sinorhizobium in a single genus. Support for an alternative solution preserving the two genera is less strong. In view of the opinions expressed by the Judicial Commission, the name of the genus should be Ensifer, as proposed by Young [Young, J. M. (2003). Int J Syst Evol Microbiol 53, 2107-2110]. Data obtained previously and these new data indicate that Ensifer adhaerens and 'Sinorhizobium morelense' are not heterotypic synonyms, but represent separate species. However, transfer to the genus Ensifer is not possible at present because the species name is the subject of a pending Request for an Opinion, which would affect whether a novel species in the genus Ensifer or a new combination based on a basonym would be created.
KeywordMeSH Terms
10. Yuan  B, Yoshikane  Y, Yokochi  N, Ohnishi  K, Yagi  T,     ( 2004 )

The nitrogen-fixing symbiotic bacterium Mesorhizobium loti has and expresses the gene encoding pyridoxine 4-oxidase involved in the degradation of vitamin B6.

FEMS microbiology letters 234 (2)
PMID : 15135526  :   DOI  :   10.1016/j.femsle.2004.03.031    
Abstract >>
The gene product of mll6785 of a nitrogen-fixing symbiotic bacterium Mesorhizobium loti MAFF303099 was identified as pyridoxine 4-oxidase, the first enzyme in the vitamin B6-degradation pathway. The gene was cloned and ligated into pET-21a+. Escherichia coli BL21(DE3) was co-transformed with the constructed plasmid plus pKY206 containing groESL genes encoding chaperonins. The overexpressed protein was purified to homogeneity by the ammonium sulfate fractionation and three chromatography steps. The enzymatic properties of the purified protein, such as K(m) values for pyridoxine (213+/-19 microM) and oxygen (78+/-10 microM), were compared to those of pyridoxine 4-oxidase from two bacteria with known vitamin B6-degradation pathway. M. loti grown in a Rhizobium medium showed the enzyme activity. The results suggest that M. loti also contains the degradation pathway of vitamin B6.
KeywordMeSH Terms
11. Stepkowski  T, Czapli?ska  M, Miedzinska  K, Moulin  L,     ( 2003 )

The variable part of the dnaK gene as an alternative marker for phylogenetic studies of rhizobia and related alpha Proteobacteria.

Systematic and applied microbiology 26 (4)
PMID : 14666974  :   DOI  :   10.1078/072320203770865765    
Abstract >>
DnaK is the 70 kDa chaperone that prevents protein aggregation and supports the refolding of damaged proteins. Due to sequence conservation and its ubiquity this chaperone has been widely used in phylogenetic studies. In this study, we applied the less conserved part that encodes the so-called alpha-subdomain of the substrate-binding domain of DnaK for phylogenetic analysis of rhizobia and related non-symbiotic alpha-Proteobacteria. A single 330 bp DNA fragment was routinely amplified from DNA templates isolated from the species of the genera, Azorhizobium, Bradyrhizobium, Mesorhizobium, Rhizobium and Sinorhizobium, but also from some non-symbiotic alpha Proteobacteria such as Blastochloris, Chelatobacter and Chelatococcus. Phylogenetic analyses revealed high congruence between dnaK sequences and 16S rDNA trees, but they were not identical. In contrast, the partition homogeneity tests revealed that dnaK sequence data could be combined with other housekeeping genes such as recA, atpD or glnA. The dnaK trees exhibited good resolution in the cases of the genera Mesorhizobium, Sinorhizobium and Rhizobium, even better than usually shown by 16S rDNA phylogeny. The dnaK phylogeny supported the close phylogenetic relationship of Rhizobium galegae and Agrobacterium tumefaciens (R. radiobacter) C58, which together formed a separate branch within the fast-growing rhizobia, albeit closer to the genus Sinorhizobium. The Rhizobium and Sinorhizobium genera carried an insertion composed of two amino acids, which additionally supported the phylogenetic affinity of these two genera, as well as their distinctness from the Mesorhizobium genus. Consistently with the phylogeny shown by 16S-23S rDNA intergenic region sequences, the dnaK trees divided the genus Bradyrhizobium into three main lineages, corresponding to B. japonicum, B. elkanii, and photosynthetic Bradyrhizobium strains that infect Aeschynomene plants. Our results suggest that the 330 bp dnaK sequences could be used as an additional taxonomic marker for rhizobia and related species (alternatively to the 16S rRNA gene phylogeny).
KeywordMeSH Terms
Escherichia coli Proteins
Phylogeny
Soil Microbiology
12. Ampomah  OY, Huss-Danell  K,     ( 2011 )

Genetic diversity of root nodule bacteria nodulating Lotus corniculatus and Anthyllis vulneraria in Sweden.

Systematic and applied microbiology 34 (4)
PMID : 21497473  :   DOI  :   10.1016/j.syapm.2011.01.006    
Abstract >>
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.
KeywordMeSH Terms
Biodiversity
Genetic Variation
13. Díaz  R, Vargas-Lagunas  C, Villalobos  MA, Peralta  H, Mora  Y, Encarnación  S, Girard  L, Mora  J,     ( 2011 )

argC Orthologs from Rhizobiales show diverse profiles of transcriptional efficiency and functionality in Sinorhizobium meliloti.

Journal of bacteriology 193 (2)
PMID : 21075924  :   DOI  :   10.1128/JB.01010-10     PMC  :   PMC3019832    
Abstract >>
Several factors can influence ortholog replacement between closely related species. We evaluated the transcriptional expression and metabolic performance of ortholog substitution complementing a Sinorhizobium meliloti argC mutant with argC from Rhizobiales (Agrobacterium tumefaciens, Rhizobium etli, and Mesorhizobium loti). The argC gene is necessary for the synthesis of arginine, an amino acid that is central to protein and cellular metabolism. Strains were obtained carrying plasmids with argC orthologs expressed under the speB and argC (S. meliloti) and lac (Escherichia coli) promoters. Complementation analysis was assessed by growth, transcriptional activity, enzymatic activity, mRNA levels, specific detection of ArgC proteomic protein, and translational efficiency. The argC orthologs performed differently in each complementation, reflecting the diverse factors influencing gene expression and the ability of the ortholog product to function in a foreign metabolic background. Optimal complementation was directly related to sequence similarity with S. meliloti, and was inversely related to species signature, with M. loti argC showing the poorest performance, followed by R. etli and A. tumefaciens. Different copy numbers of genes and amounts of mRNA and protein were produced, even with genes transcribed from the same promoter, indicating that coding sequences play a role in the transcription and translation processes. These results provide relevant information for further genomic analyses and suggest that orthologous gene substitutions between closely related species are not completely functionally equivalent.
KeywordMeSH Terms
Gene Expression Regulation, Bacterial
14. Shiraishi  A, Matsushita  N, Hougetsu  T,     ( 2010 )

Nodulation in black locust by the Gammaproteobacteria Pseudomonas sp. and the Betaproteobacteria Burkholderia sp.

Systematic and applied microbiology 33 (5)
PMID : 20542651  :   DOI  :   10.1016/j.syapm.2010.04.005    
Abstract >>
Nodulation abilities of bacteria in the subclasses Gammaproteobacteria and Betaproteobacteria on black locust (Robinia pseudoacacia) were tested. Pseudomonas sp., Burkholderia sp., Klebsiella sp., and Paenibacillus sp. were isolated from surface-sterilized black locust nodules, but their nodulation ability is unknown. The aims of this study were to determine if these bacteria are symbiotic. The species and genera of the strains were determined by RFLP analysis and DNA sequencing of 16S rRNA gene. Inoculation tests and histological studies revealed that Pseudomonas sp. and Burkholderia sp. formed nodules on black locust and also developed differentiated nodule tissue. Furthermore, a phylogenetic analysis of nodA and a BLASTN analysis of the nodC, nifH, and nifHD genes revealed that these symbiotic genes of Pseudomonas sp. and Burkholderia sp. have high similarities with those of rhizobial species, indicating that the strains acquired the symbiotic genes from rhizobial species in the soil. Therefore, in an actual rhizosphere, bacterial diversity of nodulating legumes may be broader than expected in the Alpha-, Beta-, and Gammaproteobacteria subclasses. The results indicate the importance of horizontal gene transfer for establishing symbiotic interactions in the rhizosphere.
KeywordMeSH Terms
Betaproteobacteria
Gammaproteobacteria
Symbiosis
15. Yokochi  N, Yoshikane  Y, Matsumoto  S, Fujisawa  M, Ohnishi  K, Yagi  T,     ( 2009 )

Gene identification and characterization of 5-formyl-3-hydroxy-2-methylpyridine 4-carboxylic acid 5-dehydrogenase, an NAD+-dependent dismutase.

Journal of biochemistry 145 (4)
PMID : 19218190  :   DOI  :   10.1093/jb/mvp007    
Abstract >>
A chromosomal gene, mlr6793, in Mesorhizobium loti was identified as the gene encoding 5-formyl-3-hydroxy-2-methylpyridine 4-carboxylic acid (FHMPC) dehydrogenase (dismutase) involved in the degradation pathway for pyridoxine (vitamin B(6)). The homogenously purified recombinant enzyme has a molecular mass of 59.1 kDa and is a homodimeric protein. FHMPC dehydrogenase catalyses practically irreversible oxidation (k(cat) = 204 s(-1)) of FHMPC (K(m) = 48.2 microM) by NAD(+) (K(m) = 34.3 microM) to 3-hydroxy-2-methyl-pyridine 4, 5-dicarboxylic acid (HMPDC), and practically irreversible reduction (k(cat) = 217 s(-1)) of FHMPC (K(m) = 24.9 microM) by NADH (K(m) = 12.4 microM) to 4-pyridoxic acid. When the enzyme reaction was started with the combination of FHMPC and NAD(+) or that of FHMPC and NADH, HMPDC and 4-pyridoxic acid were produced in an almost equimolar ratio throughout the reaction. FHMPC dehydrogenase belongs to the 3-hydroxyacyl-CoA dehydrogenase family with 31% identity with the human enzyme: it has probable catalytic diad residues, i.e. His137 and Glu149. The H137L mutant enzyme showed no measurable activity. The E149Q one was stable in contrast to the corresponding human 3-hydroxyacyl-CoA dehydrogenase mutant, and showed unique pH optima depending on the co-substrates used for the reaction.
KeywordMeSH Terms
Genes, Bacterial
16. Yang  M, Sun  K, Zhou  L, Yang  R, Zhong  Z, Zhu  J,     ( 2009 )

Functional analysis of three AHL autoinducer synthase genes in Mesorhizobium loti reveals the important role of quorum sensing in symbiotic nodulation.

Canadian journal of microbiology 55 (2)
PMID : 19295655  :   DOI  :   10.1139/w08-128    
Abstract >>
One of the most important signal transduction pathways in bacteria, quorum sensing, is involved in many regulatory circuits in rhizobia, especially in the control of communication between rhizobia and their plant hosts. In this study, we identified 3 autoinducer synthase genes - mrlI1, mrlI2, and mrlI3 - in Mesorhizobium loti NZP 2213. We found that MrlI1 and MrlI2 could synthesize distinct N-acyl homoserine lactone (AHL) autoinducers in rich medium cultures, and the expression of mrlI1 was shown to be growth-phase-dependent. MrlI3 did not produce any detectable AHL molecules under the culture conditions tested. To investigate whether these AHL synthases affect nodulation, we examined the nodulation of AHL-deficient mutants on their native plant host Lotus corniculatus and found that the efficiency of nodulation of bacteria with mutations of any of these 3 synthase genes was reduced, suggesting that quorum sensing systems in M. loti may play an important role in successful establishment of rhizobium-legume symbiosis.
KeywordMeSH Terms
Quorum Sensing
Symbiosis
17. Alexandre  A, Laranjo  M, Young  JP, Oliveira  S,     ( 2008 )

dnaJ is a useful phylogenetic marker for alphaproteobacteria.

International journal of systematic and evolutionary microbiology 58 (Pt 12)
PMID : 19060069  :   DOI  :   10.1099/ijs.0.2008/001636-0    
Abstract >>
In the past, bacterial phylogeny relied almost exclusively on 16S rRNA gene sequence analysis. More recently, multilocus sequence analysis has been used to infer organismal phylogenies. In this study, the dnaJ chaperone gene was investigated as a marker for phylogeny studies in alphaproteobacteria. Preliminary analysis of G+C contents and G+C3s contents (the G+C content of the synonymous third codon position) showed no clear evidence of horizontal transfer of this gene in proteobacteria. dnaJ-based phylogenies were then analysed at three taxonomic levels: the Proteobacteria, the Alphaproteobacteria and the genus Mesorhizobium. Dendrograms based on DnaJ and 16S rRNA gene sequences revealed the same topology described previously for the Proteobacteria. These results indicate that the DnaJ phylogenetic signal is able to reproduce the accepted relationships among the five classes of the Proteobacteria. At a lower taxonomic level, using 20 alphaproteobacteria, the 16S rRNA gene-based phylogeny is distinct from the one based on DnaJ sequence analysis. Although the same clusters are generated, only the topology of the DnaJ tree is consistent with broader phylogenies from recent studies based on concatenated alignments of multiple core genes. For example, the DnaJ tree shows the two clusters within the Rhizobiales as closely related, as expected, while the 16S rRNA gene-based phylogeny shows them as distantly related. In order to evaluate the phylogenetic performance of dnaJ at the genus level, a multilocus analysis based on five housekeeping genes (atpD, gapA, gyrB, recA and rplB) was performed for ten Mesorhizobium species. In contrast to the 16S rRNA gene, the DnaJ sequence analysis generated a tree similar to the multilocus dendrogram. For identification of chickpea mesorhizobium isolates, a dnaJ nucleotide sequence-based tree was used. Despite different topologies, 16S rRNA gene- and dnaJ-based trees led to the same species identification. This study suggests that the dnaJ gene is a good phylogenetic marker, particularly for the class Alphaproteobacteria, since its phylogeny is consistent with phylogenies based on multilocus approaches.
KeywordMeSH Terms
Phylogeny
18. Laranjo  M, Alexandre  A, Rivas  R, Velázquez  E, Young  JP, Oliveira  S,     ( 2008 )

Chickpea rhizobia symbiosis genes are highly conserved across multiple Mesorhizobium species.

FEMS microbiology ecology 66 (2)
PMID : 18795953  :   DOI  :   10.1111/j.1574-6941.2008.00584.x    
Abstract >>
Chickpea has been considered as a restrictive host for nodulation by rhizobia. However, recent studies have reported that several Mesorhizobium species may effectively nodulate chickpea. With the purpose of investigating the evolutionary relationships between these different species with the ability of nodulating the same host, we analysed 21 Portuguese chickpea rhizobial isolates. Symbiosis genes nifH and nodC were sequenced and used for phylogenetic studies. Symbiotic effectiveness was determined to evaluate its relationship with symbiosis genes. The comparison of 16S rRNA gene-based phylogeny with the phylogenies based on symbiosis genes revealed evidence of lateral transfer of symbiosis genes across different species. Chickpea is confirmed as a nonpromiscuous host. Although chickpea is nodulated by many different species, they share common symbiosis genes, suggesting recognition of only a few Nod factors by chickpea. Our results suggest that sequencing of nifH or nodC genes can be used for rapid detection of chickpea mesorhizobia.
KeywordMeSH Terms
Alphaproteobacteria
Gene Transfer, Horizontal
Phylogeny
19. Ge  F, Yokochi  N, Yoshikane  Y, Ohnishi  K, Yagi  T,     ( 2008 )

Gene identification and characterization of the pyridoxine degradative enzyme 4-pyridoxic acid dehydrogenase from the nitrogen-fixing symbiotic bacterium Mesorhizobium loti MAFF303099.

Journal of biochemistry 143 (5)
PMID : 18216065  :   DOI  :   10.1093/jb/mvn010    
Abstract >>
The gene encoding 4-pyridoxic acid dehydrogenase was identified as mlr6792 in a chromosome of a nitrogen-fixing symbiotic bacterium Mesorhizobium loti MAFF303099. The enzyme is the fourth enzyme in the vitamin B(6) (pyridoxine)-degradation pathway I. The recombinant enzyme with a his-tag over-expressed in Escherichia coli cells was a membrane-bound protein, and purified to homogeneity. The enzyme was a monomeric protein with a molecular weight of 59,000, and a flavoprotein containing one mole of FAD per mole of subunit. The optimum pH and temperature, and K(m) for 4-pyridoxic acid were pH 8.5 and 30 degrees C, and 29 muM, respectively. The enzyme was a glucose-methanol-choline (GMC) family protein with two signature patterns, FAD-binding residues, a putative active site histidine residue and a probable transmembrane segment.
KeywordMeSH Terms
20. Madhaiyan  M, Alex  TH, Ngoh  ST, Prithiviraj  B, Ji  L,     ( 2015 )

Leaf-residing Methylobacterium species fix nitrogen and promote biomass and seed production in Jatropha curcas.

Biotechnology for biofuels 8 (N/A)
PMID : 26697111  :   DOI  :   10.1186/s13068-015-0404-y     PMC  :   PMC4687150    
Abstract >>
Jatropha curcas L. (Jatropha) is a potential biodiesel crop that can be cultivated on marginal land because of its strong tolerance to drought and low soil nutrient content. However, seed yield remains low. To enhance the commercial viability and green index of Jatropha biofuel, a systemic and coordinated approach must be adopted to improve seed oil and biomass productivity. Here, we present our investigations on the Jatropha-associated nitrogen-fixing bacteria with an aim to understand and exploit the unique biology of this plant from the perspective of plant-microbe interactions. An analysis of 1017 endophytic bacterial isolates derived from different parts of Jatropha revealed that diazotrophs were abundant and diversely distributed into five classes belonging to �\, �], �^-Proteobacteria, Actinobacteria and Firmicutes. Methylobacterium species accounted for 69.1 % of endophytic bacterial isolates in leaves and surprisingly, 30.2 % which were able to fix nitrogen that inhabit in leaves. Among the Methylobacterium isolates, strain L2-4 was characterized in detail. Phylogenetically, strain L2-4 is closely related to M. radiotolerans and showed strong molybdenum-iron dependent acetylene reduction (AR) activity in vitro and in planta. Foliar spray of L2-4 led to successful colonization on both leaf surface and in internal tissues of systemic leaves and significantly improved plant height, leaf number, chlorophyll content and stem volume. Importantly, seed production was improved by 222.2 and 96.3 % in plants potted in sterilized and non-sterilized soil, respectively. Seed yield increase was associated with an increase in female-male flower ratio. The ability of Methylobacterium to fix nitrogen and colonize leaf tissues serves as an important trait for Jatropha. This bacteria-plant interaction may significantly contribute to Jatropha's tolerance to low soil nutrient content. Strain L2-4 opens a new possibility to improve plant's nitrogen supply from the leaves and may be exploited to significantly improve the productivity and Green Index of Jatropha biofuel.
KeywordMeSH Terms
Biofuel
Culturable endophyte
Jatropha curcas L.
Methylobacterium
Nitrogen fixation
Biofuel
Culturable endophyte
Jatropha curcas L.
Methylobacterium
Nitrogen fixation
Biofuel
Culturable endophyte
Jatropha curcas L.
Methylobacterium
Nitrogen fixation
Biofuel
Culturable endophyte
Jatropha curcas L.
Methylobacterium
Nitrogen fixation
Biofuel
Culturable endophyte
Jatropha curcas L.
Methylobacterium
Nitrogen fixation
Biofuel
Culturable endophyte
Jatropha curcas L.
Methylobacterium
Nitrogen fixation
Biofuel
Culturable endophyte
Jatropha curcas L.
Methylobacterium
Nitrogen fixation
Biofuel
Culturable endophyte
Jatropha curcas L.
Methylobacterium
Nitrogen fixation
21. Ji  Z, Yan  H, Cui  Q, Wang  E, Chen  W, Chen  W,     ( 2015 )

Genetic divergence and gene flow among Mesorhizobium strains nodulating the shrub legume Caragana.

Systematic and applied microbiology 38 (3)
PMID : 25864639  :   DOI  :   10.1016/j.syapm.2015.02.007    
Abstract >>
Although the biogeography of rhizobia has been investigated extensively, little is known about the adaptive molecular evolution of rhizobia influenced by soil environments and selected by legumes. In this study, microevolution of Mesorhizobium strains nodulating Caragana in a semi-fixing desert belt in northern China was investigated. Five core genes-atpD, glnII, gyrB, recA, and rpoB, six heat-shock factor genes-clpA, clpB, dnaK, dnaJ, grpE, and hlsU, and five nodulation genes-nodA, nodC, nodD, nodG, and nodP, of 72 representative mesorhizobia were studied in order to determine their genetic variations. A total of 21 genospecies were defined based on the average nucleotide identity (ANI) of concatenated core genes using a threshold of 96% similarity, and by the phylogenetic analyses of the core/heat-shock factor genes. Significant genetic divergence was observed among the genospecies in the semi-fixing desert belt (areas A-E) and Yunnan province (area F), which was closely related to the environmental conditions and geographic distance. Gene flow occurred more frequently among the genospecies in areas A-E, and three sites in area B, than between area F and the other five areas. Recombination occurred among strains more frequently for heat-shock factor genes than the other genes. The results conclusively showed that the Caragana-associated mesorhizobia had divergently evolved according to their geographic distribution, and have been selected not only by the environmental conditions but also by the host plants.
KeywordMeSH Terms
Caragana
Mesorhizobium
Microevolution
Mutation
Phylogeny
Recombination
Caragana
Mesorhizobium
Microevolution
Mutation
Phylogeny
Recombination
Caragana
Mesorhizobium
Microevolution
Mutation
Phylogeny
Recombination
Gene Flow
Genetic Variation
Genotype
22. Uechi  K, Sakuraba  H, Yoshihara  A, Morimoto  K, Takata  G,     ( 2013 )

Structural insight into L-ribulose 3-epimerase from Mesorhizobium loti.

Acta crystallographica. Section D, Biological crystallography 69 (Pt 12)
PMID : 24311575  :   DOI  :   10.1107/S0907444913021665    
Abstract >>
L-Ribulose 3-epimerase (L-RE) from Mesorhizobium loti has been identified as the first ketose 3-epimerase that shows the highest observed activity towards ketopentoses. In the present study, the crystal structure of the enzyme was determined to 2.7 ? resolution. The asymmetric unit contained two homotetramers with the monomer folded into an (�\/�])8-barrel carrying four additional short �\-helices. The overall structure of M. loti L-RE showed significant similarity to the structures of ketose 3-epimerases from Pseudomonas cichorii, Agrobacterium tumefaciens and Clostridium cellulolyticum, which use ketohexoses as preferred substrates. However, the size of the C-terminal helix (�\8) was much larger in M. loti L-RE than the corresponding helices in the other enzymes. In M. loti L-RE the �\8 helix and the following C-terminal tail possessed a unique subunit-subunit interface which promoted the formation of additional intermolecular interactions and strengthened the enzyme stability. Structural comparisons revealed that the relatively small hydrophobic pocket of the enzyme around the substrate was likely to be the main factor responsible for the marked specificity for ketopentoses shown by M. loti L-RE.
KeywordMeSH Terms
d-psicose 3-epimerase
d-tagatose 3-epimerase
l-ribulose 3-epimerase
thermostability
23. Moukoumi  J, Hynes  RK, Dumonceaux  TJ, Town  J, Bélanger  N,     ( 2013 )

Characterization and genus identification of rhizobial symbionts from Caragana arborescens in western Canada.

Canadian journal of microbiology 59 (6)
PMID : 23750954  :   DOI  :   10.1139/cjm-2013-0158    
Abstract >>
Naturally occurring nitrogen-fixing symbionts from root nodules of caragana (Caragana arborescens) growing in central Saskatchewan were isolated following surface sterilization of caragana root nodules and squashing and spreading of the contents on yeast extract - mannitol medium. The symbiotic nature of the strains was confirmed following inoculation onto surface-sterilized C. arborescens seed in a gnotobiotic Leonard jar system. The Rhizobium isolates from C. arborescens root nodules were intermediate in generation time (g) (mean g of 5 isolates was 6.41 h) compared with the fast growers, Rhizobium leguminosarum NRG457 (g: 4.44 h), Rhizobium tropici 899 (g: 3.19 h), and Sinorhizobium meliloti BALSAC (g: 3.45 h), but they were faster than the slow-growing Bradyrhizobium japonicum USDA 110 (g: 13.86 h) and similar to Mesorhizobium amorphae (g: 7.76 h). Nitrogen derived from fixation by measuring changes in �_(15)N natural abundance in plant tissue confirmed the effectiveness of the strains; approximately 80% N2 from fixation. Strain identification was carried out by determining the sequences of 3 genes: 16S rRNA-encoding genes, cpn60, and recA. This analysis determined that the symbiotic partner of Canadian C. arborescens belongs to the genus Mesorhizobium and seems more related to M. loti than to previously described caragana symbionts like M. caraganae. This is the first report of Mesorhizobium sp. nodulating C. arborescens in western Canada.
KeywordMeSH Terms
Symbiosis
24. Nascimento  FX, Brígido  C, Glick  BR,     ( 2012 )

ACC deaminase genes are conserved among Mesorhizobium species able to nodulate the same host plant.

FEMS microbiology letters 336 (1)
PMID : 22846039  :   DOI  :   10.1111/j.1574-6968.2012.02648.x    
Abstract >>
Rhizobia strains expressing the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase have been reported to display an augmented symbiotic performance as a consequence of lowering the plant ethylene levels that inhibit the nodulation process. Genes encoding ACC deaminase (acdS) have been studied in Rhizobium spp.; however, not much is known about the presence of acdS genes in Mesorhizobium spp. The aim of this study was to assess the prevalence and phylogeny of acdS genes in Mesorhizobium strains including a collection of chickpea-nodulating mesorhizobia from Portugal. ACC deaminase genes were detected in 10 of 12 mesorhizobia type strains as well as in 18 of 18 chickpea Mesorhizobium isolates studied in this work. No ACC deaminase activity was detected in any Mesorhizobium strain tested under free-living conditions. Despite the lack of ACC deaminase activity, it was possible to demonstrate that in Mesorhizobium ciceri UPM-Ca7(T) , the acdS gene is transcribed under symbiotic conditions. Phylogenetic analysis indicates that strains belonging to different species of Mesorhizobium, but nodulating the same host plant, have similar acdS genes, suggesting that acdS genes are horizontally acquired by transfer of the symbiosis island. This data, together with analysis of the symbiosis islands from completely sequenced Mesorhizobium genomes, suggest the presence of the acdS gene in a Mesorhizobium common ancestor that possessed this gene in a unique symbiosis island.
KeywordMeSH Terms
Evolution, Molecular
Plant Root Nodulation
25. Kasai-Maita  H, Hirakawa  H, Nakamura  Y, Kaneko  T, Miki  K, Maruya  J, Okazaki  S, Tabata  S, Saeki  K, Sato  S,     ( 2013 )

Commonalities and differences among symbiosis islands of three Mesorhizobium loti strains.

Microbes and environments 28 (2)
PMID : 23666538  :   DOI  :   10.1264/jsme2.me12201     PMC  :   PMC4070662    
Abstract >>
To shed light on the breadth of the host range of Mesorhizobium loti strain NZP2037, we determined the sequence of the NZP2037 symbiosis island and compared it with those of strain MAFF303099 and R7A islands. The determined 533 kb sequence of NZP2037 symbiosis island, on which 504 genes were predicted, implied its integration into a phenylalanine-tRNA gene and subsequent genome rearrangement. Comparative analysis revealed that the core regions of the three symbiosis islands consisted of 165 genes. We also identified several NZP2037-specific genes with putative functions in nodulation-related events, suggesting that these genes contribute to broaden the host range of NZP2037.
KeywordMeSH Terms
Genetic Variation
Genomic Islands
26. Laranjo  M, Young  JP,     ( 2012 )

Multilocus sequence analysis reveals multiple symbiovars within Mesorhizobium species.

Systematic and applied microbiology 35 (6)
PMID : 22817876  :   DOI  :   10.1016/j.syapm.2012.06.002    
Abstract >>
The genus Mesorhizobium includes species nodulating several legumes, such as chickpea, which has a high agronomic importance. Chickpea rhizobia were originally described as either Mesorhizobium ciceri or M. mediterraneum. However, rhizobia able to nodulate chickpea have been shown to belong to several different species within the genus Mesorhizobium. The present study used a multilocus sequence analysis approach to infer a high resolution phylogeny of the genus Mesorhizobium and to confirm the existence of a new chickpea nodulating genospecies. The phylogenetic structure of the Mesorhizobium clade was evaluated by sequence analysis of the 16S rRNA gene, ITS region and the five core genes atpD, dnaJ, glnA, gyrB, and recA. Phylogenies obtained with the different genes are in overall good agreement and a well-supported, almost fully resolved, phylogenetic tree was obtained using the combined data. Our phylogenetic analyses of core genes sequences and their comparison with the symbiosis gene nodC, corroborate the existence of one new chickpea Mesorhizobium genospecies and one new symbiovar, M. opportunistum sv. ciceri. Furthermore, our results show that symbiovar ciceri spreads over six species of mesorhizobia. To our knowledge this study shows the most complete Mesorhizobium multilocus phylogeny to date and contributes to the understanding of how a symbiovar may be present in different species.
KeywordMeSH Terms
27.     ( 1996 )

Novel and complex chromosomal arrangement of Rhizobium loti nodulation genes.

Molecular plant-microbe interactions : MPMI 9 (3)
PMID : 8850088  :  
Abstract >>
A mutational and structural analysis of Rhizobium loti nodulation genes in strains NZP2037 and NZP2213 was carried out. Unlike the case with other Rhizobium strains examined to date, nodB was found on an operon separate from nodACIJ. Sequence analysis of the nodACIJ and nodB operon regions confirm that R. loti common nod genes have a gene organization different from that of other Rhizobium spp. At least 4 copies of nodD-like sequences were identified in R. loti. The complete nucleotide sequence of one of these, nodD3, was determined. A new host-specific nod gene, nolL, was identified adjacent to nodD3. NolL shares homology with NodX and other O-acetyl transferases. Mutational analysis of the nod regions of strains NZP2037 and NZP2213 showed that nodD3, nodI, nodJ, and nolL were all essential for R. loti strains to effectively nodulate the extended host Lotus pedunculatus, but were not necessary for effective nodulation of the less restrictive host, Lotus corniculatus. Both nodD3 and nolL were essential for R. loti strains to nodulate Leucaena leucocephala.
KeywordMeSH Terms
Genes, Bacterial
28. López-Lara  IM, van den Berg  JD, Thomas-Oates  JE, Glushka  J, Lugtenberg  BJ, Spaink  HP,     ( 1995 )

Structural identification of the lipo-chitin oligosaccharide nodulation signals of Rhizobium loti.

Molecular microbiology 15 (4)
PMID : 7783635  :   DOI  :   10.1111/j.1365-2958.1995.tb02372.x    
Abstract >>
Rhizobium loti is a fast-growing Rhizobium species that has been described as a microsymbiont of plants of the genus Lotus. Nodulation studies show that Lotus plants are nodulated by R. loti, but not by most other Rhizobium strains, indicating that R. loti produces specific lipo-chitin oligosaccharides (LCOs) which are necessary for the nodulation of Lotus plants. The LCOs produced by five different Rhizobium loti strains have been purified and were shown to be N-acetylglucosamine pentasaccharides of which the non-reducing residue is N-methylated and N-acylated with cis-vaccenic acid (C18:1) or stearic acid (C18:O) and carries a carbamoyl group. In one R. loti strain, NZP2037, an additional carbamoyl group is present on the non-reducing terminal residue. The major class of LCO molecules is substituted on the reducing terminal residue with 4-O-acetylfucose. Addition of LCOs to the roots of Lotus plants results in abundant distortion, swelling and branching of the root hairs, whereas spot inoculation leads to the formation of nodule primordia.
KeywordMeSH Terms
Plants, Medicinal
29.     ( 2013 )

Crystal structure of pyridoxine 4-oxidase from Mesorhizobium loti.

Biochimica et biophysica acta 1834 (6)
PMID : 23501672  :   DOI  :   10.1016/j.bbapap.2013.03.004    
Abstract >>
Pyridoxine 4-oxidase (PNOX) from Mesorhizobium loti is a monomeric glucose-methanol-choline (GMC) oxidoreductase family enzyme, catalyzes FAD-dependent oxidation of pyridoxine (PN) into pyridoxal, and is the first enzyme in pathway I for the degradation of PN. The tertiary structures of PNOX with a C-terminal His6-tag and PNOX-pyridoxamine (PM) complex were determined at 2.2? and at 2.1? resolutions, respectively. The overall structure consisted of FAD-binding and substrate-binding domains. In the active site, His460, His462, and Pro504 were located on the re-face of the isoalloxazine ring of FAD. PM binds to the active site through several hydrogen bonds. The side chains of His462 and His460 are located at 2.7 and 3.1? from the N4' atom of PM. The activities of His460Ala and His462Ala mutant PNOXs were very low, and 460Ala/His462Ala double mutant PNOX exhibited no activity. His462 may act as a general base for the abstraction of a proton from the 4'-hydroxyl of PN. His460 may play a role in the binding and positioning of PN. The C4' atom in PM is located at 3.2?, and the hydride ion from the C4' atom may be transferred to the N5 atom of the isoalloxazine ring. The comparison of active site residues in GMC oxidoreductase shows that Pro504 in PNOX corresponds to Asn or His of the conserved His-Asn or His-His pair in other GMC oxidoreductases. The function of the novel proline residue was discussed.
KeywordMeSH Terms
30. Marcos-García  M, Menéndez  E, Ramírez-Bahena  MH, Mateos  PF, Peix  ?, Velazquez  E, Rivas  R,     ( 2017 )

Mesorhizobium helmanticense sp. nov., isolated from Lotus corniculatus nodules.

International journal of systematic and evolutionary microbiology 67 (7)
PMID : 28693668  :   DOI  :   10.1099/ijsem.0.001942    
Abstract >>
In this study, three strains belonging to the genus Mesorhizobium, CSLC115NT, CSLC19N and CSLC37N, isolated from Lotus corniculatus nodules in Spain, were characterized. Their 16S rRNA gene sequences were closely related to those of Mesorhizobium metallidurans STM 2683T, Mesorhizobium tianshanense A-1BST, Mesorhizobium tarimense CCBAU 83306T, Mesorhizobium gobiense CCBAU 83330T and Mesorhizobium caraganae CCBAU 11299T with similarity values higher than 99.7 %. The analysis of concatenated recA and glnII genes showed that the most closely related type strains were M. metallidurans STM 2683T, M. tianshanense A-1BST and M. tarimense CCBAU 83306T with 96, 95 and 94 % similarity values in the recA gene and 95, 94 and 94 % in the glnII gene, respectively. M. metallidurans LMG 24485T, M. tianshanense USDA 3592T and M. tarimense LMG 24338T showed means of 44, 41 and 42 % DNA-DNA relatedness, respectively, with respect to strain CSLC115NT. The major fatty acids were those from summed feature 8 (C18 : 1�s7c/C18 : 1�s6c), C16 : 0 and C18 : 1�s7c 11-methyl. The results of phenotypic characterization support that the L. corniculatus nodulating strains analysed in this work belong to a novel species of the genus Mesorhizobium for which the name Mesorhizobium helmanticense sp. nov. is proposed, and the type strain is CSLC115NT (= LMG 29734T=CECT 9168T).
KeywordMeSH Terms
Phylogeny
31.     ( 2013 )

Mesorhizobium qingshengii sp. nov., isolated from effective nodules of Astragalus sinicus.

International journal of systematic and evolutionary microbiology 63 (Pt 6)
PMID : 23041644  :   DOI  :   10.1099/ijs.0.044362-0    
Abstract >>
In a study on the diversity of rhizobia isolated from root nodules of Astragalus sinicus, five strains showed identical 16S rRNA gene sequences. They were related most closely to the type strains of Mesorhizobium loti, Mesorhizobium shangrilense, Mesorhizobium ciceri and Mesorhizobium australicum, with sequence similarities of 99.6-99.8%. A polyphasic approach, including 16S-23S intergenic spacer (IGS) RFLP, comparative sequence analysis of 16S rRNA, atpD, glnII and recA genes, DNA-DNA hybridization and phenotypic tests, clustered the five isolates into a coherent group distinct from all recognized Mesorhizobium species. Except for strain CCBAU 33446, from which no symbiotic gene was detected, the four remaining strains shared identical nifH and nodC gene sequences and nodulated with Astragalus sinicus. In addition, these five strains showed similar but different fingerprints in IGS-RFLP and BOX-repeat-based PCR, indicating that they were not clones of the same strain. They were also distinguished from recognized Mesorhizobium species by several phenotypic features and fatty acid profiles. Based upon all the results, we suggest that the five strains represent a novel species for which the name Mesorhizobium qingshengii sp. nov. is proposed. The type strain is CCBAU 33460(T) (=CGMCC 1.12097(T)=LMG 26793(T)=HAMBI 3277(T)). The DNA G+C content of the type strain is 59.52 mol% (Tm).
KeywordMeSH Terms
Phylogeny
32.     ( 2012 )

Robust markers reflecting phylogeny and taxonomy of rhizobia.

PloS one 7 (9)
PMID : 23028691  :   DOI  :   10.1371/journal.pone.0044936     PMC  :   PMC3444505    
Abstract >>
Genomic ANI (Average Nucleotide Identity) has been found to be able to replace DNA-DNA hybridization in prokaryote taxonomy. The ANI of each of the core genes that has a phylogeny congruent with the reference species tree of rhizobia was compared to the genomic ANI. This allowed us to identify three housekeeping genes (SMc00019-truA-thrA) whose ANI reflected the intraspecies and interspecies genomic ANI among rhizobial strains, revealing an ANI gap (?2%) between the inter- and intra-species comparisons. The intraspecies (96%) and interspecies (94%) ANI boundaries calculated from three genes (SMc00019-truA-thrA) provided a criterion for bacterial species definition and confirmed 621/629 of known interspecies relationships within Bradyrhizobium, Mesorhizobium, Sinorhizobium and Rhizobium. Some widely studied strains should be renamed. The SMc00019-truA-thrA ANI also correlates well with the genomic ANI of strains in Agrobacterium, Methylobacterium, Ralstonia, Rhodopseudomonas, Cupriavidus and Burkholderia, suggesting their wide applicability in other bacteria.
KeywordMeSH Terms
Phylogeny
33.     ( 1998 )

Evolution of rhizobia by acquisition of a 500-kb symbiosis island that integrates into a phe-tRNA gene.

Proceedings of the National Academy of Sciences of the United States of America 95 (9)
PMID : 9560243  :   DOI  :   10.1073/pnas.95.9.5145     PMC  :   PMC20228    
Abstract >>
Nodulation and nitrogen fixation genes of Mesorhizobium loti are encoded on the chromosome of the bacterium. Nevertheless, there is strong evidence that these genes can be transferred from an inoculant strain to nonsymbiotic mesorhizobia in the field environment. Here we report that the chromosomal symbiotic element of M. loti strain ICMP3153 is transmissible in laboratory matings to at least three genomic species of nonsymbiotic mesorhizobia. The element is 500 kb in size, integrates into a phe-tRNA gene, and encodes an integrase of the phage P4 family just within its left end. The entire phe-tRNA gene is reconstructed at the left end of the element upon integration, whereas the 3' 17 nucleotides of the tRNA gene are present as a direct repeat at the right end. We termed the element a symbiosis island on the basis of its many similarities to pathogenicity islands. It may represent a class of genetic element that contributes to microbial evolution by acquisition.
KeywordMeSH Terms
Biological Evolution
Symbiosis

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