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PubMed2004

Rice cDNA microarray-based gene expression profiling of the response to flagellin perception in cultured rice cells.

Incompatible strains of Acidovorax avenae elicit an immune response in cultured rice cells, with immunity specifically induced by the flagellin of the incompatible strain. To identify genes regulated by flagellin perception signaling in cultured rice cells, gene expression patterns were analyzed with rice cDNA microarrays, including 3,353 independent rice cDNA clones. In all, 131 genes were differentially expressed between incompatible and compatible interactions. K-means clustering showed that 94 genes were upregulated and 32 genes were downregulated during incompatible interactions, whereas only 5 genes were upregulated during compatible interactions. Among the 126 genes that were up- or downregulated during incompatible interactions, expression of 46 genes was decreased when cultured rice cells were inoculated with a flagellin-deficient incompatible strain (delta fla1141-2), indicating that approximately 37% of the 126 genes were directly controlled by flagellin perception. Real-time reverse-transcription polymerase chain reaction analysis using flagellins purified from incompatible or compatible strains was performed to confirm flagellin-regulated expression of candidate genes selected by microarray analysis. Results showed that induction of some genes involved in the immune response is regulated not only by the flagellin perception pathway, but also by another recognition molecule-perception pathway.

Bacterial Proteins↗
PubMed2003

Caenibacterium thermophilum gen. nov., sp. nov., isolated from a thermophilic aerobic digester of municipal sludge.

A bacterial strain, N2-680(T) (=DSM 15264(T)=LMG 21760(T)), isolated from a thermophilic aerobic digester of municipal sludge, was characterized with respect to its morphology, physiology and taxonomy. Phenotypically, the isolate was a Gram-negative rod with a polar flagellum, catalase- and oxidase-positive, containing cytoplasmic inclusions of poly-beta-hydroxybutyrate and had an optimal growth temperature of about 47 degrees C. Strain N2-680(T) was unable to reduce nitrate and could use organic acids, amino acids and carbohydrates as single carbon sources. Chemotaxonomic analysis revealed that ubiquinone 8 was the major respiratory quinone of this organism and that phosphatidylethanolamine and phosphatidylglycerol were the major polar lipids. At 50 degrees C, the major components in fatty acid methyl ester analysis were C(16 : 0) and cyclo-C(17 : 0). The highest 16S rDNA sequence identity of isolate N2-680(T) was to Leptothrix mobilis and Ideonella dechloratans (95.7 %) and to Rubrivivax gelatinosus and Aquabacterium commune (95.6 %). 16S rDNA sequence similarities to species of two related thermophilic genera, Caldimonas manganoxidans and Tepidimonas ignava, were lower (93.6 and 94.7 %). On the basis of phylogenetic analyses and physiological and chemotaxonomic characteristics, it is proposed that isolate N2-680(T) represents a new genus and species, for which the name Caenibacterium thermophilum gen. nov., sp. nov. is proposed.

Comamonadaceae↗
PubMed2004

Emended description of the species Lampropedia hyalina.

Three Lampropedia hyalina strains from different habitats were compared by phenotypic, chemotaxonomic and molecular characteristics. All strains form coccoid cells and have been reported to grow as square tablets of eight to 64 cells. However, two of these strains (ATCC 11041T and ATCC 43383) have apparently lost this ability, and the third strain may temporarily lose this capacity under certain cultivation conditions. The three strains showed only minor differences in metabolic characteristics: the main significant physiological difference was the ability to accumulate polyphosphate under alternating anaerobic-aerobic conditions found for DSM 15336. The three strains showed high similarity in fatty acid composition and only slight differences in the G + C content (63-67 mol%) and DNA-DNA reassociation (90-95 % relatedness). Comparative 16S rRNA gene sequence analyses on these three strains and three Lampropedia hyalina 16S rRNA gene sequences deposited at NCBI showed that they are all very similar (> 98.8 %) and that they form a distinct group among the 'Betaproteobacteria', showing between 94.6 and 93 % 16S rRNA gene similarity to members of various genera such as Acidovorax, Aquaspirillum, Brachymonas, Comamonas, Delftia and Xenophilus. Fluorescent in situ hybridization with oligonucleotide probes targeting betaproteobacteria on the 16S rRNA and 23S rRNA gene level further supported the conclusion that all investigated strains are members of the 'Betaproteobacteria'. Two oligonucleotide probes were designed and successfully applied for culture-independent identification of Lampropedia hyalina by means of fluorescent in situ hybridization.

Aerobiosis↗
PubMed2004

Proposals of Curvibacter gracilis gen. nov., sp. nov. and Herbaspirillum putei sp. nov. for bacterial strains isolated from well water and reclassification of [Pseudomonas] huttiensis, [Pseudomonas] lanceolata, [Aquaspirillum] delicatum and [Aquaspirillum] autotrophicum as Herbaspirillum huttiense comb. nov., Curvibacter lanceolatus comb. nov., Curvibacter delicatus comb. nov. and Herbaspirillum autotrophicum comb. nov.

Two strains of curved bacteria, 7-1(T) and 7-2(T), isolated from well water, were phylogenetically examined to determine their taxonomic position. Strain 7-1(T) is a Gram-negative, slightly curved rod. Analysis of the 16S rRNA gene sequence showed that strain 7-1(T) formed a cluster with [Aquaspirillum] delicatum and [Pseudomonas] lanceolata. It has some similar characteristics to [A.] delicatum and [P.] lanceolata, but has sufficient distance to separate it from other genera. DNA-DNA hybridization analysis, as well as chemotaxonomic and morphological studies, demonstrated that strain 7-1(T), [A.] delicatum and [P.] lanceolata belong to a new genus, Curvibacter gen. nov. Strain 7-1(T) (=IAM 15033(T)=ATCC BAA-807(T)) is classified as the type strain of Curvibacter gracilis gen. nov., sp. nov., and [A.] delicatum and [P.] lanceolata are classified as Curvibacter delicatus comb. nov. and Curvibacter lanceolatus comb. nov., respectively. Strain 7-2(T) is a Gram-negative spirillum. Phylogenetic study based on the 16S rRNA gene sequences showed that it formed a cluster with the members of the genus Herbaspirillum, [Pseudomonas] huttiensis and [Aquaspirillum] autotrophicum. The classification is therefore proposed of strain 7-2(T) (=IAM 15032(T)=ATCC BAA-806(T)) as the type strain of Herbaspirillum putei sp. nov., and [P.] huttiensis and [A.] autotrophicum are transferred to the genus Herbaspirillum as Herbaspirillum huttiense comb. nov. and Herbaspirillum autotrophicum comb. nov., respectively.

Bacterial Typing Techniques↗
PubMed2004

Caenibacterium thermophilum is a later synonym of Schlegelella thermodepolymerans.

Recently, two strains of Schlegelella thermodepolymerans Elbanna et al. 2003 and an independently isolated bacterium, Caenibacterium thermophilum Manaia et al. 2003, were described in parallel as gen. nov., sp. nov. Analysis of the 16S rRNA genes revealed similarity between C. thermophilum and the two strains of S. thermodepolymerans of 99.8 and 99.6 %, respectively. DNA-DNA hybridization experiments revealed mean DNA reassociation levels of 97-98 % among C. thermophilum and the two strains of S. thermodepolymerans, thereby confirming the close relationship and indicating that C. thermophilum is a later synonym of S. thermodepolymerans.

Comamonadaceae↗
PubMed2005

Reclassification of Alcaligenes latus strains IAM 12599T and IAM 12664 and Pseudomonas saccharophila as Azohydromonas lata gen. nov., comb. nov., Azohydromonas australica sp. nov. and Pelomonas saccharophila gen. nov., comb. nov., respectively.

The aim of this study was to clarify the taxonomic position of the nitrogen-fixing and hydrogen-oxidizing bacteria Alcaligenes latus strains IAM 12599T, IAM 12664 and IAM 12665 and Pseudomonas saccharophila IAM 14368T. It was found that the type strain of Alcaligenes latus, IAM 12599T, showed 99 x 9 and 96 x 1 % 16S rRNA gene sequence similarity to strains IAM 12665 and IAM 12664, respectively. A comparison using DNA-DNA hybridization suggested that strains IAM 12599T and IAM 12665 belong to a single species (89 x 7 %) and that strain IAM 12664 (35 x 1 %) forms a separate species. The phenotypic characteristics also support the conclusion that these bacteria should be identified as two species of a new genus: Azohydromonas lata gen. nov., comb. nov. (type strain IAM 12599T=DSM 1122T=LMG 3321T=ATCC 29712T; reference strain IAM 12665=DSM 1123=LMG 3325=ATCC 29714) and Azohydromonas australica sp. nov. (type strain IAM 12664T=DSM 1124T=LMG 3324T=ATCC 29713T). Pseudomonas saccharophila IAM 14368T was found to be closely related to the phototrophic bacterium Roseateles depolymerans, with 96 x 8 % 16S rRNA gene sequence similarity, but the two bacteria are quite different with respect to their metabolism and some significant phenotypic characteristics, suggesting that they cannot be included in a single genus. Further studies on their nifH gene sequences, G+C content of the DNA and cellular fatty acid composition confirm that Pseudomonas saccharophila should be reclassified: the name Pelomonas saccharophila gen. nov., comb. nov. is proposed, with the type strain IAM 14368T (=LMG 2256T=ATCC 15946T).

Alcaligenes↗
PubMed2006

Variovorax dokdonensis sp. nov., isolated from soil.

A Gram-negative, motile and oval- or rod-shaped bacterial strain, DS-43T, was isolated from soil from Dokdo, Korea, and its taxonomic position was investigated by using a polyphasic approach. The strain grew optimally at pH 7.0-8.0 and at 30 degrees C in the presence of 0.5 % (w/v) NaCl. Strain DS-43T had Q-8 as the predominant ubiquinone and C(16 : 0), C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH, C(18 : 1)omega7c and C(17 : 0) cyclo as the major fatty acids, which are consistent with the corresponding data for the type strain (KCTC 12459T) of Variovorax paradoxus. The DNA G+C content was 66.0 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain DS-43T was most closely related to Variovorax paradoxus, the only recognized species of the genus. The level of 16S rRNA gene sequence similarity between strain DS-43T and V. paradoxus IAM 12373T was 97.3 %. DNA-DNA relatedness data and differential phenotypic properties demonstrated that strain DS-43T is distinguishable from V. paradoxus. On the basis of phenotypic, phylogenetic and genetic data, it is proposed that strain DS-43T (=KCTC 12544T = CIP 108838T) be classified in the genus Variovorax as a member of a novel species, Variovorax dokdonensis sp. nov.

Comamonadaceae↗
PubMed2006

Variovorax soli sp. nov., isolated from greenhouse soil.

A Gram-negative, rod-shaped, non-spore-forming bacterium, strain GH9-3(T), isolated from greenhouse soil, was investigated in a polyphasic study. The novel organism grew at 10-35 degrees C, 0-3 % NaCl and pH 5-9. It had ubiquinone 8 (Q-8) as the predominant isoprenoid quinone and possessed C(16 : 0), summed feature 3, C(17 : 0) cyclo and C(18 : 1) omega7c as the major fatty acids (together representing 87.4 % of the total). The DNA G+C content was 67.1 mol%. 16S rRNA gene sequence analysis of strain GH9-3(T) showed that it grouped within the Variovorax cluster, with highest sequence similarities to Variovorax paradoxus IAM 12373(T) (98.3 %) and Variovorax dokdonensis DS-43(T) (98.0 %). DNA-DNA hybridization values between strain GH9-3(T) and V. paradoxus DSM 30034(T) and V. dokdonensis DS-43(T) were 38 and 29 %, respectively. Based on phenotypic, chemotaxonomic and phylogenetic features, it is proposed that strain GH9-3(T) represents a novel species of the genus Variovorax with the name Variovorax soli sp. nov. The type strain is GH9-3(T) (=KACC 11579(T)=DSM 18216(T)).

Bacterial Typing Techniques↗
PubMed2007

Simplicispira limi sp. nov., isolated from activated sludge.

A Gram-negative, motile, rod-shaped bacterium, designated strain EMB325(T), was isolated from activated sludge that performed enhanced biological phosphorus removal in a sequencing batch reactor. The predominant fatty acids of strain EMB325(T) were summed feature 3 (C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH), C(16 : 0), C(18 : 1)omega7c and C(18 : 1)omega7c 11-methyl. The strain contained phosphatidylethanolamine and diphosphatidylglycerol as polar lipids. The G+C content of the genomic DNA was 63.3 mol%. The major quinone was Q-8. Phylogenetic analysis of 16S rRNA gene sequences showed that strain EMB325(T) formed a phyletic lineage with members of the genus Simplicispira and was most closely related to Simplicispira psychrophila LMG 5408(T) and Simplicispira metamorpha DSM 1837(T) with similarities of 98.1 and 97.9 %, respectively. Levels of DNA-DNA relatedness between strain EMB325(T) and S. psychrophila LMG 5408(T) and S. metamorpha DSM 1837(T) were 28 and 23 %, respectively. On the basis of chemotaxonomic data and molecular characteristics, strain EMB325(T) is considered to represent a novel species within the genus Simplicispira, for which the name Simplicispira limi sp. nov. is proposed. The type strain is EMB325(T) (=KCTC 12608(T)=DSM 17964(T)).

Bacterial Typing Techniques↗
PubMed2006

Characterization of the genes encoding the 3-carboxy-cis,cis-muconate-lactonizing enzymes from the 4-sulfocatechol degradative pathways of Hydrogenophaga intermedia S1 and Agrobacterium radiobacter S2.

Hydrogenophaga intermedia strain S1 and Agrobacterium radiobacter strain S2 form a mixed bacterial culture which degrades sulfanilate (4-aminobenzenesulfonate) by a novel variation of the beta-ketoadipate pathway via 4-sulfocatechol and 3-sulfomuconate. It was previously proposed that the further metabolism of 3-sulfomuconate is catalysed by modified 3-carboxy-cis,cis-muconate-lactonizing enzymes (CMLEs) and that these 'type 2' enzymes were different from the conventional CMLEs ('type 1') from the protocatechuate pathway in their ability to convert 3-sulfomuconate in addition to 3-carboxy-cis,cis-muconate. In the present study the genes for two CMLEs (pcaB2S1 and pcaB2S2) were cloned from H. intermedia S1 and A. radiobacter S2, respectively. In both strains, these genes were located close to the previously identified genes encoding the 4-sulfocatechol-converting enzymes. The gene products of pcaB2S1 and pcaB2S2 were therefore tentatively identified as type 2 enzymes involved in the metabolism of 3-sulfomuconate. The genes were functionally expressed and the gene products were shown to convert 3-carboxy-cis,cis-muconate and 3-sulfomuconate. 4-Carboxymethylene-4-sulfo-but-2-en-olide (4-sulfomuconolactone) was identified by HPLC-MS as the product, which was enzymically formed from 3-sulfomuconate. His-tagged variants of both CMLEs were purified and compared with the CMLE from the protocatechuate pathway of Pseudomonas putida PRS2000 for the conversion of 3-carboxy-cis,cis-muconate and 3-sulfomuconate. The CMLEs from the 4-sulfocatechol pathway converted 3-sulfomuconate with considerably higher activities than 3-carboxy-cis,cis-muconate. Also the CMLE from P. putida converted 3-sulfomuconate, but this enzyme demonstrated a clear preference for 3-carboxy-cis,cis-muconate as substrate. Thus it was demonstrated that in the 4-sulfocatechol pathway, distinct CMLEs are formed, which are specifically adapted for the preferred conversion of sulfonated substrates.

Amino Acid Sequence↗
PubMed2006

Cloning and characterization of the chromosomal arsenic resistance genes from Acidithiobacillus caldus and enhanced arsenic resistance on conjugal transfer of ars genes located on transposon TnAtcArs.

All strains of the moderately thermophilic, acidophilic, sulphur-oxidizing bacterium Acidithiobacillus caldus that have been tested contain a set of chromosomal arsenic resistance genes. Highly arsenic-resistant strains isolated from commercial arsenopyrite bio-oxidation tanks contain additional transposon-located (TnAtcArs) arsenic resistance genes. The chromosomal At. caldus ars genes were cloned and found to consist of arsR and arsC genes transcribed in one direction, and arsB in the opposite direction. The arsRC genes were co-transcribed with ORF1, and arsB with ORF5 in both At. caldus and Escherichia coli, although deletion of ORFs 1 and 5 did not appear to affect resistance to arsenate or arsenite in E. coli. ORFs 1 and 5 have not previously been reported as part of the ars operons, and had high amino acid identity to hypothetical proteins from Polaromonas naphthalenivorus (76%) and Legionella pneumophila (60%), respectively. Reporter-gene studies showed that the arsenic operon of transposon origin (TnAtcArs) was expressed at a higher level, and was less tightly regulated in E. coli than were the At. caldus ars genes of chromosomal origin. Plasmid pSa-mediated conjugal transfer of TnAtcArs from E. coli to At. caldus strains lacking the transposon was successful, and resulted in greatly increased levels of resistance to arsenite.

Acidithiobacillus↗
PubMed2006

Identification of a response regulator gene for catabolite control from a PCB-degrading beta-proteobacteria, Acidovorax sp. KKS102.

Acidovorax sp. (formally Pseudomonas sp.) strain KKS102 carries a bph operon for the degradation of PCB/biphenyl. Transcription from the pE promoter for the bph operon was found to be under catabolite control, i.e. the promoter activity was at a lower level when succinate, fumarate or acetate was added to the culture. Some mutations in the immediate upstream region of the pE promoter resulted in catabolite-insensitive and constitutively low promoter activity, suggesting that a transcriptional activator was involved in catabolite control. A genetic screen for a pE promoter activator identified two tandemly arranged genes, bphP and bphQ, that encoded proteins homologous to the sensor kinases and response regulators, respectively, of two-component regulatory system. In the bphPQ double mutant, pE promoter activity was weak and catabolite-insensitive, and a supply of the bphQ gene alone led to the restoration of the catabolite response. The mechanism of catabolite repression in KKS102 is explained in terms of inhibition of activation by BphQ. The genes highly similar to bphQ were found from several beta-proteobacteria, such as Burkholderia cenocepacia J2315, B. multivorans ATCC17616, B. xenovorans LB400 and Ralstonia solanacearum RS1085.

Bacterial Proteins↗
PubMed2005

Proposed involvement of a soluble methane monooxygenase homologue in the cyclohexane-dependent growth of a new Brachymonas species.

High-throughput mRNA differential display (DD) was used to identify genes induced by cyclohexane in Brachymonas petroleovorans CHX, a recently isolated beta-proteobacterium that grows on cyclohexane. Two metabolic gene clusters were identified multiple times in independent reverse transcription polymerase chain reactions (RT-PCR) in the course of this DD experiment. These clusters encode genes believed to be required for cyclohexane metabolism. One gene cluster (8 kb) encodes the subunits of a multicomponent hydroxylase related to the soluble butane of Pseudomonas butanovora and methane monooxygenases (sMMO) of methanotrophs. We propose that this butane monooxygenase homologue carries out the oxidation of cyclohexane into cyclohexanol during growth. A second gene cluster (11 kb) contains almost all the genes required for the oxidation of cyclohexanol to adipic acid. Real-time PCR experiments confirmed that genes from both clusters are induced by cyclohexane. The role of the Baeyer-Villiger cyclohexanone monooxygenase of the second cluster was confirmed by heterologous expression in Escherichia coli.

Adipates↗