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      rpsU-based discrimination within the genus Burkholderia

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          Abstract

          Sequencing of the gene rpsU reliably delineates saprophytic Burkholderia ( B.) thailandensis from highly pathogenic B. mallei and B. pseudomallei. We analyzed the suitability of this technique for the delineation of the B. pseudomallei complex from other Burk-holderia species.

          Both newly recorded and previously deposited sequences of well-characterized or reference strains ( n = 84) of Azoarcus spp., B. ambifaria, B. anthina, B. caledonica, B. caribensis, B. caryophylli, B. cenocepacia, B. cepacia, B. cocovenenans, B. dolosa, B. fungorum, B. gladioli, B. glathei, B. glumae, B. graminis, B. hospita, B. kururensis, B. mallei, B. multivorans, B. phenazinium, B. phenoliruptrix, B. phymatum, B. phytofirmans, B. plantarii, B. pseudomallei, B. pyrrocinia, B. stabilis, B. thailandensis, B. ubonensis, B. vietnamiensis, B. xenovorans, not further defined Burkholderia spp., and the outliers Cupriavidus metallidurans, Laribacter hongkongensis, Pandorea norimbergensis, and Ralstonia pickettii were included in a multiple sequence analysis.

          Multiple sequence alignments led to the delineation of four major clusters, rpsU-I to rpsU-IV, with a sequence homology >92%. The B. pseudomallei complex formed the complex rpsU-II. Several Burkholderia species showed 100% sequence homology.

          This procedure is useful for the molecular confirmation or exclusion of glanders or melioidosis from primary patient material. Further discrimination within the Burkholderia genus requires other molecular approaches.

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          Most cited references52

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          Taxonomy of the genus Cupriavidus: a tale of lost and found.

          DNA-DNA hybridization experiments and an evaluation of phenotypic characteristics, DNA base ratios and 16S rRNA gene sequences demonstrated that Wautersia eutropha (Davies 1969) Vaneechoutte et al. 2004, the type species of the genus Wautersia, is a later synonym of Cupriavidus necator Makkar and Casida 1987, the type species of the genus Cupriavidus. In conformity with Rules 15, 17, 23a and 37a(1) of the International Code of Nomenclature of Bacteria, the genus name Cupriavidus has priority over the genus name Wautersia, and all other members of the genus Wautersia are reclassified into Cupriavidus as Cupriavidus basilensis comb. nov. (type strain LMG 18990(T)=DSM 11853(T)), Cupriavidus campinensis comb. nov. (type strain LMG 19282(T)=CCUG 44526(T)), Cupriavidus gilardii comb. nov. (type strain LMG 5886(T)=CCUG 38401(T)), Cupriavidus metallidurans comb. nov. (type strain LMG 1195(T)=DSM 2839(T)), Cupriavidus oxalaticus comb. nov. (type strain LMG 2235(T)=CCUG 2086(T)=DSM 1105(T)), Cupriavidus pauculus comb. nov. (type strain LMG 3244(T)=CCUG 12507(T)), Cupriavidus respiraculi comb. nov. (type strain LMG 21510(T)=CCUG 46809(T)) and Cupriavidus taiwanensis comb. nov. (type strain LMG 19424(T)=CCUG 44338(T)).
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            Burkholderia phytofirmans sp. nov., a novel plant-associated bacterium with plant-beneficial properties.

            A Gram-negative, non-sporulating, rod-shaped, motile bacterium, with a single polar flagellum, designated strain PsJN(T), was isolated from surface-sterilized onion roots. This isolate proved to be a highly effective plant-beneficial bacterium, and was able to establish rhizosphere and endophytic populations associated with various plants. Seven related strains were recovered from Dutch soils. Based on 16S rRNA gene sequence data, strain PsJN(T) and the Dutch strains were identified as representing a member of the genus Burkholderia, as they were closely related to Burkholderia fungorum (98.7 %) and Burkholderia phenazinium (98.5 %). Analysis of whole-cell protein profiles and DNA-DNA hybridization experiments confirmed that all eight strains belonged to a single species. Strain PsJN(T) had a DNA G+C content of 61.0 mol%. Only low levels of DNA-DNA hybridization to closely related species were found. Qualitative and quantitative differences in fatty acid composition between strain PsJN(T) and closely related species were identified. The predominant fatty acids in strain PsJN(T) were 16 : 0, 18 : 1omega7c and summed feature 3 (comprising 16 : 1omega7c and/or iso-15 : 0 2-OH). Isolate PsJN(T) showed high 1-aminocyclopropane-1-carboxylate deaminase activity and is therefore able to lower the ethylene level in a developing or stressed plant. Production of the quorum-sensing signal compound 3-hydroxy-C8-homoserine lactone was detected. Based on the results of this polyphasic taxonomic study, strain PsJN(T) and the seven Dutch isolates are considered to represent a single, novel species, for which the name Burkholderia phytofirmans sp. nov. is proposed. The type strain is strain PsJN(T) (=LMG 22146(T) = CCUG 49060(T)).
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              Melioidosis.

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                Author and article information

                Journal
                1886
                122234
                European Journal of Microbiology and Immunology
                EuJMI
                Akadémiai Kiadó, co-published with Springer Science+Business Media B.V., Formerly Kluwer Academic Publishers B.V.
                2062-509X
                2062-8633
                1 June 2014
                : 4
                : 2
                : 106-116
                Affiliations
                [ 1 ] Department of Tropical Medicine at the Bernhard Nocht Institute, German Armed Forces Hospital of Hamburg, Bernhard Nocht street 74, D-20359, Hamburg, Germany
                [ 2 ] Institute for Microbiology, Virology and Hygiene, University Hospital of Rostock, Rostock, Germany
                [ 3 ] Friedrich Loeffler Institute, Federal Research Institute for Animal Health, Jena, Germany
                [ 4 ] University Medicine Goettingen, Goettingen, Germany
                Author notes

                Supplementary material is available for this article at 10.1556/EuJMI.4.2014.2.3 and is accessible for authorized users.

                [* ] 0049-40-6947-28743, 0049-40-6947-28709, Frickmann@ 123456bni-hamburg.de
                Article
                3
                10.1556/eujmi.4.2014.2.3
                4029289
                24883196
                77a1dc34-2b7d-45b7-b8d4-a36befe35e6f
                History
                : 27 March 2014
                : 8 April 2014
                Categories
                Original Article

                Medicine,Immunology,Health & Social care,Microbiology & Virology,Infectious disease & Microbiology
                discrimination, Burkholderia , rpsU ,sequencing,ribosomal subunit protein S21

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