Use of Polymerase Chain Reaction-amplified 16S rRNA Gene Sequences to Identify Pink-pigmented Bacteria Found in a Potable Water Treatment System.

Abstract

Two strains of pink-pigmented, chlorine-resistant bacteria, GK101 and GK118, previously isolated from pink-colored microbial biomass occurring in a potable water treatment system were studied phylogenetically by using 16S rRNA gene sequence information. 16S ribosomal DNA (rDNA) fragments of approximately 1.5 kbp were amplified by the polymerase chain reaction (PCR) from crude cell lysates of the isolates and sequenced directly by a combined method consisting of linear PCR sequencing and automated fluorescence detection. Sequence comparisons and a phylogenetic tree analysis showed that the pink-pigmented isolates were closely related to Methylobacterium extorquens. Smaller fragments of 16S rDNA were also amplified from the bulk DNA extracted from the pink biomass, with a pair of PCR primers specific to the methylotrophs with the serine pathway or to proteobacteria of the alpha subclass. The PCR product was cloned by using the T-A cloning technique and sequenced as noted above. The results of this molecular approach suggested that bacteria phylogenetically the same as strain GK 101 predominated in the pink-pigmented population in situ and that some other bacteria of the genus Methylobacterium might also be involved in formation of the pink-colored aggregates.

Detection of Intracellular Bacteria in the Buds of Scotch Pine ( Pinus sylvestris L.) by In Situ Hybridization

“…Figure 3 contains two neighborjoining trees showing the phylogenetic positions of bacterial isolates F and G. Isolate F formed a cluster with Methylobacterium sp. strain GK 101 (14), supported by a bootstrap value of 899. Isolate G formed a cluster with P. azotoformans, P. synxantha, and P. mucidolens, supported by a bootstrap value of 989.…”

Phenotypic and genetic diversity of chlorine-resistant Methylobacterium strains isolated from various environments

“…They are phenotypically and chemotaxonomically quite similar, and phenotypic differences among the species are found in only limited properties, such as carbon source utilization. In recent years, 16S rRNA sequence information has been used for phylogenetic placement and identification of different physiological groups of methylotrophic proteobacteria, including Methylobacterium species (4,6,27,51,52). The molecular data have shown that the genus Methylobacterium represents a line of descent in the alpha-2 subclass of the class Proteobacteria.…”

“…Our approaches to this investigation were twofold: one involved physiological and biochemical characterization of the isolates, followed by numerical analysis of phenotypic profiles, and the other involved genetic analysis by PCR sequencing of 16S ribosomal DNA (rDNA). A previous phylogenetic study has shown that some Methylobacterium species, including those closely related to M. extorquens, are responsible for the formation of pink biofilms in a potable water treatment system (27).…”

Isolation and 16S DNA Characterization of Soil Microorganisms from Tropical Soils Capable of Utilizing the Herbicides Hexazinone and Tebuthiuron

“…[32] Of principal relevance to the present research, however, is the fact that Methylobacterium spp. have been isolated from soils.…”