RFLP of rRNA genes and sequencing of the 16S-23S rDNA intergenic spacer region of ammonia-oxidizing bacteria: a phylogenetic approach Free

Abstract

It has been established that 16S rRNA gene-based phytogeny gives a low resolution between members of the chemoautotrophic ammonia-oxidizing bacteria (AOB) belonging to the -subclass of the . In this study, 12 isolates of AOB were ribotyped, and the sequences of the 16S–23S rDNA intergenic spacer region (ISR) were determined and used in a phylogenetic study. 16S and 23S rDNA ribotyping revealed that the AOB studied contain only one operon per genome, in contrast to most bacteria, which have 5–10 copies of the rRNA genes per genome. It is likely that the presence of only one set of rRNA genes is related to the slow growth of the AOB. The 16S and 23S rRNA genes of the AOB were shown to be arranged in the classical way: a 16S rRNA gene, an ISR and a 23S rRNA gene. Despite the close phylogenetic relationship among the AOB, the relative location of the rRNA genes in the genome appears to vary considerably. The size of the ISR was approximately 400 bp in the isolates and 645–694 bp in the isolates, suggesting a species-specific size difference in the ISR. The ISR contained two potential tRNA genes in the 5′ end in all isolates studied. The similarity values between the ISR sequences of the AOB are low (42·9–96·2%) compared with the 16S rDNA sequence similarity values, and therefore the ISR sequences are valuable as a complementary phylogenetic tool in combination with 16S rRNA gene sequences. The phylogenetic analysis of the AOB based on ISR sequences confirms the 16S rRNA gene-based phytogeny but has the benefit of giving a higher resolution.

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1999-01-01
2024-03-28
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