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Abstract:
In order to investigate the genetic diversity of streptomycetes in an
acid forested soil sample from Mt. Coot-tha, Brisbane, Australia, cells
were mechanically lysed within the soil matrix and genomic DNA was
isolated and purified. 16S ribosomal (r)DNA was amplified by the
polymerase chain reaction (PCR) method using one primer conserved for
members of the domain Bacteria and a second designed specifically for
streptomycetes and related taxa. PCR amplification products were cloned
into phage vector M13 mp19 and the diversity of 16S rDNA genes was
determined by sequence analysis and oligonucleotide probing of the
resultant clone library. Comparison of partial 16S rDNA sequences with
published sequences revealed that few sequences originated from
streptomycetes. The majority of sequences belonged to members of the
alpha subclass of Proteobacteria. Other clones were related to
planctomycetes, actinomycetes, or represented novel lines of descent.
Bacteria that are customarily isolated from soil of pH 4-7 such as
thiobacilli, bacilli, spore- and nonsporeforming actinomycetes, and
pseudomonads are represented in the clone library in small numbers or
were not detected at all. Parameters influencing the recovery,
amplification, quantification, and interpretation of genetic information
from natural sites are discussed.
