Affiliation:
1. Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, Michigan
Abstract
ABSTRACT
An integrative approach was used to obtain pure cultures of previously uncultivated members of the divisions
Acidobacteria
and
Verrucomicrobia
from agricultural soil and from the guts of wood-feeding termites. Some elements of the cultivation procedure included the following: the use of agar media with little or no added nutrients; relatively long periods of incubation (more than 30 days); protection of cells from exogenous peroxides; and inclusion of humic acids or a humic acid analogue (anthraquinone disulfonate) and quorum-signaling compounds (acyl homoserine lactones) in growth media. The bacteria were incubated in the presence of air and in hypoxic (1 to 2% O
2
[vol/vol]) and anoxic atmospheres. Some bacteria were incubated with elevated concentrations of CO
2
(5% [vol/vol]). Significantly more
Acidobacteria
were found on isolation plates that had been incubated with 5% CO
2
. A simple, high-throughput, PCR-based surveillance method (plate wash PCR) was developed. This method greatly facilitated detection and ultimate isolation of target bacteria from as many as 1,000 colonies of nontarget microbes growing on the same agar plates. Results illustrate the power of integrating culture methods with molecular techniques to isolate bacteria from phylogenetic groups underrepresented in culture.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
329 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献