Affiliation:
1. Center for Microbial Ecology, Michigan State University, East Lansing
2. Department of Pediatrics and Communicable Diseases, University of Michigan Medical School, Ann Arbor, Michigan
Abstract
ABSTRACT
Despite considerable interest in studying
Burkholderia cepacia
complex in the environment, we still do not have efficient methods to detect, isolate, and screen large numbers of
B. cepacia
isolates. To better describe the ecology and diversity of
B. cepacia
complex, a colony hybridization assay was developed to detect specifically all species of the complex based on polymorphism of the variable V3 region of the 16S rRNA sequence. The sensitivity of the assay was dramatically enhanced by using a probe consisting of three repeats of a
B. cepacia
complex-specific probe, each separated by a phosphoramidite spacer. In addition, a duplex PCR targeting
B. cepacia
complex-specific
recA
and 16S rRNA sequences was developed to enable a fast and reliable diagnostic assay for members of the complex. When applied to maize rhizosphere samples, colony hybridization results were in good agreement with those of most-probable-number duplex PCR, both indicating a >100-fold fluctuation of abundance between individual plants. Using restriction analysis of
recA
for a total of 285 confirmed isolates of the
B. cepacia
complex, up to seven
B. cepacia
complex species were identified; however, their diversity and abundance were not evenly distributed among individual plants, and several allelic variants were commonly found from the same rhizosphere sample. These results indicate that not only complex communities of
B. cepacia
complex species and closely related strains of the same species may coexist at high population levels but also species composition and abundance may dramatically vary between individual plants.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
95 articles.
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