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Applied and Environmental Microbiology, March 2005, p. 1193-1201, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1193-1201.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Species Abundance and Diversity of Burkholderia cepacia Complex in the Environment

Alban Ramette,1 John J. LiPuma,2 and James M. Tiedje1*

Center for Microbial Ecology, Michigan State University, East Lansing,1 Department of Pediatrics and Communicable Diseases, University of Michigan Medical School, Ann Arbor, Michigan2

Received 23 July 2004/ Accepted 15 October 2004

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.

* Corresponding author. Mailing address: Center for Microbial Ecology, 540 Plant and Soil Sciences Building, Michigan State University, East Lansing, MI 48824-1325. Phone: (517) 355-0271, ext. 285. Fax: (517) 353-2917. E-mail: tiedjej{at}msu.edu.

Applied and Environmental Microbiology, March 2005, p. 1193-1201, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1193-1201.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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