Burkholderia, a Genus Rich in Plant-Associated Nitrogen Fixers with Wide Environmental and Geographic Distribution

doi:10.1128/AEM.67.6.2790-2798.2001

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Applied and Environmental Microbiology, June 2001, p. 2790-2798, Vol. 67, No. 6
0099-2240/01/$04.00+0 DOI: 10.1128/AEM.67.6.2790-2798.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Burkholderia, a Genus Rich in Plant-Associated Nitrogen Fixers with Wide Environmental and Geographic Distribution

Paulina Estrada-De Los Santos, Rocío Bustillos-Cristales, and Jesús Caballero-Mellado^*

P. de Ecología Molecular y Microbiana, Centro de Investigación sobre Fijación de Nitrógeno, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, Mexico

Received 27 November 2000/Accepted 12 March 2001

The genus Burkholderia comprises 19 species, including Burkholderia vietnamiensis which is the only known N₂-fixing speciesof this bacterial genus. The first isolates of B. vietnamiensiswere recovered from the rhizosphere of rice plants grown in aphytotron, but its existence in natural environments and its geographicdistribution were not reported. In the present study, most N₂-fixingisolates recovered from the environment of field-grown maize andcoffee plants cultivated in widely separated regions of Mexicowere phenotypically identified as B. cepacia using the API 20NEsystem. Nevertheless, a number of these isolates recovered frominside of maize roots, as well as from the rhizosphere and rhizoplaneof maize and coffee plants, showed similar or identical featuresto those of B. vietnamiensis TVV75^T. These features include nitrogenase activity with 10 differentcarbon sources, identical or very similar nifHDK hybridizationpatterns, very similar protein electrophoregrams, identical amplified16S rDNA restriction (ARDRA) profiles, and levels of DNA-DNA reassociationhigher than 70% with total DNA from strain TVV75^T. Although the ability to fix N₂ is not reported to be a commonfeature among the known species of the genus Burkholderia, theresults obtained show that many diazotrophic Burkholderia isolatesanalyzed showed phenotypic and genotypic features different fromthose of the known N₂-fixing species B. vietnamiensis as wellas from those of B. kururiensis, a bacterium identified in thepresent study as a diazotrophic species. DNA-DNA reassociationassays confirmed the existence of N₂-fixing Burkholderia speciesdifferent from B. vietnamiensis. In addition, this study showsthe wide geographic distribution and substantial capability ofN₂-fixing Burkholderia spp. for colonizing diverse host plantsin distantly separatedenvironments.

^* Corresponding author. Mailing address: Centro de Investigación sobre Fijación de Nitrógeno, UNAM, Ap. Postal No. 565-A,Cuernavaca, Morelos, Mexico. Fax: (73) 17-55-81. E-mail: jesuscab{at}cifn.unam.mx.

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