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Applied and Environmental Microbiology, January 2001, p. 65-74, Vol. 67, No. 1
Microbial Ecology, UMR-CNRS
5557,1 and INRA,2
Université Claude Bernard-Lyon I, F-69622 Villeurbanne cedex,
France
Received 22 May 2000/Accepted 16 October 2000
Ecology and biodiversity studies of Agrobacterium spp.
require tools such as selective media and DNA probes. Tellurite was tested as a selective agent and a supplement of previously described media for agrobacteria. The known biodiversity within the genus was
taken into account when the selectivity of
K2TeO3 was analyzed and its potential for
isolating Agrobacterium spp. directly from soil was
evaluated. A K2TeO3 concentration of 60 ppm was
found to favor the growth of agrobacteria and restrict the development of other bacteria. Morphotypic analyses were used to define
agrobacterial colony types, which were readily distinguished from other
colonies. The typical agrobacterial morphotype allowed direct
determination of the densities of agrobacterial populations from
various environments on K2TeO3-amended medium.
The bona fide agrobacterium colonies growing on media amended with
K2TeO3 were confirmed to be
Agrobacterium colonies by using 16S ribosomal DNA (rDNA)
probes. Specific 16S rDNA probes were designed for
Agrobacterium biovar 1 and related species
(Agrobacterium rubi and Agrobacterium fici) and
for Agrobacterium biovar 2. Specific pathogenic probes from
different Ti plasmid regions were used to determine the pathogenic
status of agrobacterial colonies. Various morphotype colonies from bulk
soil suspensions were characterized by colony blot hybridization with
16S rDNA and pathogenic probes. All the Agrobacterium-like
colonies obtained from soil suspensions on amended media were found to
be bona fide agrobacteria. Direct colony counting of agrobacterial
populations could be done. We found 103 to 104
agrobacteria · g of dry soil
0099-2240/01/$04.00+0 DOI: 10.1128/AEM.67.1.65-74.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Novel Tellurite-Amended Media and Specific
Chromosomal and Ti Plasmid Probes for Direct Analysis of Soil
Populations of Agrobacterium Biovars 1 and 2
1 in a silt loam bulk
soil cultivated with maize. All of the strains isolated were
nonpathogenic bona fide Agrobacterium biovar 1 strains.
*
Corresponding author. Mailing address: UMR-CNRS
5557 Ecologie Microbienne, Université Claude Bernard Lyon I,
Bât 741, 4ème étage, 43 Bd du 11 Novembre 1918, F-69622 Villeurbanne cedex, France. Phone: 33472448289. Fax:
33472431223. E-mail: nesme{at}bomserv.univ-lyon1.fr.
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