Detection of Pseudomonas putida B in the rhizosphere by RAPD |
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Authors: | B.M. Hansen,& A. Winding |
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Affiliation: | Department of Marine Ecology and Microbiology, National Environmental Research Institute, Roskilde, Denmark |
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Abstract: | A method was developed for the detection of Pseudomonas putida B MM12 released into the rhizosphere of non-sterile barley, using a Random Amplified Polymorphic DNA (RAPD)-generated probe for hybridization with RAPD products generated from DNA extracted from the rhizosphere. The detection procedure involves extraction of rhizosphere bacteria by sonication, extraction of DNA by boiling, RAPD and Southern hybridization with RAPD products and the selected probe. The level of detection of MM12 was at least 1·9×104 cells g−1 barley root. MM12 was detected in rhizosphere when it constituted as little as 0·5% of the culturable population. |
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