Survival of κ-carrageenan-encapsulated and unencapsulated Pseudomonas aeruginosa UG2Lr cells in forest soil monitored by polymerase chain reaction and spread plating |
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Authors: | Kamtin Leung Michael B. Cassidy Steve B. Holmes Hung Lee Jack T. Trevors |
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Affiliation: | Department of Environmental Biology, University of Guelph, Guelph, Ontario, Canada N1G 2W1;Forest Pest Management Institute, Sault-Ste. Marie, Ontario, Canada P6A 5M7 |
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Abstract: | Abstract A method was developed for direct extraction, purification and amplification of DNA from forest soil. Eighty-two % of the DNA in Pseudomonas aeruginosa UG2Lr introduced into soil was recovered. The detection limit for the strain was approximately 800 cfu g−1 of dry soil based on the polymerase chain reaction (PCR). Survival of κ-carrageenan-encapsulated and unencapsulated UG2Lr was monitored by antibiotic selective and bioluminescence-based nonselective plating and PCR-amplification of a tnsA fragment. After freeze-thaw treatment of soil samples, the unencapsulated UG2Lr declined from an initial population density of 1 × 109 cfu g−1 of dry soil to below the detection threshold of both selective (14 cfu g−1 of dry soil) and nonselective (1 × 103 cfu g−1 of dry soil) plating. However, presence of nonculturable UG2Lr cells in the soil was revealed by PCR and resuscitation of the bacteria. Population density of the encapsulated UG2Lr increased from 2.7 × 106 to 2.9 × 108 cfu g−1 of dry soil after a 3-week incubation at 22°C and declined to 6.3 × 106 cfu g−1 of dry soil after the freeze-thaw treatment. |
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Keywords: | Forest soil luxAB-lacZY reporter genes Polymerase chain reaction Pseudomonas aeruginosa Soil DNA |
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