Comparative analysis of antibiotic resistance, immunofluorescent colony staining, and a transgenic marker (Bioluminescence) for monitoring the environmental fate of a rhizobacterium

• Published in: Applied and Environmental Microbiology Vol. 63; no. 4; pp. 1617 - 1622
• Main Authors: Mahaffee, W.F. (USDA, ARS, Horticulture Crops Research Laboratory, Corvallis, OR.), Bauske, E.M, Vuurde, J.W.L. van, Wolf, J.M. van der, Brink, M. van den, Kloepper, J.W
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01-04-1997
• Subjects: Agronomy. Soil science and plant productions; Anti-Bacterial Agents - pharmacology; BACTERIA; Bacterial Typing Techniques; Biochemistry and biology; Biological and medical sciences; Biotechnology; Chemical, physicochemical, biochemical and biological properties; CUCUMIS SATIVUS; Drug Resistance, Microbial; Environment and pollution; Environmental Microbiology; Fluorescent Antibody Technique; Fundamental and applied biological sciences. Psychology; Genetically engineered organisms behavior (microorganisms, plants, animals); Industrial applications and implications. Economical aspects; Instituut voor Plantenziektenkundig Onderzoek; Luminescence; Microbiology; Physics, chemistry, biochemistry and biology of agricultural and forest soils; Pseudomonas fluorescens; Research Institute for Plant Protection; Rhizobiaceae - classification; Rhizobiaceae - drug effects; RHIZOSPHERE; RIZOSFERA; Soil science; Pseudomonadales; Recombinant microorganism; Bioluminescence; Cucurbitaceae; Plant growth-promoting rhizobacteria; Pseudomonas fluorescens; Cucumis sativus; Field experiment; Resistance; Rhizosphere; Antibiotic; Rhizobacteria; Vegetable crop; Dicotyledones; Angiospermae; Bacteria; Pseudomonadaceae; Spermatophyta; Population dynamics; Immunofluorescence; Comparative study; Monitoring
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/aem.63.4.1617-1622.1997

Field releases of the wild-type plant growth-promoting rhizobacterium Pseudomonas fluorescens 89B-27, its bioluminescent derivative GEM-8 (89B-27::Tn4431), and a spontaneous rifampin-resistant variant (R34) were established on cucumber in 1994 and 1995 to examine the efficiency of these marker systems for estimating the wild-type population. Seed and root samples were taken 0, 7, 14, 21 or 28, 35 or 42, and 70 days after planting in each year and processed for enumeration by spiral plating or immunofluorescent colony staining (IFC). In both years, the populations of 89B-27, R34, and GEM-8, as measured by IFC, were not significantly different (P 0.05) from each other at each sampling time. However, the populations of R34 and GEM-8, as measured by spiral plating and differentiation based on their respective phenotypes, were significantly lower (P 0.05) than the wild-type populations and their IFC-determined populations. These data indicate that traditional marker systems may underestimate populations and hence the survival and colonization of genetically marked bacteria