New media (S1 and S2) were formulated that provide a high degree of selectivity and detection of fluorescent pseudomonads on initial plating. The selectivity of the S-type media was based on a detergent, sodium lauroyl sarcosine, and an antibiotic, trimethoprim. A total of five soils from different geographical locations and one sewage sludge sample were examined. On S1 medium, isolates from two soils with low fluorescent pseudomonad populations exhibited a high frequency of arginine dihydrolase (78%) and oxidase-positive (95%) phenotypes, but no fermentative isolates were recovered. Medium S2 was more defined and selective than S1, but lower numbers of fluorescent pseudomonads were recovered on S2. In soils in which fluorescent pseudomonads represent a small proportion of the total population, S1 medium consistently recovered high percentages of fluorescent phenotypes (82.5%).
TB-T medium provides a high degree of selectivity for and detection of Pseudomonas cepacia biotypes upon initial plating from soil. TB-T medium consists of a basal medium with glucose as the sole carbon source and asparagine as the sole nitrogen source. The selectivity of TB-T medium is based on the combination of trypan blue (TB) and tetracycline (T) (pH 5.5). On TB-T medium, 216 of 300 isolates (72%) from five different soil types were identified as P. cepacia. The remaining 28% were facultative organisms that could be separated readily from P. cepacia by anaerobic glucose fermentation and by their inability to grow at 41°C. Molds were controlled on low soil dilutions by adding crystal violet, nystatin, or both. Elimination of either ingredient or elevation of the pH to 7.5 resulted in a pronounced loss of selectivity. The efficiency of recovery varied considerably among P. cepacia strains but was high enough for some strains (76 to 86%) to permit quantitative studies. TB-T medium combines a defined formulation with high selectivity and allows recovery of P. cepacia biotypes from low soil dilutions (10' to 103).
During the 1983 field season, the rhizobacteria (including organisms from rhizosphere soil and the root rhizoplane) of cotton plants at one location in Mississippi were inventoried at different plant growth stages. Isolates (1,000) were identified to the genus level and characterized for repression of Pythium ultimum and Rhizoctonia solani. Cotton seedlings were initially colonized by bacteria of many different genera, and populations quickly reached 10 8 CFU/g of root tissue. As the season progressed, the bacterial populations declined as root mass increased and the roots became more woodlike in consistency. Fluorescent pseudomonads were the most numerous gram-negative rhizobacterial isolates of those that were randomly collected and identified, and they provided the largest number of isolates with fungal repressive activity. Several other gram-negative bacterial genera were recovered throughout the growing season, and some gram-positive bacteria were also isolated routinely, but at lower numbers. There was no correlation between the proportion of rhizobacterial isolates that possessed fungal repressive activity and the plant growth stage from which the isolates were obtained. Approximately twice as many bacterial isolates demonstrated fungal repression in the agar assay compared with the inplanta assay, and isolates were found more frequently with fungal repressive activity against P. ultimum than against R. solai.
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