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Application of a high-performance liquid chromatography-based muramic acid assay with precolumn fluorescence derivatization to quantification of root-associated bacteria was studied both in pure cultures and in the rhizosphere of axenicFestuca rubra seedlings. Quantities of muramic acid from acid-hydrolyzed cells ofFrankia strains,Streptomyces griseoviridis, Enterobacter agglomerans, Klebsiella pneumoniae, Pseudomonas sp., andBacillus polymyxa were mostly proportional to the respective cell protein and carbon quantities, but in some strains, culture age and particularly sporulation affected these ratios considerably. The muramic acid/cell protein ratio was generally 2 to 4 times higher in strains of the two actinomycete genera,Frankia andStreptomyces , than in the rest of the strains. Quantification ofFrankia strains,S. griseoviridis, E. agglomerans , andPseudomonas sp. was also attempted from the rhizosphere ofF. rubra seedlings which had been inoculated with pure cultured bacteria and incubated briefly. It was possible to quantifyFrankia cells by use of the muramic acid assay from both the root and the growth medium, whereas cells of the rest of the bacterial genera could only be detected in the medium. The detection limit for muramic acid was about 10 ng/ml hydrolysis volume, and from theFestuca rhizosphere, 28 to 63% of the muramic acid in theFrankia inoculum was recovered.

applied and environmental microbiology

Quantification of Frankia Strains and Other Root-Associated Bacteria in Pure Cultures and in the Rhizosphere of Axenic Seedlings by High-Performance Liquid Chromatography-Based Muramic Acid Assay