Determination of phospholipid ester-linked fatty acids and poly β-hydroxybutyrate for the estimation of bacterial biomass and activity in the rhizosphere of the rape plant Brassica napus (L.)

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Abstract
Examination of seven strains of gram-negative bacteria isolated from the rhizosphere of the rape plant showed profiles of fatty acids, ester-linked to their phospholipids that were distinctly different from sterile roots. The bacteria were enriched in short and branched saturated, cyclopropane and monoenoic fatty acids in which the unsaturation was formed by the anaerobic desaturase pathway when growth was on a medium simulating root exudates. This suite of fatty acids was significantly increased in sand and in roots of rape plants grown from seeds in sands inoculated with these organisms at the start of the experiments. Some of the bacteria formed the endogenous storage polymer poly .beta.-hydroxybutyrate in monocultures. Poly .beta.-hydroxybutyrate was formed by the bacteria in the inoculated sand in large amounts but not by the bacteria that were recovered from the roots. This suggests that bacteria associated with the roots were in balanced growth, whereas the bacteria in the sand were lacking essential nutrients and showed an unbalanced growth response. These chemical analyses provided estimates of the bacterial biomass (from the sum of bacterial ester-linked phospholipid fatty acids) and the metabolic status (poly .beta.-hydroxybutyrate to phospholipid fatty acid ratio) in this experiment. With further definition of lipid biomarkers of soil microorganisms these experimental techniques may be extended to the rhizosphere.