The influence of naturally occurring phenolic acids on phosphate uptake by barley (Hordeum vulgare L. cv. Karlsberg) roots was examined using '3P-labeled phosphate. Without exception, all compounds tested, namely, benzoic, 2-hydroxybenzoic, 4-hydroxybenzoic, 3,4-dihydroxybenzoic, 3,4,5-trihydroxybenzoic, 4-hydroxy-3-methoxybenzoic, 4-hydroxy-3,5-dimethoxybenzoic, cinnamic, 2-hydroxycinnamic, 4-hydroxycinnamic, 3,4-dihydroxycinnamic, 4-hydroxy-3-methoxycinnamic, and 4-hydroxy-3,5-dimethoxycinnamic acids, inhibited uptake. (24,25).It has been demonstrated that in both higher plants (8) (12), and ferns (7). Through decomposition of plant material, rain leaching from leaves and bark. and exudation from roots a wide variety of phenolic acids may be introduced into the soil solution. Whitehead (23) sampled a variety of different soil types and reported the presence of 4-hydroxybenzoic, 4-hydroxy-3-methoxybenzoic, 4-hydroxycinnamic, and 4-hydroxy-3-methoxycinnamic acids as the free acids at soil solution concentrations equivalent to 5 X IO's M.The present study examines the influence of these naturally occurring phenolic acids upon inorganic phosphate uptake by barley roots. Concentrations employed were in the range reported by Whitehead (23). In the main, experiments were of short duration ranging from several minutes to 3 hr.
MATERIALS AND METHODSBarley seeds (Hordeum vulgare L. cv. Karlsberg) were germinated and prepared for treatment according to the methods described by Epstein and Hagen (5). All experiments were performed with 4-day-old excised roots, approximately 3 cm long. After excision the roots were gently mixed in a large volume of distilled water, filtered through two layers of cheesecloth, and spun by hand in a cheesecloth bag to remove excess water.Root samples weighing 2 g were placed in 150-ml beakers containing 50 ml of solution, buffered at pH 7, containing 5 X I0-3 M Pi, 5 X 10-4 M CaCl2, and 0.05 pc of 3P-labeled phosphate. All experiments were performed at room temperature (23 C), and solutions were aerated continuously during the uptake period. The various phenolic acids which were added to the treatment solutions were dissolved in 1 ml of 95% ethanol. In most experiments, the final concentrations of phenolic acids experienced by the root tissue were 5 x 10' M, slightly in excess of the values reported by Whitehead (23) fer soil solution. In the preliminary kinetic experiments involving vanillic acid, concentrations ranged from 5 X 10-5 M to 1 X 10' M.The uptake of phosphate by root tissue was determined by the removal of 1-ml aliquots from the incubation medium at prescribed intervals. The radioactivity of each sample was determined in a Packard scintillation spectrometer, and uptake was determined by loss of activity from the solution.Preliminary experiments indicated that the uptake of phosphate, in both control treatments (in the absence of inhibitors) and vanillic acid treatments, had become linear within 5 to 10 min of immersion of the tissue in the phosphate solutions. In the com...