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Calcium dependent protein phosphorylation was studied in corn (Zea mays L.) root tips.  Prior to in vivo protein phosphorylation experiments, the effect of calcium, ethyleneglycol-bis-(beta-aminoethyl ether)-N-N'  -tetraacetic acid (EGTA) and calcium ionophore (A-23187) on phosphorus uptake was studied.  Calcium increased phosphorus uptake, whereas EGTA and A-23187 decreased it.  Consequently, phosphorus concentration in the media was adjusted so as to attain similar uptake in different treatments.  Phosphoproteins were analyzed by two-dimensional gel electrophoresis.  Distinct changes in phosphorylation were observed following altered calcium levels.  Calcium depletion in root tips with EGTA and A-23187 decreased protein phosphorylation.  However, replenishment of calcium following EGTA and ionophore pretreatment enhanced phosphorylation of proteins.  Preloading of the root tips with 32P in the presence of EGTA and A-23187 followed by a ten minute calcium treatment, resulted in increased phosphorylation indicating the involvement of calcium, calcium and calmodulin-dependent kinases.  Calmodulin antagonist W-7 was effective in inhibiting calcium-promoted phosphorylation.  These studies suggest a physiological role for calcium-dependent phosphorylation in calcium-mediated processes in plants.

Calcium-Regulated in Vivo Protein Phosphorylation in Zea mays L. Root Tips