IN-VITRO CULTURE OF PHYTOMONAS SP ISOLATED FROM EUPHORBIA-CHARACIAS -METABOLIC STUDIES BY H-1-NMR

We describe the in vitro culture of Phytomonas species isolated from E uphorbia characias. The best choice between tested media was SDM-79, i n which promastigotes, after 6 days of culture, reached cell densities as high as 4 x 10(7) cells/ml. Cells growing in LIT or MTL medium sho wed longer division times and lower cell densities. We succeeded in ob taining Phytomonas sp. amastigote and spheromastigote forms in modifie d GRACE's medium, yielding transformation rates of up to 70%. Electron microscopy studies were performed in order to characterize the ultras tructural features of these forms obtained in vitro. On the other hand , metabolic studies based on qualitative (nuclear magnetic resonance s pectroscopy) and quantitative metabolic methods (enzymatic assays) sho wed that promastigote forms secreted mainly ethanol, acetate, glycine, glycerol, piruvate and succinate in SDM-79 medium, whereas the major metabolites found after transformation in modified Grace's medium were ethanol, acetate, glycine, piruvate and smaller amounts of glycerol.