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In Vitro Culture of Phytomonas Sp. Isolated from Euphorbia characias. Metabolic Studies by 1 H NMR
Author(s) -
SÁNCHEZMORENO MANUEL,
FERNANDEZBECERRA CARMEN,
ENTRALA EMILIO,
OPPERDOES FRED R.,
DOLLET MICHEL,
OSUNA ANTONIO
Publication year - 1995
Publication title -
journal of eukaryotic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 1066-5234
DOI - 10.1111/j.1550-7408.1995.tb01586.x
Subject(s) - biology , glycerol , glycine , in vitro , cytochemistry , biochemistry , amastigote , ultrastructure , cell culture , botany , enzyme , parasite hosting , amino acid , genetics , world wide web , computer science , leishmania
. We describe the in vitro culture of Phytomonas species isolated from Euphorbia characias . The best choice between tested media was SDM‐79, in which promastigotes, after 6 days of culture, reached cell densities as high as 4 × 10 7 cells/ml. Cells growing in LIT or MTL medium showed longer division times and lower cell densities. We succeeded in obtaining Phytomonas sp. amastigote and spheromastigote forms in modified GRACE's medium, yielding transformation rates of up to 70%. Electron microscopy studies were performed in order to characterize the ultrastructural features of these forms obtained in vitro. On the other hand, metabolic studies based on qualitative (nuclear magnetic resonance spectroscopy) and quantitative metabolic methods (enzymatic assays) showed 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.