In Vitro Culture of Phytomonas Sp. Isolated from Euphorbia characias. Metabolic Studies by 1H NMR |
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Authors: | MANUEL SÁ NCHEZ-MORENO,CARMEN FERNANDEZ-BECERRA,EMILIO ENTRALA,FRED R. OPPERDOES,,MICHEL DOLLET,ANTONIO OSUNA |
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Affiliation: | Instituto de Biotecnologia, Grupo de Bioquímica y Parasitologia Molecular, Facultad de Ciencias, Universidad de Granada, E-18071 Granada, Spain;Research Unit for Tropical Diseases. International Institute of Cellular and Molecular Pathology, B-1200 Brussels, Belgium;IRHO, Laboratorie de Phytovirologie des Regions Chaudes, CIRAD-ORSTOM, BP5035, 34032 Montpellier cedex 1, France |
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Abstract: | ABSTRACT. 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 × 107 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. |
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Keywords: | Carbohydrate metabolism nuclear magnetic resonance spectroscopy Phytomonas sp. in vitro culture |
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