TRYPANOSOMATID PROTOZOA IN PLANTS OF SOUTHEASTERN SPAIN - CHARACTERIZATION BY ANALYSIS OF ISOENZYMES, KINETOPLAST DNA, AND METABOLIC BEHAVIOR

Three flagellates of the family Trypanosomatidae were isolated from ma ngo fruits (Mangifera indica) and from the stems of clover (Trifolium glomeratum) and Amaranth (Amaranthus retroflexus) in southeastern Spai n and were adapted to in vitro culture in monophase media. The parasit es showed an ultrastructural pattern similar to that of other species of the genus Phytomonas. Mango and clover isolates differed from amara nth isolates in ultrastructural terms. The isolates were characterized by isoenzymatic analysis and by kDNA analysis using five different re striction endonucleases. With eight of the nine enzymatic systems, man go and clover isolates were distinguished from those of amaranth. Neve rtheless, with the enzymes malate dehydrogenase and superoxide dismuta se, flagellates isolated from clover were differentiated from those is olated from mango. Electrophoretic and restriction endonuclease analys is of kDNA minicircles showed similar restriction cleavage patterns fo r the isolates from mango and clover, whereas the patterns of the amar anth isolates differed. The results of the present study confirm that the strains isolated from mango and clover constitute a phylogenetical ly closely related group of plant trypanosomatids, which is more dista ntly related to the strain isolated from amaranth. The similarities in the results obtained for isolates from mango and clover foliage, on t he one hand, and those obtained from tomato and cherimoya fruits (stud ied previously), on the other, as well as the geographic proximity of the different plants support the contention that only one strain is in volved, albeit one strain that can parasitize different plants. Furthe rmore, some of the plants appear to act as reservoirs for the parasite s. On the other hand, the metabolism studies using [H-1]-nuclear magne tic resonance spectroscopy did not reveal that the catabolism of Phyto monas in general follows a pattern common to all the species or isolat es. Phytomonas are incapable of completely degrading glucose, excretin g a large part of their carbon skeleton into the medium as fermentativ e metabolites (acetate, ethanol, glycine, glycerol, and succinate).