Trypanosomatidae: a spliced-leader-derived probe specific for the genus Phytomonas

Experimental Parasitology
M M TeixeiraE P Camargo

Abstract

We probed DNA from all trypanosomatid genera by slot blot hybridization with an oligonucleotide (SL3') complementary to a sequence of the Phytomonas spliced-leader or mini-exon RNA. The 19-nucleotide probe target site was previously shown to be highly conserved among a limited number of Phytomonas isolates, but diverges in other kinetoplastid genera. Our examination of 84 isolates of various genera of trypanosomatids showed hybridization of this probe exclusively with isolates from plants or insects which could, by morphological, biochemical, and molecular criteria, be considered to belong to the genus Phytomonas. In contrast, no hybridization was observed with flagellates of the genera Blastocrithidia, Crithidia, Endotrypanum, Herpetomonas, Leptomonas, Leishmania, and Trypanosoma. The method detected DNA quantities as low as 50 ng using either radioactive or nonradioactive probes, and was effective with as few as 10(4) intact flagellates. Together, these results suggest that this probe will serve as a convenient marker for taxonomic and epidemiological studies requiring reliable identification of Phytomonas spp. in plants or in putative insect vectors.

Citations

May 5, 2005·Molecular and Biochemical Parasitology·Georgios J PappasAntonio González
Aug 22, 2002·The Journal of Eukaryotic Microbiology·Mara M I GodoiErney P Camargo
May 30, 2001·Molecular and Biochemical Parasitology·J M QuesadaM Sánchez-Moreno
Nov 19, 1997·Molecular and Biochemical Parasitology·L Hollar, D A Maslov

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