PCR amplification of the spliced leader gene for the diagnosis of trypanosomatid parasites of plants and insects in methanol-fixed smears |
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Authors: | Myrna G. Serrano Marta Campaner Gregory A. Buck Marta M.G. Teixeira Erney Plessmann Camargo |
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Affiliation: | 1. Institute of Parasitology, Biology Centre, Czech Academy of Sciences, Branišovská 31, 37005 České Budějovice, Czech Republic;2. Faculty of Science, University of South Bohemia, Branišovská 31, 37005 České Budějovice, Czech Republic |
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Abstract: | A PCR-based method was adapted for the amplification of DNA from methanol-fixed smears of insects and plants parasitized by trypanosomatids. The PCR target was the multicopy spliced leader (SL) gene. Amplicons were hybridized with an oligonucleotide probe (SL3') specific for Phytomonas. The method has the advantage of dispensing with the cultivation of parasites, many of which are very fastidious or non-cultivable. The technique was applied to archival glass slides and to newly collected material. It proved to specific for Phytomonas spp., enabling their detection in plants and insects. Sequence comparison of the amplicons obtained revealed the existence of different strains/species of Phytomonas circulating among diseased palsms and fruit. |
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Keywords: | Palm disease Plant trypanosomatid Insect trypanosomatid Archival DNA |
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