Classification of trypanosomatids from insects and plants by the UP-PCR (Universally Primed PCR) technique and cross dot blot hybridization of PCR products

For the first time the UP-PCR technique with its hybridization assay was applied to trypanosomatids isolated from insects and plants. 13 isolates of trypanosomatids from insects (eight isolates from Russia, one from Czech Republic and four reference cultures) and three plant isolates were analyzed....

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Bibliographic Details
Published in:European journal of protistology 1999-10, Vol.35 (3), p.319-326
Main Authors: Bulat, Sergei A., Mokrousov, Igor V., Podlipaev, Sergei A.
Format: Article
Language:English
Subjects:
Classification
Crithidia
Cross dot blot hybridization
Czech Republic
Herpetomonas
Leptomonas
PCR
Russia
Trypanosomatidae
Wallaceina
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Summary:For the first time the UP-PCR technique with its hybridization assay was applied to trypanosomatids isolated from insects and plants. 13 isolates of trypanosomatids from insects (eight isolates from Russia, one from Czech Republic and four reference cultures) and three plant isolates were analyzed. By the cross dot blot hybridization of UP-PCR products it was demonstrated that most trypanosomatids except for Wallaceina branch and two species of Leptomonas genera are quite distant from each other and the degree of relationship cannot be resolved by PCR with random primers. The methods used allowed segregation of all trypanosomatids tested into 12 separate genospecies (natural groups) with very different genomic structure. Wallaceina together with two others isolates and two Leptomonas formed separate groups. Therefore, separate taxon status for Wallaceina and two Leptomonas species as well as heterogeneity of Leptomonas, Herpetomonas and Crithidia genera are proposed. The low host specificity of insect trypanosomatids has been demonstrated — two isolates of parasites from different insect orders are more similar than isolates from the same insect species. Isolation of occasional parasite or mixed parasite populations in laboratory culture is discussed. It may be proposed from the data obtained that the genera existing now do not reflect the real biodiversity of trypanosomatids and the current generic systematics of trypanosomatids have to be reviewed. Additional experiments involving a large number of new isolates are necessary to resolve this problem.
ISSN:0932-4739
1618-0429
DOI:10.1016/S0932-4739(99)80010-8