An Integrated Morphological and Molecular Approach to a New Species Description in the Trypanosomatidae: the Case of Leptomonas podlipaevi n. sp., a Parasite of Boisea rubrolineata (Hemiptera: Rhopalidae)

Abstract

Leptomonas podlipaevi n. sp., a new trypanosomatid species, is described herein based on light microscopic, ultrastructural, and molecular phylogenetic data. The organism is pleomorphic both in host and culture, with two predominant forms-a typical promastigote with a long flagellum and a shorter promastigote with a small or barely extending flagellum. Several spliced leader RNA repeat sequences obtained from the original cultures and the clonal lines representing two types of cells were all nearly identical. These sequences formed a tight cluster in the neighbor-joining tree well separated from other trypanosomatid species. Glyceraldehyde phosphate dehydrogenase gene sequences were determined for L. podlipaevi and 10 previously described trypanosomatid species. Molecular phylogenetic analysis has demonstrated that the new species is most closely related to Leptomonas seymouri and Leptomonas pyrrhocoris. The analysis has also highlighted the polyphyly of the genus Leptomonas.

Selective recovery of the cultivation-prone components from mixed trypanosomatid infections: a case of several novel species isolated from Neotropical Heteroptera

“…Besides the maximally fully represented 'SE' clade, the analyses included other known major clades of the Trypanosomatidae, each represented by a few relatively slowly evolving members (Hollar et al, 1998;Lukeš et al, 1997;Merzlyak et al, 2001). The overall tree topologies (Figs 4 and 5) were similar to those in previous phylogenetic analyses (Yurchenko et al, 2006a(Yurchenko et al, , b, 2008 and are not discussed herein. It is only important to mention that, although a single gene analysis allows wellsupported clades of trypanosomatids to be revealed, it fails to resolve all deep-branching relationships, which remain a task for the future.…”

“…Cells were processed for light, transmission and scanning electron microscopy as described elsewhere (Yurchenko et al, 2006a(Yurchenko et al, , b, 2008.…”

“…DNA from axenically grown cells was isolated using 10 ml cultures and standard procedures. (thereafter referred to as gGAPDH) was done using the oligonucleotide primers M200, 59-ATGGCTCC(G/A/C)(G/A/C)TCAA(G/A) GT(A/T)GG(A/C)AT-39 and M201, 59-TA(G/T)CCCCACTC(G/A) TT(G/A)TC(G/A)TACCA-39, but otherwise followed procedures described previously (Yurchenko et al, 2006a). Amplification of small-subunit rRNA (SSU rRNA) genes was performed as described previously (Yurchenko et al, 2006b).…”

“…Amplification of small-subunit rRNA (SSU rRNA) genes was performed as described previously (Yurchenko et al, 2006b). Cloning and sequencing procedures used have also been described previously (Yurchenko et al, 2006a).…”

“…Asterisks denote the nodes that were recovered in less than 50 % of replicates. Clade designations (LE, L2, C, P and H) follow our previous nomenclature (Yurchenko et al, 2006a). Bar, 0.05 substitutions per site.…”