Diversity and Phylogeny of Trichonympha, a Parabasalian Symbiont of Termites

Trichonympha is a Parabasalian symbiont of lower termites. They are characterized by their spindle-shape, length ranging from 75-150 micrometers, longitudinal flagella, as well as their true rostral tube composed of two plates of parabasal fibers. Ealy analysis through microscopy has revealed that the same species of Trichonympha could be found across multiple species of termites. However, with recent phylogenetic analysis of the 18S region, it has been found that the species is actually genetically distinct and that Trichonympha have coevolved with their termite hosts. In this study, Trichonympha cells from Reticulitermes tibialis, R. flavipes, and R. lucifugus were isolated. DNA from the single cell samples were purified and amplified through a series of polymerase chain reactions (PCR) and gel electrophoresis. The amplified DNA was extracted from the gel and cloning was performed using competent E. coli. The colonies that formed from each sample were collected and those that amplified after a PCR reaction were sent to the ASU Genomics Core for sanger sequencing. The sequences were assembled, cleaned, and aligned and the maximum likelihood and Bayesian phylogenetic trees were constructed. It was found that there is evidence of multiple species of Trichonympha in R. flavipes and that there were multiple species found in R. lucifugus. However, the Trichonympha sequences from R. lucifugus branch in different clades which indicates that they are more distant species than those of R. flavipes. Additionally, Trichonympha cells from R. tibialis branched in the same clade as those from R. flavipes which provides evidence that the same Trichonympha species can be found in multiples species of Reticulitermes. With these pieces of evidence, we can see multiple patterns of diversity of Trichonympha in their termite hosts.