Barcoding ciliates: a comprehensive study of 75 isolates of the genus Tetrahymena

Abstract

The mitochondrial cytochrome-coxidase subunit 1 (cox1) gene has been proposed as a DNA barcode to identify animal species. To test the applicability of thecox1gene in identifying ciliates, 75 isolates of the genusTetrahymenaand three non-Tetrahymenaciliates that are close relatives ofTetrahymena,Colpidium campylum,Colpidium colpodaandGlaucoma chattoni, were selected. All tetrahymenines of unproblematic species could be identified to the species level using 689 bp of thecox1sequence, with about 11 % interspecific sequence divergence. Intraspecific isolates ofTetrahymena borealis,Tetrahymena lwoffi,Tetrahymena patulaandTetrahymena thermophilacould be identified by theircox1sequences, showing <0.65 % intraspecific sequence divergence. In addition, isolates of these species were clustered together on acox1neighbour-joining (NJ) tree. However, strains identified asTetrahymena pyriformisandTetrahymena tropicalisshowed high intraspecific sequence divergence values of 5.01 and 9.07 %, respectively, and did not cluster together on acox1NJ tree. This may indicate the presence of cryptic species. The mean interspecific sequence divergence ofTetrahymenawas about 11 times greater than the mean intraspecific sequence divergence, and this increased to 58 times when all isolates of species with high intraspecific sequence divergence were excluded. This result is similar to DNA barcoding studies on animals, indicating that congeneric sequence divergences are an order of magnitude greater than conspecific sequence divergences. Our analysis also demonstrated low sequence divergences of <1.0 % between some isolates ofT. pyriformisandTetrahymena setosaon the one hand and some isolates ofTetrahymena furgasoniandT. lwoffion the other, suggesting that the latter species in each pair is a junior synonym of the former. Overall, our study demonstrates the feasibility of using the mitochondrialcox1gene as a taxonomic marker for ‘barcoding’ and identifyingTetrahymenaspecies and some other ciliated protists.