Utility of low-copy nuclear markers in phylogenetic reconstruction of Hypericum L. (Hypericaceae)

Abstract

Primers and sequence variation for two low-copy nuclear genes (LCG) not previously used for phylogenetic inference in the genus Hypericum, PHYC and EMB2765, are presented here in comparison with the fast-evolving nuclear intergenic spacer ITS. Substitution rates in the LCG markers were half those reported in ITS for Hypericum, which might help avoid the problems caused by substitution saturation and difficulties to establish homologies that afflict the latter marker. We included representatives of all major clades within Hypericum and found that levels of phylogenetic resolution, clade support values and internal character consistency were similar to, or even higher than, those of ITS-based phylogenies. The presence of at least two copies in EMB2765 in Hypericum imposed a methodological challenge that was circumvented by the design of an effective clade-specific primer. Both EMB2765 and, especially, PHYC appear to be good alternatives to the ITS marker, confirming the main phylogenetic relationships found in previous studies, but with improved resolution and support values for some basal relationships. © 2014 Springer-Verlag Wien.