Phtheirospermum
(Orobanchaceae), a hemiparasitic genus of Eastern Asia, is characterized by having long and viscous glandular hairs on stems and leaves. Despite this unifying character, previous phylogenetic analyses indicate thatPhtheirospermumis polyphyletic, withPhtheirospermum japonicumallied with tribe Pedicularideae and members of thePh. tenuisectumcomplex allied with members of tribe Rhinantheae. However, no analyses to date have included broad phylogenetic sampling necessary to test the monophyly ofPhtheirospermumspecies, and to place these species into the existing subfamiliar taxonomic organization of Orobanchaceae. Two other genera of uncertain phylogenetic placement areBrandisiaandPterygiella, also both of Eastern Asia. In this study, broadly sampled phylogenetic analyses of nrITS and plastid DNA revealed hard incongruence between these datasets in the placement ofBrandisia. However, both nrITS and the plastid datasets supported the placement ofPh. japonicumwithin tribe Pedicularideae, and a separate clade consisting of thePh. tenuisectumcomplex and a monophyleticPterygiella. Analyses were largely in agreement thatPterygiella, thePtheirospermumcomplex, andXizangiaform a clade not nested within any of the monophyletic tribes of Orobanchaceae recognized to date.Ph. japonicum, a model species for parasitic plant research, is widely distributed in Eastern Asia. Despite this broad distribution, both nrITS and plastid DNA regions from a wide sampling of this species showed high genetic identity, suggesting that the wide species range is likely due to a recent population expansion. ThePh. tenuisectumcomplex is mainly distributed in the Hengduan Mountains region. Two cryptic species were identified by both phylogenetic analyses and morphological characters. Relationships among species of thePh. tenuisectumcomplex andPterygiellaremain uncertain. Estimated divergence ages of thePh. tenuisectumcomplex corresponding to the last two uplifts of the Qinghai-Tibet Plateau at around 8.0-7.0 Mya and 3.6-1.5 Mya indicated that the development of a hot-dry valley climate during these uplifts may have driven species diversification in thePh. tenuisectumcomplex.