The role of birds and insects in pollination shifts of Scrophularia (Scrophulariaceae)

[Display omitted] •DNA sequences analyses suggest that Scrophularia diverged in the Miocene.•Wasp pollination is the ancestral pollination system in Scrophularia.•The mixed vertebrate-insect pollination system was acquired independently four times.•There is no evidence for an ancient role of hummingbirds in the evolution of European Scrophularia.•A continental wasp-pollinated ancestor is suggested for the Macaronesian S. calliantha. The mixed vertebrate-insect pollination system is rare in Holarctic plants. Phylogenetic relationships of 116 Scrophularia taxa were investigated based on two plastid (ndhF and trnL-trnF) and one nuclear (ITS) DNA regions. A wider time-calibrated analysis of ndhF sequences of the Lamiales revealed that Scrophularia diverged as early as in the Miocene (