Background: F1 hybrids acting as a bridgehead for producing later generation hybrids can have evolutionary significance through strengthening reproductive isolation or facilitating gene flow between parental species, depending on whether backcrossing can occur. It had been suggested that the Tibetan plant Buddleja wardii was a hybrid species between B. alternifolia and B. crispa based on their sympatric distributions and the morphological characters in last century. Till now however, we still have limited evidence to prove key issues to B. wardii, like if it is of hybrid origin indeed and whether it is currently a true hybrid species already.Results: In the present study, two sympatric populations of these three taxa were examined and compared using four nuclear genes and three chloroplast intergenic spacers, as well as with 10 morphological characters. Our results suggest that at both sites B. × wardii was likely to be a hybrids between B. alternifolia and B. crispa, and moreover, most of the hybrids present were confirmed to be F1s. This was further supported by morphology as no transgressive characters were detected. B. crispa was found to be the maternal parent in one population (BH), while in the second population (TJ), it was difficult to distinguish the hybridization direction due to shared haplotypes of cpDNA between B. alternifolia and B. crispa. Conclusions: These results provide evidence that the natural hybrids between B. alternifolia and B. crispa mainly comprise F1 hybrids, which have subsequently been given the name B. wardii. The F1 hybrids have also contributed to strong reproductive isolation between parental species.