LAUSR

BACKGROUND AND AIMS Species of the genus Buddleja in Asia are mainly distributed in the Sino-Himalayan region and forms a challenging taxonomic group with extensive hybridization and polyploidization. A phylogenetic approach to unraveling the history of reticulation in this lineage will deepen our understanding of the speciation in biodiversity hotspots. METHODS For this study, we obtained 80 accessions representing all the species in the Asian Buddleja clade, and the ploidy level of each taxon was determined by flow cytometry analyses. Whole plastid genomes, nuclear ribosomal DNA, single nucleotide polymorphisms and a large number of low copy nuclear genes assembled from genome skimming data were used to investigate the reticulate evolutionary history of Asian Buddleja. Complex cytonuclear conflicts were detected through a comparison of plastid and species trees. Gene tree incongruence was further analyzed to detect any reticulate events in the history of this lineage. KEY RESULTS Six hybridization events were detected, which are able to explain the cytonuclear conflict in Asian Buddleja. Furthermore, PhyloNet analysis combining species ploidy data indicated several allopolyploid speciation events. A strongly supported species tree inferred from a large number of low copy nuclear genes, not only corrected some earlier misinterpretations, but also indicated that there are many Asian Buddleja species that have been mistakenly lumped. Divergent time estimation shows two periods of rapid diversification (8-10 Mya and 0-3 Mya) in the Asian Buddleja clade, which may coincide, respectively, with the final uplift of the Hengduan Mountains and Quaternary climate fluctuations. CONCLUSIONS This study presents a well supported phylogenetic backbone for the Asian Buddleja species, elucidates their complex and reticulate evolutionary history, and suggests that tectonic activity, climate fluctuations, polyploidization and hybridization together promoted the diversification of this lineage.