Multiplexed shotgun genotyping (MSG) data resolve phylogenetic relationships within and among archipelagos in Macaronesian Tolpis

Premise: Plants endemic to oceanic archipelagos are suitable for studying evolution, being isolated on substrates of different ages. Evolution has been recent, rendering traditionally employed sequences insufficiently variable for resolving relationships. This study includes sampling in the genus Tolpis (Asteraceae) from the Azores, Madeira, and Cape Verde, and expands upon an earlier study demonstrating the efficacy of multiplexed shotgun genotyping (MSG) for resolving relationships in Canarian Tolpis. Methods: Genomic libraries for 90 accessions of Tolpis and two from the outgroup were generated for genotyping individuals using MSG. Loci were de novo assembled with iPyrad, which clusters MSG loci within and between samples. A maximum likelihood phylogeny was generated with RAxML. Ancestral area reconstruction was inferred using R package BioGeoBEARS. Results: MSG data recovered a highly resolved phylogeny from population to inter-archipelago levels. Ancestral area reconstruction provided biogeographic hypotheses for the radiation of Macaronesian Tolpis. Conclusions: Four major clades were resolved. The Madeiran endemic T. macrorhiza is sister to other Tolpis. Species from the Canaries, Cape Verdes, and the continent are sister to T. succulenta from Madeira, which has a sister subclade of Azorean populations composed of T. succulenta and T. azorica. Population-level resolution suggests unrecognized taxa on several archipelagos. Ancestral reconstruction suggests initial dispersal from the continent to Madeira, with dispersal to the Azores, then dispersal from Madeira to the Canary Islands, with both subsequent dispersal to the Cape Verdes and back-dispersal to the continent. Single-island radiations and inter-island dispersal are implicated in divergence in Macaronesian Tolpis.