Aim Dioscorea (Dioscoreaceae) is a predominantly pantropical genus (< 600 species) that includes the third most important tropical tuber crop and species of pharmacological value. Fossil records from both the Northern and Southern Hemispheres were used to test hypotheses about the origin of the genus Dioscorea, and to examine potential macroevolutionary processes that led to its current distribution. Location Pantropical distribution.Methods Divergence times were estimated using the most comprehensive phylogeny of the group published to date based on plastid sequences and fossil calibrations, applying a relaxed-clock model approach. Ancestral areas and range shifts were reconstructed using time-stratified likelihood-based models, reflecting past continental connectivity and biogeographical models incorporating the spatial range of fossils.Results Fossil-informed biogeographical analysis supported colonization of the Nearctic by ancient yam lineages from the western Palaearctic and subsequent migration to the South. Most of the pantropical South American, African and Southeast Asian lineages experienced a relatively recent diversification in the Oligocene-Miocene. Long-distance dispersals were inferred for the colonizations of the New World, Africa and Madagascar.Main conclusions Dioscorea likely originated between the Late Cretaceous and the Early Eocene in the Laurasian Palaearctic, followed by possible dispersal to South America via the Eocene North Atlantic Land Bridge.

Citation: Hale, H., E. M. Gardner, J. Viruel, L. Pokorny, and M. G. Johnson. 2020. Strategies for reducing per-sample costs in target capture sequencing for phylogenomics and population genomics in plants. Applications in Plant Sciences 8(4): e11337.The reduced cost of high-throughput sequencing and the development of gene sets with wide phylogenetic applicability has led to the rise of sequence capture methods as a plausible platform for both phylogenomics and population genomics in plants. An important consideration in large targeted sequencing projects is the per-sample cost, which can be inflated when using off-the-shelf kits or reagents not purchased in bulk. Here, we discuss methods to reduce per-sample costs in high-throughput targeted sequencing projects. We review the minimal equipment and consumable requirements for targeted sequencing while comparing several alternatives to reduce bulk costs in DNA extraction, library preparation, target enrichment, and sequencing. We consider how each of the workflow alterations may be affected by DNA quality (e.g., fresh vs. herbarium tissue), genome size, and the phylogenetic scale of the project. We provide a cost calculator for researchers considering targeted sequencing to use when designing projects, and identify challenges for future development of low-cost sequencing in non-model plant systems. KEY WORDS enzymatic fragmentation; herbariomics; high-throughput workflow implementation; Hyb-Seq; low-cost sequence capture; pooling and multiplexing strategies. Applications in Plant Sciences 2020 8(4): e11337 Hale et al.-Low-cost Hyb-Seq • 2 of 10

Aim Phylogeography of fruit trees is challenging due to recurrent exchanges between domesticated and wild populations. Here we tested the eastern refugium hypothesis (ERH) for the carob tree, Ceratonia siliqua, which supports its natural and domestication origins in the eastern Mediterranean and a feral origin in the west. Location Mediterranean basin. Taxon Ceratonia siliqua L., Leguminosae. Methods A phylogenetic reconstruction based on two nuclear and one plastid sequences was performed to estimate the divergence time between the carob tree and its sister species, Ceratonia oreothauma. Variation from four plastid regions and 17 nuclear microsatellite loci were used to decipher genetic structure in the carob tree and to test coalescent‐based models by an Approximate Bayesian computation (ABC) approach. We assessed our hypotheses by examining palaeobotanical records and hindcasting the past distribution of the carob tree at Mid‐Holocene, Last Glacial Maximum (LGM) and Last Interglacial (LIG) using species distribution modelling. Results The split between C. oreothauma and C. siliqua was estimated at 6.4 Ma, and a first divergence within C. siliqua at 1.3 Ma. After a continuous presence since the Oligocene, Ceratonia was rarely found in the fossil record during the Pleistocene but present in the western and the eastern Mediterranean. Plastid and nuclear markers, characterized by low allelic richness, revealed a strong west‐east genetic structuring. ABC analyses rejected the ERH. Main conclusions Our study supports a severe population decline during LIG. The strong west–east divergence and the occurrence of four lineages within C. siliqua provided support for a new hypothesis of multiple domestications of the carob tree from native populations throughout the Mediterranean basin.