Population structure and evolutionary relationships among desert pupfishes. (a) Principal component analysis of desert pupfishes revealing substantial population structure among species and populations. (b) Species tree estimated by SVDquartets; nonparametric bootstrap support values indicate the percentage of bootstrap replicates supporting monophyly for each clade. (c) Maximum-likelihood tree of all 30 samples estimated by IQ-TREE. Two internal branches were collapsed due to low support (ultrafast bootstrap < 95%, SH-aLRT < 80%); all other branch support was unequivocal (UFboot = 100%, SH-aLRT = 100%). (d) Ancestry proportions across individuals in Death Valley NP, Ash Meadows NWR and outgroup desert pupfishes (C. albivelis, C. eremus, C. fontinalis, C. macularius, C. radiosus) estimated from a LD-pruned SNP dataset in ADMIXTURE with k = 4. Colours in PCA, trees and ADMIXTURE designate individuals from different species or populations and correspond to the shared figure legend in (a). (Online version in colour.)