The Himalayan uplift and evolution of aquatic biodiversity across Asia: Snowtrout (Cyprininae: Schizothorax) as a test case

Many morphologically-recognized species have also diverged substantially across basins, suggesting the potential for cryptic speciation and/or undescribed, relictual biodiversity. More detailed molecular studies are needed to test this hypothesis and thus promote conservation planning at the regional scale. This will require the analysis of populations now geographically isolated across the Himalaya, with cooperation/ collaboration needed among neighboring countries for comprehensive, large-scale studies [98, 99]. This is particularly imperative given extant (as well as predicted) impacts of climate change on the Third Pole in general, and Schi- zothhorax in particular [26]. Supporting information S1 Appendix. Divergence intervals (TIMETREE, MEGA X [30]) for Snowtrout (Cyprini- nae: Schizothorax), as associated with geomorphic and climatic drivers in the Himalaya. Confidence intervals for divergence estimates are provided as parentheticals. (PDF) S1 Table. Schizothorax samples from Nepal (N = 53) by site and species. Site = sampling site (N1-N9); Species = Schizothorax sp. identification; River = basin; Location = collection area within basin (confl = confluence); Lat = Latitude; Long = Longitude; KU-T = Univ. Kansas Tissue number (KU:KUIT:#); KU-V = Univ. Kansas Voucher number (KU:KUIT:#); Haplotype = Univ. Arkansas Sequence number; Accession = GenBank Accession number. Sites depicted topographically in Fig 2A. (PDF) S2 Table. Schizothorax samples from Bhutan (N = 19) by site and species. Site = sampling

site (B1-N9); Species = Schizothorax sp. identification; River = basin; Location = collection area within basin (Chhu = river); Lat = Latitude; Long = Longitude; KU-T = Univ. Kansas Tis- sue number (KU:KUIT:#); KU-V = Univ. Kansas Voucher number (KU:KUIT:#); Haplotype = Univ. Arkansas Sequence number; Accession = GenBank Accession number. Sites depicted topographically in Fig 2B. (PDF) S3 Table. Samples of Schizothorax (N = 132) and outgroup species (N = 8) evaluated. Spe- cies Name = Scientific name (A = Acrossocheilus; Pe = Pethia; Pu = Puntius; T = Tor; N = Neo- lissochilus; S = Schizothorax); Haplotype = Univ. Arkansas sequence identifier [i.e., from GenBank = letters and numbers; Univ. of Arkansas = starting with ’58’ (Univ. of Kansas tissue vouchers referenced in S1 Table)]; Location = Country, geographic region or river basin; Group = Regions; Group abbreviations are as follows: QTP = Qinghai-Tibetan Plateau; SEA = Southeastern Asia; YLTR = Yarlung-Tsangpo River; * = indicates samples that cluster with a group outside their sampling region. (PDF) S1 Fig. Phylogenetic tree derived using Bayesian methodology (BEAST v2.61). Phylogenetic relationships among snowtrout (Schizothorax; Cyprinidae) and outgroup taxa derived from sequence analysis of the cytochrome-b mitochondrial gene (1,140 bp, 140 haplotypes). Individ- uals are coalesced into geographic regions, with abbreviations defined as: E-QTP/ SEA = Eastern Qinghai-Tibetan Plateau/ Southeast Asia, with three subclades indicated by A,

B, and C; YLTR = Yarlung-Tsangpo River, with subclades indicated by West-A, West-B, and East. Numbers at nodes represent Bayesian posterior probabilities. Sample metadata in S3 Table. (PDF) PLOS ONE Himalayan uplift and aquatic evolution PLOS ONE | https://doi.org/10.1371/journal.pone.0289736 October 24, 2023 21 / 27