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Deciphering Evolutionary Ancestry in African Tree Frogs: Genus Leptopelis Kyle E

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Deciphering Evolutionary Ancestry in African Tree Frogs: Genus Leptopelis Kyle E Deciphering Evolutionary Ancestry in African Tree Frogs: Genus Leptopelis Kyle E. Jaynes1,2 and Rayna C. Bell1 1Department of Vertebrate Zoology, National Museum of Natural History, Smithsonian Institution 2Department of Biology and Environmental Sciences, Adrian College Morphological Analyses Introduction Phylogeny of 30 Leptopelis Species Biogeographical Regions of Africa The genus Leptopelis is a group of African Tree Frogs that have a a) b) poorly understood taxonomic history, with 53 currently accepted Size species (Frost 2017). These species are closely related and difficult to L_parbocag!! (58mm)! L. parbocagii distinguish by morphological features alone, often times being L_barbouri (43mm)! L. barbouri L_parkeri !! (56mm)! L. parkeri L_argenteu (52mm)! L. argenteus misidentified in the field. Leptopelis palmatus is an endemic species L_natalens (65mm)! L. natalensis L_yaldeni!! (43mm)! L. yaldeni to Príncipe Island, one of four islands in the Gulf of Guinea L_vannutel (46mm)! L. vannutellii L_cf_ragaz!! (50mm)! L. ragazzii archipelago (Jones 1994). Reconstructing its colonization history and L_susanae (53mm)! L. susanae L_gramineu!! (63mm)! L. gramineus Introduction L_fiziensi (38mm)! L. fiziensis subsequent speciation i s key to understanding the biogeography of L_vermicul (85mm)! L. vermiculatus L_kivuensi!! (36mm)! L. kivuensis this archipelago. L_flavomac (70mm)! L. flavomaculatus L. macrotis, L. millsoni, and L. rufus form a species complex that L_bocagii!! (58mm)! L. bocagii L_modestus (41mm)! L. modestus L_palmatus (110mm)! L. palmatus! occurs across West and Central Africa, as well as on Bioko Island, a L_aubriodi! ! (43mm)! L. aubryioides L_notatus (74mm)! L. notatus L_breviros continental land-bridge system. These species have been !! (64mm)! L. brevirostris Leptopelis rufus Leptopelis millsoni L_calcarat (57mm)! L. calcaratus hypothesized to be the closest ancestors of L. palmatus due to their L_boulenge!! (81mm)! L. boulengeri L_ocellatu (58mm)! L. ocellatus Figure 3– Hand sketch of webbing extent in L. millsoni and L. rufus. L_aubryi (54mm)! L. aubryi large size and proximity to Príncipe (Schiotz 1999). In addition, their L_viridis!! (48mm)! L. viridis distribution across continental Africa and Bioko Island make them an L_spiritus (49mm)! L. spiritusnoctis L_macrotis!! (84mm)! L. macrotis! Among other characters, the extent of webbing in feet has ideal study system for investigating speciation across a range of L_millsoni (87mm)! L. millsoni! L_rufus (87mm)! L. rufus! been a consistently distinguishable tool for our specimens. We Astylo_spp! environments. In this project, we investigate questions of (NA) Astylosternus spp. Photo credit: A. Stanbridge 0.03 Leptopelis palmatus have also found that the ratio of disc (DSC) to Tympanum phylogeography and ancestral history in order to learn more about the (TMP) and TMP to head width (HDW) or head length (HDL) evolutionary relationships amongst the organisms in this island Figure 1 – (a) Phylogenetic tree of 16s mitochondrial gene. Includes 30 Leptopelis species with geographic distribution by color and maximum female size. Closed circles at nodes is useful in our samples as well. We continue to explore new represent (>95% CI). Female size and distribution from (Schiotz et al 1999). (b) Biogeographical map of African ecosystems partitioned into 8 areas; Partitioned areas from (Linder et al. system. 2012). features that may be incorporated into the literature for future aid in identification. Variation in Leptopelis Species Complex Results Results In figure 1 (a), Leptopelis palmatus is clearly separated from the L. macrotis/ L. millsoni/ L. rufus complex with strong Leptopelis palmatus Conclusion support (>95% CI). There is not strong support for anything beyond the divergence of these groups, although it appears that there is a closer relationship with species of the Guinean-Congolian distribution. West and Central African Leptopelis species (L. rufus, L. macrotis, In figure 2 (a), we see clear phylogeographic structure across the species complex with support for distinct lineages of L. and L. millsoni) were previously thought to be the closest ancestors of macrotis in West Africa and L. millsoni in Central Africa, distinct lineages of L. millsoni on Bioko island, and pronounced L. palmatus due to both their large size and proximity to the island phylogeographic structure in L. rufus across Cameroon. chain. According to our most recent data, along with more extensive Leptopelis palmatus Photo credit: A. Stanbridge Photo credit: A. Stanbridge sampling from our collaborators, we find that L. palmatus is not closely related to any of the species mentioned above. Our results Phylogeographic Tree of West-Central African Sampling Map of West-Central African Leptopelis indicate that L. palmatus is within a group of Central African species Leptopelis Species Complex Species Complex that are smaller in body size, suggesting that there may be a b) convergence in gigantism within this genus. L_aubryi L_mills_Cam_ER_CAS253387 L_mills_Cam_ER_CAS255076 a) L_mills_Cam_ER_CAS255075 L_mills_Cam_ER_CAS255079 Moving forward, we are going to (1) increase our species and L_mills_Cam_ER_CAS255078! L_mills_Cam_SD_CAS253541 ! L_mills_Cam_ER_CAS255077 L_mills_Cam_ER_CAS255080 Cameroon - L_rufus_Cam_LR_CAS254044 L_rufus_Cam_LR_CAS254051East Region genetic sampling to determine which species of Leptopelis L. L_rufus_Cam_LR_CAS256680 ! L_rufus_Cam_LR_CAS256808 L_rufus_Cam_CR_CAS249982 L_rufus_Cam_CR_CAS249984 L_rufus_Cam_CR_CAS249981 ! L_rufus_Cam_CR_CAS249976 ! L_rufus_Cam_CR_CAS249983 palmatus is closely related to, and (2) collect ddRADseq data for the L_rufus_KX67 L_rufus_Cam_LR_CAS256879 L_rufus_Cam_LR_CAS256806 Leptopelis palmatus L_rufus_Cam_LR_CAS256807! ! Leptopelis rufus L_rufus_Cam_LR_CAS256877 L_rufus_Cam_LR_CAS256882 Photo credit: B. Freiermuth Photo credit: Rayna Bell L_rufus_Cam_SD_CAS254099! L. macrotis/ L. millsoni/ L. rufus complex and look at more L_rufus_Cam_SD_CAS254132 L_rufus_Cam_SD_CAS254131 L. rufus! L_rufus_Cam_LR_CAS256717 L_rufus_Cam_LR_CAS256683! L_rufus_Cam_LR_CAS254050 L_rufus_Cam_LR_CAS254048 L_rufus_Cam_LR_CAS254052 Cameroon - ! specimens to assess the morphological variation in these populations. L_rufus_Cam_LR_CAS256681 L_rufus_Cam_LR_CAS256682 L_rufus_Cam_LR_CAS256878 Littoral L_rufus_Cam_LR_CAS256718 L_rufus_Cam_LR_CAS254046 ! L_rufus_Cam_SD_CAS254100Region L_rufus_Cam_LR_CAS256880 These data will provide greater insight into the biogeography of land L_rufus_Cam_LR_CAS254045 L_rufus_Cam_SD_CAS254133 L_rufus_Cam_LR_CAS256823! ! L_rufus_Cam_LR_CAS253839 L_rufus_Cam_LR_CAS256881 L_rufus_Cam_LR_CAS256684 L_rufus_Cam_LR_CAS253968! bridge and oceanic islands in the Gulf of Guinea archipelago and L_rufus_Cam_LR_CAS254042 L_rufus_Cam_LR_CAS254047 L_rufus_Cam_BYU_62296 L_rufus_Cam_LR_CAS_256883 L_rufus_Cam_SWR_BYU_62290 L_rufus_Cam_SWR_BYU_62285! L_rufus_Gabon_MCZ_139662 ! diversification in the Guineo-Congolian forests. In addition, we hope L_rufus_Cam_LR_CAS_256876 L_rufus_Cam_LR_CAS_254049! L_rufus_Cam_SWR_BYU_62286 L_rufus_Cam_LR_CAS_253836 Methods L_rufus_Cam_LR_CAS_254043 Cameroon ! L_rufus_Cam_LR_BYU_62298 L_cf_rufus_Cam_MV_MH_0264 to find additional morphological characteristics to improve field L_cf_rufus_Cam_EF_MH_0242 L_rufus_Cam_SWR_BYU_62291 ! L_rufus_Nig_CR_MVZ_253290 ! L_rufus_Cam_Sd_CAS_254098 L_rufus_Cam_LR_CAS_253838! L_rufus_Cam_SWR_BYU_62293 L_rufus_Cam_LR_CAS_253837 identifications in this challenging species complex. L_cf_rufus_Cam_EF_MH_0215 L. millsoni! L_rufus_Cam_SWR_BYU_62288 L_rufus_Cam_SWR_BYU_62289 ! L_rufus_Bioko_NCSM_84364 ! L_rufus_Bioko_NCSM_84365 Bioko ! We collected 16s mitochondrial gene sequences for 33 species of L_rufus_Bioko_NCSM_84366 ! L_rufus_Gabon_MCZ_139814 ! L_rufus_Gabon_MCZ_139664 ! L_rufus_Gabon_MCZ_139665 ! L_rufus_Gabon_NCSM_76830 Gabon ! L_rufus_Gabon_MCZ_139713 ! BLS135601 ! Leptopelis and 146 individuals for our four target species. To L_cf_rufus_Cam_EF_MH_0421 ! L_cf_rufus_Cam_EF_MH_0422 L_rufus_Gabon_NCSM_77702 L_rufus_Gabon_NCSM_77701 L. Rufus L_macro_Ghan_WR_UWBM8890 L_macro_Ghan_A_ZMB_71318! L_mills_Cam_ER_CAS_253386 ! determine whether L. palmatus is closely related to the L. rufus/ L. L_mills_Cam_Sd_CAS_253581 Type Locality L_mills_Cam_Sd_CAS_253582 Ghana ! L_mills_Cam_Sd_CAS_253580 ! Acknowledgments L_millsoni_DCB L_rufus_Nig_CR_MVZ_253291 ! L_mills_Cam_SWR_MCZ_136851 ! Leptopelis millsoni L_rufus_Nig_CR_MVZ_253292 ! millsoni/ L. macrotis complex we generated a phylogeny using L_macro_Liber_GNF_GRE_40 L. Millsoni L_macro_Guin_H_PB_11_282 Photo credit: Rayna Bell L_macro_Lib_H_PLI_12_218 L_macro_Lib_H_PLI_12_373 ! L. macrotis! Type Locality We would like to thank our collaborators on the project: Adam Leaché, David Blackburn, Eli Greenbaum, Dan Portik, Greg L_macro_Guin_H_PB_11_383 L_macro_Lib_V_PLI_12_186 L_macro_Gui_FZ_CB2010_18! BEAST v 1.8.4 including one representative sequence for each L_macro_Guin_D_MH0038_16 Jongsma, Matt Fujita, and Mark Oliver-Rödel. We thank Gracia Gonzalez Porter and the L.A.B. for assistance with sequences and L_macro_Guin_Z_PB_11_742 ! L_macro_Lib_H_PLI_12_372 Liberia- L_macro_Guin_H_PB_11_085 L. Macrotis facilities. We thank the NHRE program coordinators at the NMNH, Smithsonian Institution for their contributions to the REU L_macro_Lib_H_PGL_13_045 L_macro_Guin_H_PB_11_642Guinea L_macro_Lib_H_PLI_12_380 ! species, with Astylosternus as an out-group. We assessed L_macro_Guin_H_PB_11_384 Type Localtiy program. This research was funded by NSF OCE: 1560088 and an ADS
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