EVOLUTION, GENETICS, & SYSTEMATICS From the Floor, to the Canopy: Comparative Phylogeography of Two Sympatric Chameleon Species in Central Africa’s Albertine Rift Poster 55 - Abstract 231 Daniel F. Hughes1-2, Wilber Lukwago3, Mathias Behangana3, Michele Menegon4, J. Maximilian Dehling5, Jan Stipala6, Colin R. Tilbury7, Krystal A. Tolley7-8, Chifundera Kusamba9, and Eli Greenbaum1-2 JMIH - 14 July 2017 Rio Grande Hall 1Department of Biological Sciences, 2UTEP Biodiversity Collections, University of Texas at El Paso, El Paso, Texas, USA; 3Makerere University, Kampala, Uganda; 4Museo Tridentino di Scienze Naturali, Trento, Italy; 5Universität Koblenz-Landau, Koblenz, Germany; 6University of Exeter, Cornwall, United Kingdom; 7University of Stellenbosch, Stellenbosch, South Africa; 8South African National Biodiversity Institute, Cape Town, South Africa; 9Centre de Recherche en Sciences Naturelles, Lwiro, République Démocratique du Congo Austin, TX Introduction Results Conclusions The Albertine Rift (AR) is one of the most diverse - Three new species of forest chameleons highland regions in continental Africa, yet it is not 1. Gene trees 2. Dated species trees 3. Phylogeography (Kinyongia) were formally described from the AR well known whether taxa with overlapping ranges (Hughes et al., 2017) and five new species of pygmy share diversification patterns. Kinyongia - PP > 0.95 + BS > 70% | - PP > 0.95 | - BS > 70% Kinyongia - PP > 0.95 | - 95% highest posterior density | mya - million years ago Kinyongia Rhampholeon chameleons (Rhampholeon) await description from the AR (Hughes et al., in review). Kinyongia adolfifriderici and Rhampholeon - K. adolfifriderici - K. adolfifriderici K. tolleyae < 1000 m AFRICA boulengeri are generally sympatric and share K. rugegensis sp. nov. º - The Miocene was identified as an important 5.1 mya 1000 - 1500 m 31 E habitats in the AR, yet they differ in microhabitat Area enlarged 1500 - 2000 m period for diversification of chameleons in the AR, use and foraging behavior. Rhampholeon / K. tolleyae 2000 - 3400 m R. sp. 1 which is linked to the fragmentation of African boulengeri is largely terrestrial and forages on the Lendu Plateau forests over this period (Kissling et al., 2012). 7.9 mya K. rugegensis 50 km N Lake forest floor, whereas K. adolfifriderici is mostly Albert - Diversification patterns for Kinyongia were arboreal and forages high in the forest canopy. *BEAST/BEAST / 1º N DEMOCRATIC consistent with the model of allopatric speciation We investigated the relative roles of K. tolleyae sp. nov. REPUBLIC driven by forest fragmentation in the Miocene. environmental and taxon-specific factors in K. itombwensis OF THE R. sp. 2 K. gyrolepis Rwenzori Mts. influencing spatiotemporal patterns of genetic 11.5 mya CONGO - Diversification events at the genus level for diversity in two sympatric chameleon species. North Rhampholeon generally fit the vicariance-driven / Orientale UGANDA Kivu Lake speciation model, however, it does not fully explain K. gyrolepis Edward Kigezi Highlands the rapid radiation across the Miocene–Pliocene K. adolfifriderici K. rugegensis 1º S R. sp. 3 Methods K. adolfifriderici 6.4 mya boundary in the R. boulengeri complex with several species distributions that span traditional Samples and sequencing. Forty-six samples of R. K. itombwensis sp. nov. / biogeographic barriers in the AR. RWANDA boulengeri K. adolfifriderici K. itombwensis Lake and 14 samples of were 0.03 Kivu - Several species in the R. boulengeri complex are K. gyrolepis R. sp. 4 collected during surveys in the AR from 2008–2016, Figure 1. Maximum-likelihood phylogeny of Kinyongia adolfifriderici from the Albertine Rift, Central Africa. Figure 3. Species tree with mean diversification dates for the Kinyongia adolfifriderici species complex. sympatric but not syntopic because they occur in including topotypic material. Two mitochondrial 3º S largely non-overlapping elevational zones for which Rugege Highlands (16S, ND2) and one nuclear (RAG-1) gene were South adaptations to novel physical factors (e.g., - PP > 0.95 + BS > 70% | - PP > 0.95 | - BS > 70% - PP > 0.95 | - 95% highest posterior density | mya - million years ago sequenced using standard protocols. Rhampholeon Rhampholeon Kivu temperatures or climatic niches) likely initiated DNA data. Chromatographs were interpreted in - R. boulengeri - R. boulengeri K. adolfifriderici R. sp. 5 processes leading to parapatric speciation. Itombwe Plateau BURUNDI SEQMAN PRO and alignments were made using R. sp. 1 - We found that the Gradient Speciation Hypothesis MUSCLE in MESQUITE. We made minor manual Maniema (i.e., parapatric speciation via niche differentiation) adjustments in MACCLADE. We used PHASE in TANZANIA Lake º is more appropriate to explain diversification in the DNASP to phase haplotypes for RAG-1. The phased R. sp. 1 5.3 mya 27º E Tanganyika 31 E R. boulengeri R. boulengeri complex. sequences were used in species-tree analyses. *BEAST/BEAST 0.7/ Figure 5. Map of the Albertine Rift showing sampled populations for Kinyongia (circles) and Rhampholeon (squares). - Traditional diversification scenarios are Gene trees. Maximum-likelihood analyses were inadequate to explain the high levels of cryptic conducted with the GTRGAMMA model in RAXML R. sp. 2 diversity we found in R. boulengeri and K. using the rapid bootstrap algorithm with 1,000 adolfifriderici, which differ in ecomorphology. replicates. Bayesian-inference analyses were 6.3 mya 4. Elevational zonation conducted in MRBAYES and run for 20 million R. sp. 2 - Comparative approaches to phylogeography are generations sampling every 1,000 with 25% 0.9/ Rhampholeon needed to assess both environmental and burn-in. We used PARTITIONFINDER to establish ecological factors promoting species diversity. R. sp. 3 R. sp. 1 the best model of evolution. The concatenated data 2500 m set included nearly all species per genus and analyses were run on the CIPRES Science Gateway. 5.7 mya R. sp. 2 2250 m References Species trees and divergence dating. We used R. sp. 3 R. sp. 4 7.2 mya 0.6/0.8 2000 m *BEAST in the program BEAST to estimate species Hughes, DF, et al. 2017. Integrative taxonomy of the Central R. sp. 3 trees for focal taxa. We used BEAST to estimate / 5.5 mya African forest chameleon, Kinyongia adolfifriderici (Sauria: 1750 m divergence dates with an uncorrelated log-normal Chamaeleonidae), reveals underestimated species diversity in the Albertine Rift. Zoological Journal of the Linnean Society relaxed clock model and a Yule prior. We included / R. boulengeri 0.4 0.3 R. sp. 4 (published online - 20 May 2017). all chameleon genera and 22 squamate taxa plus 1500 m . Rhampholeon boulengeri Sphenodon. Fossil calibrations were placed on nine R. sp. 4 R. sp. 5 Hughes, DF, et al Cryptic diversity in nodes and secondary calibrations on five nodes. We R. sp. 5 1250 m (Sauria: Chamaeleonidae), a pygmy chameleon from the Albertine Rift biodiversity hotspot. Molecular Phylogenetics analyzed five concurrent runs of 200 million R. sp. 5 1000 m and Evolution(in review - submitted 3 June 2017). generations sampled every 20,000. All runs were R. boulengeri combined using LOGCOMBINER with 10% burn-in. R. boulengeri Kissling, WD, et al. 2012. Cenozoic imprints on the 0.02 750 m phylogenetic structure of palm species assemblages *See Hughes et al. (2017) for detailed methods* worldwide. PNAS 109: 7379–7384. Figure 2. Maximum-likelihood phylogeny of Rhampholeon boulengeri from the Albertine Rift, Central Africa. Figure 4. Species tree with mean diversification dates for the Rhampholeon boulengeri species complex. Figure 6. Species tree with elevational zonation of six Rhampholeon species from the Albertine Rift, Central Africa..