OCTOBER 4, 201 3 Phylogenetic Analysis of the Diversification in Hispaniolan Trunk-Ground Anoles: Anolis cybotes species group Anolis cybotes Greater Antilles region Background: Anolis cybotes Anolis genus SE North America, Central South America, West Indies + satellite islands Small lizards Notable morphology: large heads and dewlaps Green Anoles: first reptile to have genome sequenced
Anolis cybotes Widespread cybotoid species group trunk-ground ecomorph class: low perches
A. doris Background: Anolis cybotes Anolis cybotes Evidence of evolutionary radiation of the cybotoid ecomorph based on adaptation to geographically distinct macrohabitats (forests vs. outcrops, etc.) Macrohabitat refers to vegetation, climate, topography
Very closely related in ecology and morphology A. cybotes, A. armouri, A. whitemani, A. longitibialis, A. haetianus, A. marcanoi, A. strahmi, A. shrevei
Extent of such species divergence largely undocumented New species? Taxonomical revisions necessary?
Previous Work (Glor et al.) Glor et al. Phylogenetic analysis of ecological and morphological diversification in Hispaniolan trunk-ground anoles (Anolis cybotes group). (2003)
112 samples from Hispaniola, across 68 localities A. cybotes A. whitemani A. armouri A. longitibialis A. marcanoi A. strahmi A. shrevei A. haetianus Phylogenetic analysis (mtDNA for ND2, tRNA Met , tRNA Ile , tRNA Trp , tRNA Ala , tRNA Asn , tRNA Cys , tRNA Tyr , ND1, COI)
Morphological analysis (leg length, toe pads, scales, etc.) Previous Work Previous Work (continued) Revealed that macrohabitat type and morphology are strongly related, independent of phylogenetic relatedness Species-level divergence evident Macrohabitat elicits adaptive responses via directional selection
My Project Expands upon Glor et al. (2003) Introduces 62 new cybotoid specimens to the data set Previously collected on lab excursions Across ~25 localities in Hispaniola and satellite islands
A. cybotes A. cybotes Methods Total genomic DNA extraction from frozen tissues QIAGEN DNeasy kit Polymerase Chain Reaction (PCR) amplification of mitochondrial DNA ND2 gene: two parts (~1200 bp total) L4437 + H4980 primers (~550 bp) 1 L4882a + H5617a
primers (~650 bp) 1
Gel electrophoresis DNA purification
1 Macey et al., 1997, 1998 Methods (continued) Sequence data analysis Aligned, evaluated, edited in MEGA V5.1 Phylogenetic tree Analyzed via neighbor-joining and maximum composite likelihood methods Bootstrap values indicated (statistical confidence) Results Glor et al. (2003) sequence data imported from GenBank Compiled with new specimen data
Large phylogeny constructed Several clades are notable Results (continued) A. Haetianus may be synonym for the true A. cybotes New species: A. notialis sp. nov. (indicating occurrence on southern side of peninsula) Results (continued) Likely to be full species A. doris Results (continued) Likely to be full species A. breslini True A. whitemani Results (continued) Possible new species, identified as A. sp. nov. for now Conclusions
Evidence of speciation based on phylogenetic data Taxonomy revisions needed Important: Since A. haetianus clade is likely the true A. cybotes, all other A. cybotes will need new name (A. riisei?) Morphological analyses needed to evaluate further
A. cybotes Hispaniola Current/Future Endeavors Conduct further phylogenetic analysis Rhodopsin nuclear gene for comparison and confirmation of suspected results Primers ROD3FGlor 2 and ROD4R (newly designed)
Currently working towards successful amplification Try another fast evolving nuclear gene
Morphological analysis Identify the visible differences between the specimens
Add more specimens to data set
2 Glor et al., 2003, 2004 Acknowledgements S. Blair Hedges Provided samples, animal images, research guidance Colleagues National Science Foundation (NSF) Critical Ecosystem Partnership Fund (CEPF)
Questions? A. cybotes References
Glor, R. E., Kolbe, J. K., Powell, R., Larson, A. & Losos, J. B. 2003 Phylogenetic analysis of ecological and morphological diversification in Hispaniolan trunk-ground anoles (Anolis cybotes group).Evolution 57, 23832397. Glor, Richard E., et al. "Partial island submergence and speciation in an adaptive radiation: a multilocus analysis of the Cuban green anoles." Proceedings of the Royal Society of London. Series B: Biological Sciences 271.1554 (2004): 2257-2265. Glor, Richard E., Jonathan B. Losos, and Allan Larson. "Out of Cuba: overwater dispersal and speciation among lizards in the Anolis carolinensis subgroup." Molecular Ecology 14.8 (2005): 2419- 2432. APA Macey, J. R., A. Larson, N. B. Ananjeva, and T. J. Papenfuss. 1997. Evolutionary shifts in three major structural features of the mitochondrial genome among iguanian lizards. J. Mol. Evol. 44: 660674. Macey, J. R., J. A. Schulte, N. B. Ananjeva, A. Larson, N. RastegarPouyani, S. M. Shammakov, and T. J. Papenfuss. 1998. Phylogenetic relationships among agamid lizards of the Laudakia caucasia species group: testing hypotheses of biogeographic fragmentation and an area cladogram for the Iranian Plateau. Mol. Phylogenet. Evol. 10:118131. Muoz, Martha M. and Hopwood, Juanita M., editors. Anolis cybotes." Encyclopedia of Life, available from http://eol.org/pages/795854/. Accessed 01 Oct 2013.
*URLs for images in slide comments, all animal images provided by S. Blair Hedges