LAURA BRUCEHEDGES LAB MEETING

OCTOBER 4 , 2013

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, WestIndies + 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, tRNAMet, tRNAIle, tRNATrp, tRNAAla, tRNAAsn, tRNACys, tRNATyr, 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 electrophoresisDNA 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 compositelikelihood methods

Bootstrap values indicated (statisticalconfidence)

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. Haetianusmay 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

ColleaguesNational 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, 2383–2397.

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: 660–674.

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. 10:118–131.

Muñoz, 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