LAURA BRUCE

HEDGES LAB MEETI NG

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