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Research supported by the
Martin, B.T., Bernstein, N.P., Birkhead, R.D., Koukl,
J.F., Mussmann, S.M., Placyk Jr., J.S. 2013. Sequence-based
molecular phylogenetics and phylogeography of the American box turtles
(Terrapene spp.) with support from DNA barcoding.
Molecular Phylogenetics and Evolution 68(2013)119–134.
The classification of the American box turtles (Terrapene
spp.) has remained enigmatic to systematists. Previous
comprehensive phylogenetic studies focused primarily on
morphology. The goal of this study was to re-assess the
classification of Terrapene spp. by obtaining DNA
sequence data from abroad geo- graphic range and from all four
recognized species and 11subspecies within the genus.Tissue
samples were obtained for all taxa except for Terrapene
nelsoni klauberi . DNA was extracted,and the mitochondrial
DNA (mtDNA) cytochrome b (Cytb) and nuclear DNA (nucDNA)
glyceraldehyde-3-phosphate-dehydrogenase (GAPD) genes were
amplified via polymerase chain reaction and sequenced. In
addition,the mtDNA gene commonly used for DNA barcoding
(cytochrome oxidasec subunit I; COI) was amplified and sequenced
to calculate pairwise percent DNA sequence divergence comparisons
for each Terrapene taxon. The sequence data were analyzed
using maximum likelihood and Bayesian phylogenetic inference,a
molecular clock, AMOVAs, SAMOVAs, haplotype networks,and pairwise
percent sequence divergence comparisons. Terrapene carolina
mexicana and T. c.yucatana formed a monophyletic
clade with T. c.triunguis , and this clade was
paraphyletic to the rest of T. carolina . Terrapene
ornata ornata and T. o.luteola lacked distinction
phylogenetically, and Terrapene nelsoni was confirmed to
be the sister taxon of T. ornata . Terrapene c. major , T. c.
bauri , and Terrapene coahuila were not well
resolved for some of the analyses.The DNA barcoding results
indicated that all taxa were different species (>2%sequence
divergence)except for T. c.triunguis – T. c.mexicana and T.
o.ornata – T. o.luteola . The results suggest that T.
c.triunguis should be elevated to species status (Terrapene
mexicana), and mexicana and yucatana should
be included in this group as subspecies. In addition, T.
o.ornata and T. o.luteola should not be considered
separate subspecies.The DNA barcoding data support these
recommended taxonomic revisions. Because conservation efforts are
typically species-based, these results will be important for
facilitating successful conservation management strategies.
http://www.sciencedirect.com/science/article/pii/S1055790313001073
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