A New Green Salamander in the Southern Appalachians: Evolutionary History of Aneides aeneus and Implications for Management and Conservation with the Description of a Cryptic Microendemic Species

Austin Patton, Joseph J. Apodaca, Jeffrey D. Corser, Christopher R. Wilson, Lori A. Williams, Alan D. Cameron, and David B. Wake

Copeia 107: 748–763

Figure S1: Averaged model fits (DIC) across values of K (assumed number of populations) using TESS. Averages are across the five independent runs. The best fit model is selected by identifying the value of K at which the improvement in model fit (decrease in DIC) ‘elbows.’ In our case, this value of K is unambiguously four.

Figure S2: Mitochondrial phylogeny inferred using BEAST2 with 1) confidence intervals on divergence time estimations, and 2) labeled tips.

Figure S3: Map of samples used in mitochondrial phylogenetic analyses. This is the same map as in Figure 2, but individual points are labeled so as to enable comparison to Figure S2.

Figure S4: Nuclear phylogenies as estimated by RAxML without correcting for ascertainment bias. For this analysis, we did not exclude heterozygous sites. Filled circles denote nodes with support values >95%, empty circles denote nodes with support values >90%, and all other nodal support values are indicated as text. Numbered/colored circles enclosing monophyletic groups in the trees indicate species as identified by PTP when applied to the RAxML tree. Support for these species are as follows: 1) 57%, 2) 97%, 3) 95%.

Figure S5: Boxplots illustrating measurements of 20 morphological characters for each A. aeneus and A. caryaensis. Each trait is shown in its own column, and boxplots are colored by species. Significance values as two-tailed t-tests assuming heteroscedasticity conducted for each trait among species are shown above boxplots. Comparisons were made using transformed variables. Refer to Table 1 for units.

Table S1: Information pertaining to the 13 microsatellite loci developed for the Green Salamander, Aneides aeneus (Castaneides). N represents the number of individuals for which genotypes were obtained, k represents the number of alleles observed for each locus, and Freq Null is the frequency of null alleles across all samples. HO represents the observed heterozygosity, and HE represents the expected heterozygosity. HWE is the probability that each locus is in Hardy-Weinberg Equilibrium across all samples.

Table S2: Frequency of null alleles as estimated by Genepop at the 12 microsatellite loci used in this study within each population identified by TESS.

Table S3: Morphological measurements for samples of Aneides aeneus and A. caryaensis measured in this study. Refer to Table 4 for units.

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