Supporting Information Supplementary Methods Host genetics To verify that the contact zone between host subspecies is still located along the Rio Grande Valley near the San Acacia constriction between the towns of La Joya and San Acacia, New Mexico, 15 pocket gophers were examined from five locations, two north and three south of the San Acacia Constriction (Fig. 1; Table S1). Eight of the specimens were collected in 2011 and the other 7 were collected from 1990-1992. The New Mexico Department of Game and Fish (NMDGF) approved collection of specimens, and procedures followed all guidelines set by the University of Northern Iowa Institutional Animal Care and Use Committee and the American Society of Mammalogists (Sikes and Gannon 2011). DNA was extracted from pocket gopher tissues using the DNeasy Tissue Kit (Qiagen, Valencia, California) according to manufacturer specifications. The mitochondrial COI gene was amplified in a polymerase chain reaction (PCR) using primers CO1-5285F and CO1-6929R (Spradling et al. 2004) with Promega Hotstart Mastermix Mix (Promega, Madison, WI) according to manufacturer specifications with 40 thermal-cycles at an annealing temperature of 45C. PCR products were treated with ExoSAP-IT (USB, Cleveland, OH) and were sent to Iowa State University DNA Facility for sequencing with primer CO1-5285F on an Applied Biosystems 3730xl DNA Analyzer. Three nuclear loci were amplified and sequenced similarly. A portion of the first exon of the Interphotoreceptor Retinoid Binding Protein (IRBP) gene was amplified primers 119A (Jansa and Voss 2000) and 1297D (Jansa and Voss 2000, Stanhope et al. 1992) as described in Hafner et al. 2014. Both primers were used for sequencing. Amplification of a portion of the Recombination Activating Gene 1 (Rag1) was performed using primers Rag1-S70 and Rag1- S73 (Steppan et al. 2004) at an annealing temperature of 54C. A portion of the βfibrinogen gene was amplified using primers FIB-B17L (Prychitko and Moore, 1997) and R2 (Spradling et al. 2004) with an annealing temperature of 50C. Both primers were used for sequencing for all nuclear genes to better assess potential heterozygous sites. Sequences were screened for error and edited manually for heterozygosity using Geneious Pro (version 5.4.6, Biomatters Ltd), Heterozygous sequences in the three nuclear genes were duplicated and treated as two different alleles, each with one of the two alternative nucleotides. Because cloning to determine allele sequences for heterozygotes was beyond the needs of this analysis, where an individual was heterozygous at multiple nucleotide positions for a nuclear gene, existing homozygous genotypes of other individuals were used to guide nucleotide composition for each of the reconstructed alleles. Appropriate substitution models and uncorrected sequence divergence values were assessed for the four genes using Mega 6.06 and Bayesian Information Criterion scores (Tamura et al. 2013). Bayesian analysis was performed with the selected model using the MrBayes (Ronquist et al. 2012) plugin for Geneious R8 (http://www.geneious.com, Kearse et al. 2012). Chain length was set to 1,500,000 with 4 heated chains, a 0.1 heated-chain temperature, and a sub-sampling frequency of 300. Burn-in was set to 100,000 using unconstrained branch lengths for priors. Output was evaluated to assess the quality of runs using three criteria recommended in the MrBayes documentation (http://mrbayes.sourceforge.net/manual.php). Neighbor-joining trees also were built using similar molecular substitution models for comparison as the neighbor-joining trees depicted zero-branch lengths in a more intuitive manner. Louse microsatellite primer development A DNeasy Blood & Tissue Kit (Qiagen, Valencia, California) was used to extract DNA. From the pool of 160 lice. Manufacturer’s recommendations were followed with the following exceptions: lice were used directly from the ultra-cold freezer and use of liquid nitrogen was eliminated, after 4 hours of incubation in ATL buffer, an additional 20 µl Proteinase K was added with additional crushing performed before continued incubation overnight at 56ºC, final elution in AE Buffer was decreased from 200 µl to 50 µl, and incubation time was increased to 5 minutes. At Cornell University, pooled louse DNA was treated with the restriction enzyme Hinc III and then ligated to a double stranded SNX linker. The ligation procedure was modified to generate Pme I sites if linkers ligated to themselves. Digested, ligated fragments were enriched for microsatellites by hybridization to 3' biotinylated di-, tri-, and tetra-nucleotide repeat probes. PCR amplified products were ligated to 1.0 µl of a Titanium Rapid Library MID adapter (10 µm adapter stock), and small fragments were removed with Ampure beads. Libraries were submitted to the Sequencing and Genotyping Facility at Cornell Life Sciences Core Laboratory Center for FAM-quantification and Titanium 454 sequencing. Single sequence reads and contigs from analysis of 454 sequencing data were analyzed at University of Northern Iowa, and loci were chosen for amplification if they had a tetrameric repeat structure with a minimum of five repeats. Loci chosen based on multiple sequence reads were given "names"; loci chosen based on single sequence reads were numbered based on sequencing read number. Primerselect software (Lasergene Core Suite package, DNAStar, Madison, WI) was used to identify suitable primers to amplify each locus. A long M13 tag (5’CGAGTTTTCCCAGTCACGAC-3’) was added to the 5’ end of all locus-specific forward primers to allow concurrent amplification with a fluorescent primer (Schuelke 2000). A short tag (5'GTTTCTT-3') was added to all locus-specific reverse primers to promote adenylation and reduce stutter (Brownstein et al. 1996). Fluorescent tags (6-FAM, HEX, NED) were added to the 5’ end of universal M13 primers (5’-CGAGTTTTCCCAGTCACGAC-3’) to allow three-primer amplification of PCR products and subsequent multiplex genotyping (Schuelke 2000). Individual amplification of 7 target microsatellite loci in 275 chewing louse individuals was performed as described in the manuscript “Host behavior drives parasite genetics at multiple geographic scales: Population genetics of the chewing louse, Thomomydoecus minor”. In order to check the accuracy of genotyping assignments of individual lice and to estimate locus-specific genotyping error rates, 15 randomly chosen DNA samples out of an initial 157 samples (10%) were chosen for re-screening. At least two lice per host individual were included in this re-assessment in which new dye-labeled, 3-primer PCR was performed and amplified products were submitted to Iowa State University for analysis alongside a subset of previous PCR products. Locus quality was further evaluated by examining null allele frequencies for each of the 15 infrapopulations examined as estimated using the EM algorithm of Dempster et al. (1977) via the FreeNA program (Chapuis and Estoup 2007). Tests for genotypic disequilibrium between each pair of loci in each population were performed using Genepop 4.2 web version (Rousset 2008, http://genepop.curtin.edu.au/) using the likelihood-ratio test statistic to assess significance. Observed heterozygosity (HO) and expected heterozygosity (HE) were calculated in Arlequin (version 3.5.1.2, Excoffier and Lischer 2010) for the source population of microsatellite sequences using three infrapopulations combined from Las Nutrias, NM. Supplementary Results Host genetics Mitochondrial COI sequences from pocket gophers collected on opposite sides of the San Acacia constriction, where host subspecies previously came into contact (Smith et al. 1983; Table S1), sorted neatly in two different haplotype groups that displayed an average 8.0 % uncorrected sequence divergence between them (1,035 bp sequence data). One haplotype group was found exclusively in gophers collected north of the San Acacia constriction and the other was found exclusively in gophers collected south of it (Fig. S1). The 499 bp portion of -fib sequenced (461 bp of intron 7 and 38 bp of exon 8) had 8 variable nucleotide sites, 6 of which appeared heterozygous in one or more individuals. Bayesian and neighbor-joining analysis of putative allele sequences created two clusters. Pocket gopher alleles from individuals collected north of the San Acacia constriction formed one genetic cluster, and individuals from south of the contact zone formed the other, with two exceptions (Fig. S2). One allele from a southern gopher individual grouped with otherwise northern alleles and one allele from one northern individual grouped with otherwise southern alleles. The 874 bp region of IRBP sequenced included 15 nucleotide positions that were variable; eight of these positions were variable only due to autapomorphies. Of the seven potentially phylogenetically informative positions, three were diagnostic for all pocket gophers collected south of the San Acacia constriction. The other four positions were heterozygous in many of the pocket gophers collected north of the San Acacia constriction. Bayesian and neighbor-joining analysis of IRBP putative alleles yielded several genetic clusters, one consisting solely of alleles from individuals collected south of the San Acacia constriction and a larger and more genetically diverse cluster consisting of alleles from all northern individuals plus a single allele from a southern individual (Fig. S3). The 735 bp region of Rag1 sequenced included seven variable nucleotide positions, six of which were potentially phylogenetically informative. All pocket gophers collected north of the constriction had identical nucleotide sequences except for one unique nucleotide position observed in two individuals, and all northern individuals differed from all southern individuals by anywhere from one to four variable nucleotide positions. Bayesian and neighbor-joining analysis of Rag1 alleles indicated two genetic clusters that were consistent with geographic distribution, with the exception of a single southern allele that showed greatest resemblance to otherwise Northern alleles (Fig. S4). Taken together, mtDNA and all 3 nuclear genes show clear distinctions between haplotypes or alleles carried by pocket gophers from north of the constriction vs. south of it, yet there also were some exceptions to this pattern. Typically northern nuclear alleles were found in the southern localities of Lemitar (2 of 6 nuclear alleles, each from different individuals, sampled in 2011) and San Acacia (1 of 6 nuclear alleles sampled in 1990-1991), and typically southern alleles were found in the northern locality of Las Nutrias (1 of 9 nuclear alleles sampled in 2011). Quality of louse microsatellite loci For these 7 loci, accuracy testing showed no genotyping errors in which allele size was incorrectly identified in individuals after rescreening. Genotyping errors did occur for some of these loci when an individual louse originally amplified as a heterozygote and later amplified as a homozygote, or vice versa, but this non-repeatability was limited, yielding an allelic error rate as high as 7% for a single locus, but near 0% for other loci (Table S1). Among the 7 loci chosen for downstream analyses, there were no pairs of loci that showed evidence of genotypic disequilibrium across all populations after correction for multiple tests, and there were no loci that showed evidence of null alleles in any of the infrapopulations (Table S1). Observed and expected heterozygosity values were not significantly different for these loci at the source population from which the primer information was derived, and null alleles were estimated to be rare (0-.01-0.03 averaged over all 15 infrapopulations; Table S1). Supplementary Literature Cited Brownstein MJ, Carpten JD, Smith JR (1996) Modulation of non-templated nucleotide addition by Taq DNA polymerase: Primer modifications that facilitate genotyping. Biotechniques, 20, 1004-1006, 1008-1010. Chapuis MP, and Estoup A (2007) Microsatellite null alleles and estimation of population differentiation. Molecular Biology and Evolution, 24, 621-631. Goudet J (1995) FSTAT (version 1.2): a computer program to calculate F-statistics. Journal of Heredity, 86, 485-486. Goudet J (2001) FSTAT, a program to estimate and test gene diversities and fixation indices (version 2.9.3.2). Accessed online 23 March 2015 at: http://www2.unil.ch/popgen/softwares/fstat.htm. Hafner MS, Hafner DJ, Gonzáles EE, Demastes JW, Spradling TA, Cervantes FA (2014) Rediscovery of the pocket gopher Orthogeomys lanius (Rodentia: Geomyidae) in Veracruz, Mexico. Journal of Mammalogy, 95, 792-802. Jansa S, Voss RS (2000) Phylogenetic studies on didelphid marsupials I. Introduction and preliminary results from nuclear IRBP gene sequences. Journal of Mammalian Evolution, 7, 43-77. Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, Buxton S, Cooper A, Markowitz S, Duran C, Thierer T, Ashton B, Meintjes P, Drummond, A (2012) Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics, 28, 1647-1649. Prychitko TM, Moore WS (1997) The utility of DNA sequences of an intron from the -fibrinogen gene in phylogenetic analysis of woodpeckers (Aves: Picidae). Molecular Phylogenetics and Evolution, 8, 193-204. Rannala B (2012) BayesAss edition 3.0 user’s manual. Accessed online 23 March 2015 at: http://www.rannala.org/?page_id=245. Ronquist F, Teslenko M, van der Mark P, Ayres DL, Darling A, Höhna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP (2012) MrBayes 3.2: Efficient bayesian phylogenetic inference and model choice across a large model space. Systematic Biology, 61, 539-542. Schuelke M (2000) An economic method for the fluorescent labeling of PCR fragments. Nature Biotechnology, 18, 233-234. Sikes RS, Gannon WL (2011) Guidelines of the American Society of Mammaologists for the use of wild mammals in research. Journal of Mammalogy, 92, 235-53. Smith MF, Patton JL, Hafner JC, Hafner DJ (1983) Thomomys bottae pocket gophers of the central Rio Grande Valley, New Mexico: Local differentiation, gene flow, and historical biogeography. Occasional Papers The Museum of Southwestern Biology, 2, 1-15. Spradling TA, Brant SV, Hafner MS, Dickerson CJ (2004) DNA data support a rapid radiation of pocket gopher genera (Rodentia: Geomyidae). Journal of Mammalian Evolution, 11, 105125. Stanhope MJ, Czelusniak J, Si J-S, Nickerson J, Goodman M (1992) A molecular perspective on mammalian evolution from the gene encoding interphotoreceptor retinoid binding protein, with convincing evidence for bat monophyly. Molecular Phylogenetics and Evolution, 1, 148-160. Steppan SJ, Storz BL, Hoffmann RS (2004) Nuclear DNA phylogeny of the squirrels (Mammalia: Rodentia) and the evolution of arboreality from c-myc and Rag1. Molecular Phylogenetics and Evolution, 30, 703–719. Tamura K, Stecher G, Peterson, D, Flipski A, Kumar S (2013) MEGA6: Molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution, 30, 2725-2729. Wilson GA, Rannala B (2003) Bayesian inference of recent migration rates using multilocus genotypes. Genetics, 163, 1177-1191. Fig. S1. Pocket gopher host genetics based on Bayesian (HKY85 model) and neighborjoining analysis (Tamura-Nei model) of mitochondrial COI sequences yielded the same tree structure. Neighbor-joining analysis is shown here for its more intuitive representation of zero branch lengths. Gophers collected north of the contact zone are shown in blue and names are designated with “N”; gophers collected south of the zone are shown in red and names are designated with “S”. Individuals are identified with nearest town and collector number. Fig. S2. Pocket gopher host genetics based on Bayesian and neighbor-joining analysis (JC69 model) of nuclear -fibrinogen sequences with heterozygous sequences split into putative alleles based on other observed homozygous sequences. Both analyses yielded the same tree structure, but neighbor-joining analysis is shown here for its more intuitive representation of zero branch lengths. Gophers collected north of the contact zone are shown in blue and names are designated with “N”; gophers collected south of the zone are shown in red and names are designated with “S”. Individuals are identified with nearest town and collector number. Fig. S3. Pocket gopher host genetics based on Bayesian and neighbor-joining analysis (HKY85 model) of nuclear IRBP sequences with heterozygous sequences split into putative alleles based on other observed homozygous sequences. Both analyses yielded the same tree structure, but neighbor-joining analysis is shown here for its more intuitive representation of zero branch lengths. Gophers collected north of the contact zone are shown in blue and names are designated with “N”; gophers collected south of the zone are shown in red and names are designated with “S”. Individuals are identified with nearest town and collector number. Fig. S4. Pocket gopher host genetics based on Bayesian and neighbor-joining analysis (JC69 model) of nuclear Rag1 sequences with heterozygous sequences split into putative alleles based on other observed homozygous sequences. Both analyses yielded the same tree structure, but neighbor-joining analysis is shown here for its more intuitive representation of zero branch lengths. Gophers collected north of the contact zone are shown in blue and names are designated with “N”; gophers collected south of the zone are shown in red and names are designated with “S”. Individuals are identified with nearest town and collector number. Fig. S5. Chewing louse genetics based on Bayesian analysis (HKY85 model) of mitochondrial COI sequences from 275 T. minor and one outgroup. Host number is given first followed by louse individual number. Shortest branch lengths (e.g. between louse 436.09 and louse 436.23 from the same host) represent identical sequences. Lice collected on northern gophers are shown in blue (top clade); Lice collected on southern gophers are shown in red (bottom clade). For clarity, only representative specimen numbers are listed. Fig. S6. Chewing louse infrapopulation relationships based on a midpoint-rooted neighbor-joining tree built from DA values generated based on 7 microsatellite loci for 15 infrapopulations occurring on both sides of the San Acacia constriction (northern infrapopulations are indicated by blue branches and southern infrapopulations by red branches). Infrapopulations two major clusters consistent with geography (and host subspecies). Clustering pattern within each group analyzed in isolation (Figs. S6 and S7) differs slightly from the larger analysis shown here. Fig. S7. Chewing louse infrapopulation relationships based on a midpoint-rooted neighbor-joining tree built from DA values generated based on 7 microsatellite loci for 10 infrapopulations occurring north of the San Acacia constriction. Infrapopulations do not show clustering strictly consistent with locality or time. Fig. S8. Chewing louse infrapopulation relationships based on a midpoint-rooted neighbor-joining tree built from DA values generated based on 7 microsatellite loci for 5 infrapopulations occurring on south of the San Acacia constriction. Infrapopulations show clustering consistent with locality and, more weakly, with time. Table S1. Microsatellite loci, primer sequences, genetic diversity measures, and measures of locus reliability. Locus name 3495 Primer sequences (5’ 3’); primer tags are underlined, species-specific sequences are not underlined CGAGTTTTCCCAGTCACGACAATGAGTAAGTACGATCCAGCAC GTTTCTTAGTGAGTTAAATGCTAGGCTGATG 1569 CGAGTTTTCCCAGTCACGACTTTAACAAAGAGGAATCGGATGC GTTTCTTCATACTCCCTCACGATTCTGTCC 1451 CGAGTTTTCCCAGTCACGACCTCATGGTGATGGTCTTTGTCTC GTTTCTTGTGTTCGAGAAGCTGTATCATCC Jan CGAGTTTTCCCAGTCACGACAATTGTGCTATGTCATCACTTGG GTTTCTTCTTCAAACTTGCATTTACCTCTTC Allie CGAGTTTTCCCAGTCACGACACCACATACACTATCAGCAATCC GTTTCTTTTAATTCAAGGACCAGTGGAGGG Belle CGAGTTTTCCCAGTCACGACATCCCTCTTTCTGTCGTTGGAAG GTTTCTTTGCGGTTCGAAATTCTCCAAG 851 CGAGTTTTCCCAGTCACGACTTAATTCACACTCAATCCAGGCG GTTTCTTGTGTAGGCGTCCCACAAATTTG ano comparisons of Ho and He were significantly different (p = 0.17-1.0) brepresents 2 miscalled alleles in 15 diploid genotypes crepresents 1 miscalled allele in 15 diploid genotypes # alleles, and size of smallest allele (base pairs) HO and HE at the Las Nutrias, NM, localitya Estimated null allele frequency averaged over 15 infrapopulations 3 190 bp 2 198 bp 3 337 bp 2 198 bp 4 211 bp 4 215 bp 3 436 bp 0.53—0.59 0.02 Estimated allelic error rate based on sample reamplification and rescoring 0.00 0.16—0.20 0.03 0.07b 0.41—0.45 0.01 0.00 0.53—0.63 0.03 0.03c 0.28—0.37 0.02 0.00 0.25—0.26 0.02 0.00 0.41—0.40 0.03 0.03c Table S2. FST values calculated in FSTAT (version 2.9.3.2; Goudet 1995, 2001). FST (from FSTAT); highlighted values are significant at p < 0.009 (BY correction value) based on results of 2100 permutations LN751 LN752 LN753 LJ759 LJ760 LJ761 LJ434 LJ435 LJ436 LJ437 SA1428 SA3243 LE756 LE757 LN752 0.1055 LN753 0.0708 0.0857 LJ759 0.0503 0.0924 0.0920 LJ760 0.0934 0.0963 0.1175 0.0624 LJ761 0.0906 0.1287 0.0869 0.0302 0.0031 LJ434 0.0365 0.0381 0.0568 0.0110-0.0068 0.0078 LJ435 0.1203 0.1303 0.0532 0.0952 0.0408 0.0255 0.0534 LJ436 0.0663 0.0232 0.0742 0.0822 0.0418 0.0800 0.0195 0.0891 LJ437 0.0977 0.1289 0.1451 0.0337 0.0420 0.0094 0.0329 0.0686 0.0990 SA1428 0.4045 0.3567 0.3561 0.4823 0.3391 0.4126 0.3507 0.3263 0.2998 0.4527 SA3243 0.3940 0.3516 0.3586 0.4726 0.3130 0.3947 0.3261 0.3229 0.2526 0.4535 0.0629 LE756 0.5483 0.5737 0.5198 0.6238 0.4933 0.5498 0.5029 0.4558 0.4391 0.5915 0.1165 0.1871 LE757 0.4636 0.4637 0.4024 0.5415 0.4082 0.4734 0.4224 0.3827 0.3654 0.5174 0.0945 0.1645 0.0033 SO3203 0.4644 0.4568 0.4202 0.5442 0.4105 0.4747 0.4242 0.3949 0.3596 0.5079 0.0567 0.1353 0.0627 0.0429 Locality abbreviations: LN = Las Nutrias, LJ = La Joya (759-761 = 2011; 434-437 = 1992), SA = San Acacia, LE = Lemitar, and SO = Socorro Red values = FST comparisons between 1992 and 2011 at the same locality Bold, underlined, italics values = FST comparisons between contemporaneous infrapopulations at the same locality Table S3. Migration rate estimates from BayesAss 3.0 (Wilson and Rannala 2003) for migration into southern infrapopulations from northern infrapopulations (below diagaonal) and into northern infrapopulations from southern infrapopulations (above diagonal). Shading indicates comparisons of northern and southern infrapopulations (blue and yellow shading), within locality (peach or lilac shading), between localities (orange and violet shading). Un-shaded values were not considered as these comparisons involve both a spatial and a temporal component. Las Nutrias LaJoya new LaJoya new LaJoya new old LaJoya old LaJoya old LaJoya old LaJoya San Acacia San Acacia Lemitar Lemitar Socorro m[0][0]: m[1][0]: m[2][0]: m[3][0]: m[4][0]: m[5][0]: m[6][0]: m[7][0]: m[8][0]: m[9][0]: m[10][0] m[11][0] m[12][0] 0.7256 0.0187 0.0167 0.0155 0.018 0.0161 0.0142 0.0143 0.083 0.0111 0.0107 0.011 0.01 m[0][1]: m[1][1]: m[2][1]: m[3][1]: m[4][1]: m[5][1]: m[6][1]: m[7][1]: m[8][1]: m[9][1]: m[10][1]: m[11][1]: m[12][1]: 0.0583 0.6964 0.031 0.0572 0.0475 0.043 0.032 0.0443 0.0111 0.0111 0.0109 0.0105 0.0096 m[0][2]: m[1][2]: m[2][2]: m[3][2]: m[4][2]: m[5][2]: m[6][2]: m[7][2]: m[8][2]: m[9][2]: m[10][2]: m[11][2]: m[12][2]: 0.022 0.0416 0.6941 0.0439 0.0466 0.0291 0.034 0.0473 0.0127 0.0122 0.0115 0.0113 0.0118 in south from north 0.01322 in north from south 0.01325 within locality north 0.03484 within locality south 0.06498 between locality north 0.02522 between locality south 0.03263 over time at La Joya 0.03731 m[0][3]: m[1][3]: m[2][3]: m[3][3]: m[4][3]: m[5][3]: m[6][3]: m[7][3]: m[8][3]: m[9][3]: m[10][3]: m[11][3]: m[12][3]: 0.0201 0.0318 0.0553 0.6924 0.0381 0.0495 0.0303 0.0525 0.0119 0.0118 0.012 0.0108 0.0121 m[0][4]: m[1][4]: m[2][4]: m[3][4]: m[4][4]: m[5][4]: m[6][4]: m[7][4]: m[8][4]: m[9][4]: m[10][4]: m[11][4]: m[12][4]: 0.0328 0.0741 0.0382 0.0454 0.6959 0.0318 0.0693 0.0354 0.013 0.0123 0.0112 0.0112 0.0113 m[0][5]: m[1][5]: m[2][5]: m[3][5]: m[4][5]: m[5][5]: m[6][5]: m[7][5]: m[8][5]: m[9][5]: m[10][5]: m[11][5]: m[12][5]: 0.0131 0.0182 0.0254 0.0209 0.0188 0.6941 0.0134 0.0181 0.0111 0.0115 0.0113 0.0115 0.0099 m[0][6]: m[1][6]: m[2][6]: m[3][6]: m[4][6]: m[5][6]: m[6][6]: m[7][6]: m[8][6]: m[9][6]: m[10][6]: m[11][6]: m[12][6]: 0.0232 0.0244 0.0314 0.021 0.0247 0.0265 0.7127 0.0229 0.0126 0.0133 0.0133 0.0125 0.0125 m[0][7]: m[1][7]: m[2][7]: m[3][7]: m[4][7]: m[5][7]: m[6][7]: m[7][7]: m[8][7]: m[9][7]: m[10][7]: m[11][7]: m[12][7]: 0.0405 0.0388 0.0253 0.0382 0.0297 0.0471 0.0286 0.7123 0.0106 0.0108 0.0118 0.0104 0.0095 m[0][8]: m[1][8]: m[2][8]: m[3][8]: m[4][8]: m[5][8]: m[6][8]: m[7][8]: m[8][8]: m[9][8]: m[10][8]: m[11][8]: m[12][8]: 0.0144 0.0119 0.016 0.015 0.0165 0.0153 0.0125 0.0111 0.6957 0.041 0.0224 0.0161 0.0196 m[0][9]: m[1][9]: m[2][9]: m[3][9]: m[4][9]: m[5][9]: m[6][9]: m[7][9]: m[8][9]: m[9][9]: m[10][9]: m[11][9]: m[12][9]: 0.0141 0.0123 0.0204 0.0173 0.0156 0.0142 0.0197 0.0103 0.0636 0.7604 0.0269 0.0213 0.0362 m[0][10]: m[1][10]: m[2][10]: m[3][10]: m[4][10]: m[5][10]: m[6][10]: m[7][10]: m[8][10]: m[9][10]: m[10][10]: m[11][10]: m[12][10]: 0.0106 0.0105 0.0168 0.0108 0.0187 0.0108 0.0108 0.0097 0.0698 0.0434 0.784 0.104 0.1132 m[0][11]: m[1][11]: m[2][11]: m[3][11]: m[4][11]: m[5][11]: m[6][11]: m[7][11]: m[8][11]: m[9][11]: m[10][11]: m[11][11]: m[12][11]: 0.0113 0.0107 0.0163 0.0114 0.0167 0.0117 0.012 0.0102 0.0307 0.0324 0.0513 0.7446 0.0334 m[0][12]: m[1][12]: m[2][12]: m[3][12]: m[4][12]: m[5][12]: m[6][12]: m[7][12]: m[8][12]: m[9][12]: m[10][12]: m[11][12]: m[12][12]: 0.014 0.0105 0.0132 0.0112 0.0131 0.0108 0.0103 0.0114 0.0461 0.0286 0.0228 0.0248 0.711 Table S4. Microsatellite analyses for individual infrapopulations. For each infrapopulation, collecting locality relative to the San Acacia constriction (N = north, S = south), collector number, and approximate locality is given. A) Total number of alleles sampled, B) Allelic richness and average allelic richness, C) Observed (Ho) and expected (He) heterozygosity, and D) Inbreeding coefficient (FIS).