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FIGURE 12 in Integration of molecular, bioacoustical and morphological data reveals two new cryptic species of Pelodytes (Anura, Pelodytidae) from the Iberian Peninsula

FIGURE 12. Dorsal, ventral and dorsolateral view, and close up of ventral side of forelimb, of living Pelodytes hespericus sp. nov.: (a-b) male holotype JDR 2012-1, (c-f); male paratype JDR 2012-2. Not to scale.

FIGURE 5 in Integration of molecular, bioacoustical and morphological data reveals two new cryptic species of Pelodytes (Anura, Pelodytidae) from the Iberian Peninsula

FIGURE 5. Ventral views of specimen hands (palmar surface) of the four western Pelodytes lineages. The pictures show variation in the size and arrangement of the subarticular and metacarpal tubercles within species and populations, suggesting that no fully diagnostic interspecific differences in this character exist. All photos from specimens in life. M, male, F, female. Asterisks denote mirrored images. Not to scale.

FIGURE 3 in Integration of molecular, bioacoustical and morphological data reveals two new cryptic species of Pelodytes (Anura, Pelodytidae) from the Iberian Peninsula

FIGURE 3. Clustering results of Bayesian assignment analyses using Structure, based on nuclear DNA sequences with the number of clusters K ranging from 1 to 7. Each individual is represented as a vertical bar partitioned into K colored segments, whose length is proportional to the individual's estimated group membership coefficient. The enlarged figure shows the results for K=4, with population numbers as in Fig. 1 and Supplementary Table 1. Population numbers on the top are shown to highlight individuals with mixed ancestry, and the mitochondrial haplogroup (A-D) for each included individual is shown in the bottom. Individuals in the graph are ordered as in Supplementary Table S1. Note that the comparatively large number of putative hybrids is due to the overrepresentation of populations in the contact zones in our analysis, as compared with Díaz- Rodríguez et al. (2015).

FIGURE 11 in Integration of molecular, bioacoustical and morphological data reveals two new cryptic species of Pelodytes (Anura, Pelodytidae) from the Iberian Peninsula

FIGURE 11. (a,b) Dorsal and ventral views of preserved Pelodytes atlanticus sp. nov. holotype (male; specimen GLA-03). (c,d) Dorsolateral view of paratype (GLA-02) and holotype (GLA-03) in life. (e-i) dorsolateral, dorsal, and ventral view, as well as ventral view of hand and foot of male paratype ZSM 194/2016. Not to scale.

FIGURE 10 in Integration of molecular, bioacoustical and morphological data reveals two new cryptic species of Pelodytes (Anura, Pelodytidae) from the Iberian Peninsula

FIGURE 10. Comparative photos of Pelodytes ibericus in life, male above, female below, from near Jerez de la Frontera, Spain. Specimens not collected. Images not to scale.

FIGURE 8 in Integration of molecular, bioacoustical and morphological data reveals two new cryptic species of Pelodytes (Anura, Pelodytidae) from the Iberian Peninsula

FIGURE 8. Comparative photos of Pelodytes punctatus in life. The upper two photos show a male and a female from near Montpellier, France; the lower two pictures show a male and a female from near Vic, Girona, Spain. Specimens not collected. Images not to scale. Identity of P. punctatus individuals assessed by geographical location; specimens not DNA barcoded.

FIGURE 2 in Integration of molecular, bioacoustical and morphological data reveals two new cryptic species of Pelodytes (Anura, Pelodytidae) from the Iberian Peninsula

FIGURE 2. Median-joining networks of Western Palearctic Pelodytes based on six nuclear markers and mtDNA (mt-ND4+mt- CYB). Samples in the nDNA networks are colored according to their assignment to the four major clusters in the mtDNA network, corresponding to the species as color coded in Fig. 1. Each circle represents a different haplotype and its size is proportional to its relative frequency. Black dots represent inferred unsampled or extinct haplotypes. Markers RPL3 and Cherpint7 are represented at the same scale.

FIGURE 7 in Integration of molecular, bioacoustical and morphological data reveals two new cryptic species of Pelodytes (Anura, Pelodytidae) from the Iberian Peninsula

FIGURE 7. Bioacoustic differences between lineages of Pelodytes occurring in the Iberian Peninsula. For raw data, see Supplementary Table S3. The boxplot shows the average number of B notes per call series, i.e, every call consisting of one A note and one or more B notes. Average values were first computed per individual, and then summarized across species. Differences are significant between P. hespericus sp. nov. and all other species, and between P. atlanticus sp. nov. and P. ibericus (ANOVA with Tukey's post-hoc test; see Results).

Figure 9 in Species delimitation in the Andean grasshopper genus Orotettix Ronderos & Carbonell (Orthoptera: Melanoplinae): an integrative approach combining morphological, molecular and biogeographical data

Figure 9 Orotettix males. Phallic complex, species as indicated. A, D, G, J, M, distal portion of aedeagal valves, lateral view; B, E, H, K, N, distal portion of aedeagal valves, dorsal view; C, F, I, L, O, epiphallus, dorsal view. Numbers indicate characters and states used in the phylogenetic analyses.

Figure 11 in Species delimitation in the Andean grasshopper genus Orotettix Ronderos & Carbonell (Orthoptera: Melanoplinae): an integrative approach combining morphological, molecular and biogeographical data

Figure 11 Outgroup taxa used in the phylogenetic analyses, species as indicated. A–D, male habitus. Scale bars: 5 mm. Numbers indicate characters and states used in the phylogenetic analyses.

Figure 12 in Species delimitation in the Andean grasshopper genus Orotettix Ronderos & Carbonell (Orthoptera: Melanoplinae): an integrative approach combining morphological, molecular and biogeographical data

Figure 12 Outgroup taxa used in the phylogenetic analyses, species as indicated. A, C, E, G, male distal abdominal segments, dorsal view; B, D, F, H, male distal abdominal segments, lateral view. Numbers indicate characters and states used in the phylogenetic analyses.

Figure 7 in Species delimitation in the Andean grasshopper genus Orotettix Ronderos & Carbonell (Orthoptera: Melanoplinae): an integrative approach combining morphological, molecular and biogeographical data

Figure 7 Orotettix males, species as indicated. A, C, E, G, I, distal abdominal segments, lateral view; B, D, F, H, J, distal abdominal segments, dorsal view. Numbers indicate characters and states used in the phylogenetic analyses.

Figure 4 in Species delimitation in the Andean grasshopper genus Orotettix Ronderos & Carbonell (Orthoptera: Melanoplinae): an integrative approach combining morphological, molecular and biogeographical data

Figure 4 Combined molecular and morphological phylogenetic analysis under parsimony criteria, using an extended implied weighting strategy. Numbers indicate branch supports (symmetric resampling).

Figure 5 in Species delimitation in the Andean grasshopper genus Orotettix Ronderos & Carbonell (Orthoptera: Melanoplinae): an integrative approach combining morphological, molecular and biogeographical data

Figure 5 Orotettix, phallic complex, in lateral (A) and dorsal views (B). Abbreviations: Ap, apodemes of cingulum; Ar, arch of aedeagus; Av, aedeagal valves; Ep, endophallic plates; E, epiphallus; L, lophi of epiphallus; Rm, rami; Sh, sheath of aedeagus.

Figure 13 in Species delimitation in the Andean grasshopper genus Orotettix Ronderos & Carbonell (Orthoptera: Melanoplinae): an integrative approach combining morphological, molecular and biogeographical data

Figure 13 Outgroup taxa used in the phylogenetic analyses, species as indicated. Phallic complex. A, D, G, J, distal portion of aedeagal valves, lateral view; B, E, H, K, distal portion of aedeagal valves, dorsal view; C, F, I, L, epiphallus, dorsal view. Numbers indicate characters and states used in the phylogenetic analyses.

Figure 3 A in Species delimitation in the Andean grasshopper genus Orotettix Ronderos & Carbonell (Orthoptera: Melanoplinae): an integrative approach combining morphological, molecular and biogeographical data

Figure 3 A, most parsimonious tree of the genus Orotettix (length 55, CI = 0.87, RI = 0.97) resulting from the cladistic analysis of the morphological character dataset, under equal weights. Black circles indicate unique changes and white circles indicate homoplasies. The numbers below the nodes are bootstrap support values, and those above are Bremer support values. The new species (O. sp. 1; O. sp. 2; O. sp. 3; O. sp. 4; O. sp. 5) delimited in this study based on molecular, morphological and geographical analyses are indicated in the tree. Lateral bars indicate the distribution of the specimens according to the geomorphic units of the Andes delimited by Gonzalez & Pfiffner (2012). B, geomorphological units of the Andes delimited by Gonzalez & Pfiffner (2012).

Figure 8 in Species delimitation in the Andean grasshopper genus Orotettix Ronderos & Carbonell (Orthoptera: Melanoplinae): an integrative approach combining morphological, molecular and biogeographical data

Figure 8 Orotettix males, species as indicated. A, C, E, G, I, distal abdominal segments, lateral view; B, D, F, H, J, distal abdominal segments, dorsal view. Numbers indicate characters and states used in the phylogenetic analyses.

Figure 10 in Species delimitation in the Andean grasshopper genus Orotettix Ronderos & Carbonell (Orthoptera: Melanoplinae): an integrative approach combining morphological, molecular and biogeographical data

Figure 10 Orotettix males. Phallic complex, species as indicated. A, D, G, J, M, distal portion of aedeagal valves, lateral view; B, E, H, K, N, distal portion of aedeagal valves, dorsal view; C, F, I, L, O, epiphallus, dorsal view. Numbers indicate characters and states used in the phylogenetic analyses.

Figure 2 in Species delimitation in the Andean grasshopper genus Orotettix Ronderos & Carbonell (Orthoptera: Melanoplinae): an integrative approach combining morphological, molecular and biogeographical data

Figure 2 Bayesian phylogenetic analysis of COI characters. Acronyms of specimens according to Table 1. Numbers on branches indicate posterior probabilities. Numbers in parentheses indicate bootstrap supports of maximum-parsimony analysis. Results of General Mixed Yule-coalescent (GMYC) analysis are represented as lateral bars with different line patterns. Each bar indicates a different cluster identified by GMYC. Solid black patterns indicate clusters which coincide with species delimitation based on results from the other molecular, morphological and geographical analyses. 1, O. andeanus; 2, O. sp. 1; 3, O. sp. 2; 4, O. hortensis; 5, O. sp. 3; 6, O. sp. 4; 7, O. carrascoi; 8, O. sp. 5; 9, O. ceballosi.

Figure 6 in Species delimitation in the Andean grasshopper genus Orotettix Ronderos & Carbonell (Orthoptera: Melanoplinae): an integrative approach combining morphological, molecular and biogeographical data

Figure 6 Orotettix males, species as indicated. A–J, habitus. Scale bars: 5 mm. Numbers indicate characters and states used in the phylogenetic analyses.