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Figure 1.

Map showing distribution range of RLTBs and rivers from where samples were collected.

Map showing distribution range of RLTBs and rivers from where samples were collected, the species-tree built in *BEAST is shown on the left side. Posterior probability values below 1 are shown at the nodes. Photograph of a specimen considered as Puntius denisonii is shown; notice the absence of a black spot on the dorsal fin which is the current diagnostic character for distinguishing it from its congener Puntius chalakkudiensis found at location CHD in the map, the tip label codes are explained in Table 1. Note that according to the current taxonomy of RLTBs P. denisonii (most probably population PER-D), is nested within the different populations of P. chalakkudiensis, the other populations of P.denisonii are distributed above the palaghat gap. Here we show that each of these populations (labeled) qualify as evolutionarily distinct lineages.

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Table 1.

Micro-level distribution of the eight evolutionarily distinct lineages (EDL) including the two recognized species of RLTBs in the Western Ghats.

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Figure 2.

MANOVA/CVA discriminating different RLTB populations.

MANOVA/CVA of 24 size adjusted biometric characters of 12 RLTB populations. (a) Clusters of all 12 populations on the first two canonical axis and (b) pair wise matrix of Fisher’s distances between the centroids of the clusters (upper diagonal) and P values for Fisher’s distances (lower diagonal). Percent discrimination by each canonical axis is shown in parenthesis.

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Figure 3.

Results of the DNA based species delimitation methods.

Results of the DNA based species delimitation methods a) Heat-map showing the fixed distance threshold method and the clustering of the specimens; b) results of the GMYC method implemented on the cytb and cox1+ cytb ultrametric trees, the coloured blocks on the right side indicates the tips clustered together by the program as a same (putative) species; c) Result of the Bayesian method applied on the RLTB species tree, values at nodes are the speciation probabilities using three different prior settings (see materials and methods for details); each evolutionarily distinct lineage (tip) is denoted with a distinct colour.

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Figure 4.

Results of Divergence time analysis.

Timetree showing the divergence times of the major RLTB lineages, node bars denote the 95% credibility interval; values at nodes indicate the mean age in million years.

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