Table S1 Sample data for Hypseleotris sp. voucher specimens supplied by the Evolutionary Biology Unit, South Australian Museum. Microsatellite genotypes of these samples were used in PCA analysis (Fig. S2) for comparison with groupings obtained in the present study.

Table S2 Specimen data and Genbank accession numbers for mtDNA cytochrome b sequences following phylogenetic tree presented in Fig. 2.

Table S3 Microsatellite primers developed for genetic analysis of Hypseleotris spp.

Table S4 Example of allele profiles for a single site (HS) arranged by the four groupings defined by clustering analyses.

Fig. S1 Plot of Delta K calculated as the mean of the second-order rate of change in likelihood of K divided by the standard deviation of the likelihood of K, m(|L′′(K)|)/s[L(K)].

Fig. S2 Principal component analysis (PCA) performed on multilocus genotypes of 478 Hypseleotris spp. individuals from the present study and 27 voucher samples representing five different allozyme groups identified by Bertozzi et al. (2000).

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