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Members of Clibanarius virescens show considerable intraspecific colour variation, including colouration of the second/third pereopods (green/white) and the dactyls of the second/third pereopods (with or without dark bands/patches). However, factors inducing these colour variations have not yet been elucidated. Here, we investigated the occurrence of colour variation in this species with particular emphasis on change of colouration associated with growth stage and region in specimens from tropical/subtropical to warm temperate areas in the Indo–West Pacific, including evidence from molecular phylogeny based on the cytochrome c oxidase subunit I (COI). We have, then, clarified that the colouration on the pereopod dactyls gradually changed from solid colour (yellow/white) to having dark-coloured area(s) or transverse band(s) as a result of the growth stage. The frequency of occurrence of the solid colour dactyls was higher than those of other colour types in tropical regions. Our results also indicated that the white ambulatory leg type was the colouration type that was frequently seen in juvenile stages. However, significant genetic differences were not detected between each colouration determined by molecular analysis of samples from 14 localities in the Indo-West Pacific region; in contrast, two genetically differentiated regional populations (North Australia; Phuket, Thailand; and Lombok, Indonesia) were detected. The present study, therefore, emphasizes the necessity for further study on the colour variation of marine animals focusing on growth stages and regional differences, with molecular data to facilitate the research on adaptation and/or speciation, especially in geographically widely distributed species.

Species of hermit crabs in the genus Clibanarius Dana, 1852 have adapted to various environments in the intertidal areas, including hard substrates and soft sediments. These species often bear a close morphological resemblance to each other, therefore, the colouration on the pereopods can be one of the reliable characteristics to distinguish the species. However, the evolutionary relationships among species with different colour patterns and relationships between colour patterns and habitat adaptation have not previously been investigated. Therefore, we reconstructed the phylogenetic relationships among 19 species of Clibanarius based on mitochondrial [12S rRNA, 16S rRNA and cytochrome oxidase I] and nuclear [histone H3] DNA markers. The results suggest that the striped and solid colour elements have evolved multiple times independently, with the ancestral colour pattern potentially being scattered, bright colour spots with a bright colour band. Our findings also suggest that evolutionary adaptation from hard substrates to mudflats and soft sediments may have occurred at least twice.