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A long-spined sea urchin    Diadema   -sp reported from Japanese waters was genetically distinct from all known    Diadema    species, but it remained undescribed. Extensive field surveys in Japan with molecular identification performed in the present study determined five phenotypes (I to V) in    Diadema   -sp according to the presence and/or shape of a white streak and blue iridophore lines in the naked space of the interambulacral area. All phenotypes were distinct from    Diadema   setosum    (Leske, 1778) and    Diadema   savignyi    (Audouin, 1829), of which a major type (I) corresponded to    Diadema   clarki    Ikeda, 1939 that was questioned and synonymized with    D.   setosum    by  Mortensen (1940) . The holotype of    D.   clarki    has not been found, but three unlabeled dried tests of    Diadema    were found among Ikeda’s original collection held in the Kitakyushu Museum of Natural History and Human History, Fukuoka, Japan. A short mtDNA  COI  fragment (ca. 350bp) was amplified from one of the tests, and the nucleotide sequence determined (275bp) was nearly identical with that of    Diadema   -sp. Arrangements of the primary tubercles on the coronal plates in    Diadema   -sp and the museum specimen also conformed with    D.   clarki   , indicating that    Diadema   -sp is identical to    D.   clarki    and a valid species. Narrow latitudinal distribution (31°N to 35°N) of    D.   clarki    in Japan was observed, where it co-existed with abundant    D.   setosum    and rare    D.   savignyi   . No    D.   clarki    was found in the southern islands in Japan, such as Satsunan Islands to Ryukyu Islands and Ogasawara Island, where    D.   setosum    and    D.   savignyi    were commonly observed.

DNA barcoding and morphological analyses revealed validity of Diadema clarki Ikeda, 1939 ( Echinodermata , Echinoidea , Diadematidae )