Parastichopus regalis (Cuvier, 1817) is the most expensive seafood product on the Catalonian market (NE Spain), with prices at approximately 130 €/Kg (fresh weight). Despite its ecological and economic importance, biological and genetic information on this sea cucumber species is scarce. Here, we provide both the first insight on the genetic structure of P. regalis using sequences of cytochrome oxidase I (COI) and 16S genes and a morphological description of its population. Individual sea cucumbers were collected in six locations along the Spanish Mediterranean coast, including an area under fishery pressure (Catalonia). We found high haplotype diversity and low nucleotide diversity for both genes, with higher levels of genetic diversity observed in the COI gene. The population pairwise fixation index (F ST ), AMOVA and correspondence analysis (CA) based on the COI gene revealed significant genetic differentiation among some locations. However, further analysis using nuclear markers (e.g., microsatellites) is necessary to corroborate these results. Moreover, the genetic and morphological data may indicate fishery effects on the Catalonian population with a decrease in the size and weight averages and lower genetic diversity compared with locations that lack fishery pressure. For the appropriate management of this species, we suggest the following: 1) accurately assessing the stock status along the Spanish coasts; 2) studying the reproductive cycle of this target species and the establishment of a closed fishery season according to the reproductive cycle; and 3) establishing protected areas (i.e., not take zones) to conserve healthy populations and favour recruitment in the nearby areas.