Unraveling the intricate biodiversity of the benthic harpacticoid genus ��it��Nannopus��/it�� (Copepoda, Harpacticoida, Nannopodidae) in Korean waters

Nannopus (Harpacticoida, Nannopodidae) species are abundant and widely distributed throughout the world across a variety of habitats. Nannopus is well known for high frequencies of misidentifications and thus may comprise several cryptic complexes and morphologically distinct species. Cryptic taxa are common in meiofauna communities. In this study, we aimed to identify Nannopus species using an integrative approach including molecular taxonomy. We adopted a non-destructive DNA extraction method so that morphological and molecular data could be obtained from the same specimen. We analyzed the molecular diversity and distributions of Nannopus using a total of 190 individuals. We sequenced the 190 mtCOI, 53 mtCYTB, 25 18SrDNA, and 4328SrDNA genes from 190 individuals. Several species delimitation approaches were applied, including uncorrected p-distances for mtCOI, mtCYTB, 18SrDNA, and 28SrDNA, and Automatic Barcode Gap Discovery and Bayesian implemented Poisson tree processes for mtCOI and mtCYTB data. The maximum likelihood and Bayesian approaches were used to examine the phylogenetic relationships among individuals using the combined set of all four genes. Our species delimitation and phylogenetic analyses indicated the presence of three cryptic and six morphologically distinct species. All species are sympatric and widely distributed across mudflats ranging from the Yellow Sea to the South Sea in Korea. The divergence patterns of the four genes were not congruent. A phylogenetic tree based on the concatenated dataset was the most robust, was congruent with morphology, and suggested two major clades. We considered the validity of reinstating the genus Ilyophilus (Lilljeborg, 1902) and ultimately concluded that including all congeners in Nannopus until the type species (N. palustris Brady, 1880) is re-described was the most prudent approach.We would like to thank Jeongho Kim for assisting in the collection of sediments from type localities. We are very grateful to Dr. Terue Cristina Kihara, and her institute (DZMB, Germany) for permitting us to use the photo of confocal image for graphical abstract. This study was supported by a grant entitled "Discovery project of endemic species in Korea" from the National Institute of Biological Resources funded by the Ministry of Environment, Korea (NIBR 201601201), and a grant (2018R1D1A1B07050117) from Korea Research Foundation, and the BK21 Plus Program (Eco-Bio Fusion Research Team, 22A20130012352) funded by the Ministry of Education (MOE, Korea) to WL. This work was additionally supported by National Research Foundation of Korea Grant (2018R1C1B3001650) and the Chung-Ang University Research Grant in 2017 to SE