Integrative taxonomy on the rare sky-island Ligidium species from southwest China (Isopoda, Oniscidea, Ligiidae)

The sky-island Ligidium species fauna in southwest China is poorly known. Before this study, six of the seven sky-island species of the genus were known to be endemic to southwest China. In morphology, Ligidium species are often difficult to identify, and an appraisal of integrative taxonomy is needed. We integrated morphology and molecular analyses to delimit Ligidium species. Molecular species delimitation based on distance- and evolutionary models recovered seven-candidate lineages from five gene markers (COI, 12S rRNA, 18S rRNA, 28S rRNA and NAK). We also estimated that the species divergences of sky-island Ligidium in southwest China started in late Eocene (40.97 Mya) to middle Miocene (15.19 Mya). Four new species (L. duospinatum Li, sp. nov., L. acuminatum Li, sp. nov., L. rotundum Li, sp. nov. and L. tridentatum Li, sp. nov.) are described. Morphological confusion among L. denticulatum Shen, 1949, L. inerme Nunomura & Xie, 2000 and L. sichuanense Nunomura, 2002 is clarified by integrative taxonomy. This work confirms that an integrative approach to Ligidium taxonomy is fundamental for objective classification, and deduced the uplift of Qinghai-Tibetan Plateau in the late Eocene and middle Miocene as one of the principal reasons for the species divergences of sky-island Ligidium in southwest China. We also inferred that sky-island mountains have a huge reserve of higher Ligidium species diversity.