We recognize a new species, belonging to the genus Neopsephus Kishii 1990, from Is. Jeju. The novel species is closely allied to N. takasago Kishii 1990 known from Taiwan. This study provides the detailed morphological characteristics investigated by SEM and molecular data inferred from COI and 16S rRNA of mitochondria.

DNA barcoding is a strong species identification tool for all animal taxa, and can easily be conducted when materials are under DNA friendly conditions. In contract, a full-length (659 bp) sequencing has been limited for the degraded DNAs extracted from old museum specimens. The initial challenges to retrieve the authentic DNA fragments from old museum specimens were attempted by obtaining short sequences (<300 bp) with the cloning process after PCR, making it both expensive and time-consuming. In this study, we employed a modified method to analyze the full-length DNA barcoding regions in 31~52 year-old butterfly specimens (301 dried specimens of 39 species) using direct sequencing after PCR with two different methods: 1) the successful PCR rates of 0 to 5.6% using four universal primer sets were too low to obtain authentic sequences and the cross-contamination was detected in almost all successful amplicons; 2) the success rates of PCR using specie-specific overlapping primer sets were distinctly high, reaching up to 75% with 98% authentic and 2% non-specific sequences. Thus, the result showed the method that using species-specific primer set per species yields the most effective success rates of both PCR and sequencing from degraded DNA without incorrect sequences.

In traditional taxonomy on the family Cantharidae, color pattern of the body and the structure of the male genitalia have been often used as diagnostic characters in identification of the specific level. However, these characters caused the difficulty in identifying the female in case a species was described only by male specimens or has the several color types among individuals. In this study, we attempted to evaluate the species reality of Asiopodabrus fragiliformis which was often difficult to be identified due to individual variation in color pattern and lack of information of female, through searching for new morphological diagnostic characters as well as DNA barcoding analysis, including their closely relative species from Russia and Japan. The results showed that A. fragiliformis was represented as three clusters strongly supported by high value of boots trap (>99%) and over 3% branch length. The pairwise distances between species of Asiopodabrus were detected larger, ranged from 3.4–9.5%, than the intragroup distance ranged from 0–2.9% indicating presence of a barcoding gap. And then, the three clusters were respectively determined as A. fragiliformis, A. kurvatovi and a new species through the analysis of morphology and COI gene. Therefore, we suggest that the species delineation on polymorphic species and the female specimens of closely resembling species would be more exactly and effectively determined if DNA barcoding and the traditional taxonomy are used as complementary methods for identification.