DNA barcoding has become a promising means for the identification of organisms of all life-history stages. Currently, distance-based and tree-based methods are most widely used to define species boundaries and uncover cryptic species. However, there is no universal threshold of genetic distance values that can be used to distinguish taxonomic groups. Alternatively, DNA barcoding can deploy a “character-based” method, whereby species are identified through the discrete nucleotide substitutions. Our research focuses on the delimitation of moth species using DNA-barcoding methods. We analyzed 393 Lepidopteran specimens belonging to 80 morphologically recognized species with a standard cytochrome c oxidase subunit I (COI) sequencing approach, and deployed tree-based, distance-based, and diagnostic character-based methods to identify the taxa. The tree-based method divided the 393 specimens into 79 taxa (species), and the distance-based method divided them into 84 taxa (species). Although the diagnostic character-based method found only 39 so-identifiable species in the 80 species, with a reduction in sample size the accuracy rate substantially improved. For example, in the Arctiidae subset, all 12 species had diagnostics characteristics. Compared with traditional morphological method, molecular taxonomy performed well. All three methods enable the rapid delimitation of species, although they have different characteristics and different strengths. The tree-based and distance-based methods can be used for accurate species identification and biodiversity studies in large data sets, while the character-based method performs well in small data sets and can also be used as the foundation of species-specific biochips.

A new subgenus, Reticularisus Wu, Wu & Han, subgen. n., with type species Rhamnosa henanensis Wu, 2008, of the genus Rhamnosa Fixsen, 1887 is described and illustrated. For the sake of contrast, the type species of the other two subgenera in this genus have been given, including adults and male genitalia. Rhamnosa (Reticularisus) shierbeihoua Wu, Wu & Han, sp. n., a Limacodidae collected from the southwest of China is described as new to science. Also, the new species is illustrated with images of the adult and male genitalia and compared with the similar species Rh. henanensis.

Four new species, Kitanola shilinensissp. nov., K. elegantasp. nov., Fignya ravalbasp. nov., and Euphlyctinides pseudolaikasp. nov., are described from southwestern China. Two species are reported new to China, Euphlyctinidis indi Solovyev, 2009 and Limacocera pachycera (Hampson, 1897). The adults and genitalia of all the treated species are illustrated. A checklist for the species belonging to the treated genera is provided.

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The discovery of Striogyia simianshanasp. nov. (Lepidoptera: Limacodidae) resulted from a research project investigating the biodiversity of the Simian Mountain Nature Reserve in Chongqing, Southwest China. This new species is characterised by several distinct features, namely the black, broken postmedial line of the forewing and the long filiform antenna found in males. The shape of its valva, transtilla, juxta and phallus also distinguish it from other congeners. Furthermore, images of the adult habitus and male genitalia are compared with similar species S. obatera Wu, 2011 and S. acuta Wu, 2020, which further illustrate characteristics unique to S. simianshana. A distribution map and key for Chinese Striogyia species are provided.