To assess the accuracy of species delimitation in the genus Aphaenomurus Yosii, we conducted a comparative micro-morphological study and molecular analysis using two mitochondrial (COI, 16S) and two nuclear genes (18S, 28S) on 118 specimens from 24 localities. The results showed that the morphological characters of A. vicinus and A. interpositus, as presented in the original description, were intermixed in phylogenetic lineages within the genus and did not form independent lineages. Furthermore, there were enough differences among Aphaenomurus individuals to be considered as morphologically distinct species (Th. III is 1+1 or 2+2, Abd. I is 2+2 or 3+3, Abd. II is 2+2 or 3+3), but they do not form an independent lineage. Molecular and morphological analyses have determined that A. vicinus and A. interpositus are the same species, exhibiting morphological variation in dental spines formula, claws, chaetotaxy, and other features. Additionally, the genus Aphaenomurus forms a monophyletic clade, which is further divided into several lineages within the genus. No morphological differences were observed to distinguish these lineages. This cladistic divergence is attributed to heteroplasmy, which is supported by previous studies that have suggested the possibility and problems of heteroplasmy in Collembola, and by the high genetic distances between individuals in the mitochondrial genes of Aphaenomurus.

The coastal Aleochara fucicola Sharp species complex include A. fucicola Sharp, A. hayamai Yamamoto and Maruyama, A. puetzi (Assing), and A. segregata Yamamoto and Maruyama. To clarify the delineation and phylogenetic relationships of the A. fucicola species complex, we conducted phylogenetic analyses based on molecular characters and detailed morphological characters by using scanning electron microscopy (SEM) images. The dataset comprised two mitochondrial (cytochrome c oxidase I and COII) and three nuclear protein coding genes (CAD, EF1-a, and Wingless) for 34 populations of four species of A. fucicola species complex. We found strong disagreement between mitochondrial and nuclear gene trees. Two species, A. puetzi and A. segregata, were not distinguished by mitochondrial DNA data, but distinguished by nuclear DNA data and SEM characters. A multilocus phylogeny and SEM images revealed four well-supported species lineages. Based on a multicolus phylogeny and SEM images, we conclude the following the taxonomic status of the Aleochara fucicola species complex with four species (A. fucicola, A. hayamai, A. puetzi, and A. segregata).

Due to their ease of sequencing and high levels of nucleotide diversity mitochondrial (mt) genes have long been favoured targets for investigations of species level issues in systematic entomology particularly species delimitation and diagnostic identification. Advances in sequencing technology and price reductions have now made routine sequencing of whole insect mt genomes feasible and thus the application of much larger datasets to these perennial problems can be attempted. I will present two examples – the development of diagnostic targets within forensically important blowflies (Diptera: Calliphoridae) from scans of whole mt genome variability and the determination of species status within the cotton bollworm (Lepidoptera: Noctuiidae: Helicoverpa) – to illustrate the ease and comparative cheapness of genomics level approaches to these age old problems.