Four complete mitogenomes of Eysarcoris rosaceus, E. montivagus, E. gibbosus, E. annamita and one near-complete mitochondrial genome of E. ventralis were sequenced and used to explore the phylogenetic relationships of tribes within the subfamily Pentatominae. The mitogenomes range from 15,422 to 16,043 base pairs (bp) in length and encode 37 genes, including 13 protein-coding genes (PCGs), two rRNA genes, 22 tRNA genes (21 in E. ventralis), and a control region. Similar to other heteropteran species, the AT contents of the sequenced species were higher than their GC contents. The most frequently used start/stop codon was ATN/TAA. GTG was only found in atp6 and atp8 of E. gibbosus. All transfer RNA genes (tRNAs) exhibit the typical cloverleaf secondary structure, except for the trnS1 and trnV, which lacks the stem of the DHU arm. The length and copy number of repeat units were conserved within Eysarcoris, with the exception of E. gibbosus. Phylogenetic analyses based on mitogenomes using both maximum likelihood (ML) and Bayesian inference (BI) methods strongly supported the relationship among tribes within Pentatominae and confirmed that Graphosoma should be an intermediate lineage of Pentatominae. The relationship between Eysarcoris and Carbula was strongly supported and combined with our previous geometric morphometrics and chromosomal studies, suggest the Eysarcoris should belong to the tribe Eyasrcorini. This work will help to enhance our understanding of mitochondrial genomic evolution and phylogenetic relationships in Pentatominae.

Stag beetles (Coleoptera, Scarabaeoidea, Lucanidae) have received extensive attention from researchers in behavioral ecology and evolutionary biology. There have been no previous quantitative analyses, particularly using a geometric morphometric approach based on a large sample of data, to shed light on the morphological diversity and evolution of Lucanidae. Thoracic adaptation and ecological differentiation are intimately related, and the pronotum bears important muscles and supports the locomotion of prothoracic legs. The elytron is an autapomorphy of the Coleoptera. To reconstruct and visualize the patterns of evolutionary diversification and phylogenetic history of shape change, an ancestral groundplan can be reconstructed by mapping geometric morphometric data onto a phylogenetic tree. In this study, the morphologies of the pronotum and elytron in 1303 stag beetles (Lucanidae), including approximately 99.2% of all globally described species, were examined, thus revealing several aspects of morphological diversity and evolution. First, on the basis of geometric morphometric analysis, we found significant morphological differences in the pronotum or elytron between any two Lucanidae subfamilies. And we subsequently reconstructed the ancestral groundplans of the two structures in stag beetles and compared them with those of extant species (through cladistic and geometric morphometric methods). The ancestral groundplan of Lucanidae was found to be most similar to extant Nicagini in both the pronotum and elytron, according to Mahalanobis distances. Furthermore, we analyzed species richness and morphological diversity of stag beetles and the relationships between them and found that the two parameters were not always correlated. Aesalinae was found to be the most diverse subfamily in both the pronotum and elytron, despite its poor species richness, and the diversity of the pronotum or elytron was not superior in Lucaninae, despite its high species richness. Our study provides insights into the morphological variations and evolutionary history of the pronotum and elytron in four subfamilies of stag beetles, and it illuminates the relationship between morphological diversity and species richness. Intriguingly, our analysis indicates that morphological diversity and species richness are not always correlated. These findings may stimulate further studies in this field.

The astonishing spectrum of scarabaeine lifestyles makes them an attractive group for studies in entomology and evolutionary biology. As a result of adaptions to specific food substrates and textures, the mouthparts of dung beetles, particularly the mandible, have undergone considerable evolutionary changes and differ distinctly from the presumptive ancestral conditions of the Coleoptera and Polyphaga. The possible functions of dung beetle mouthparts and the evolution of dung feeding have been controversial for decades. In this study, 187 scarabs representing all tribes of the Scarabaeinae and the major lineages within the Scarabaeoidea, along with three major feeding types within the Scarabaeoidea (omnivory, phytophagy and coprophagy), were studied. Based on geometric morphometric and three-dimensional (3D) reconstruction approaches, morphological differences in mandibles among the three feeding types were identified. The ancestral forms of the mandible within the Scarabaeinae were reconstructed and compared with those of modern species. The most recent common ancestor of the Scarabaeinae fed on soft materials, and the ancestor of the Scarabaeinae and the Aphodiinae was in an evolutionary transition between processing more solid and softer substrates. Coprophagy originated from omnivorous ancestors that were very likely saprophagous. Furthermore, phytophagy may also have originated from omnivory. In addition, our study addresses the integration and modularity of geometric morphometric data in a phylogenetic context.

High-resolution 3D imaging technology has found a number of applications in many biological fields. However, the existing 3D imaging tools are often too time-consuming to use on large-scale specimens, such as centimeter-sized insects. In addition, most 3D imaging systems discard the natural color information of the specimens. To surmount these limitations, we present a structured illumination-based approach capable of delivering large field-of-view three-dimensional images. With this approach, 580nm lateral resolution full-color 3D images and 3D morphological data in the size range of typical insect samples can be obtained. This method provides a promising approach that can be used to support many different types of entomological investigations, including taxonomy, evolution, bionics, developmental biology, functional morphology, paleontology, forestry, etc.