Complete mitochondrial genomes are valuable resources for different research fields such as genomics, molecular evolution and phylogenetics. The subfamily Lachninae represents one of the most ancient evolutionary lineages of aphids. To date, however, no complete Lachninae mitogenome is available in public databases. Here we report the Stomaphis sinisalicis mitogenome, representing the first complete mitogenome of Lachninae. The S. sinisalicis mitogenome is consist of 13 protein-coding genes (PCGs), two rRNA genes (rRNAs), 22 tRNA genes (tRNAs), a control region and a large tandem repeat region. Strikingly, the mitogenome exhibits a novel, highly rearranged gene order between trnE and nad1 compared with that of other aphids. The presence of repeat region in the basal Lachninae may further indicate it is probably an ancestral feature of aphid mitogenomes. Collectively, this study provides new insights on mitogenome evolution and valuable data for future comparative studies across different insect lineages.

The complete mitochondrial genomes and their rearrangement patterns can provide useful information for inferring evolutionary history of organisms. Aphids are one of the insect groups with some unique mitogenome features. In this study, to examine whether some features in aphid mitogenomes are independent species-specific evolutionary events or clade-specific events at certain taxonomic levels, we sequenced three new aphid mitogenomes (Hormaphidinae: Ceratovacuna keduensis, Pseudoregma panicola; Lachninae: Nippolachnus piri) and compared them with all known aphid mitogenomes. The three mitogenomes are 16,059–17,033 bp in length, with a set of 37 typical mitochondrial genes, a non-coding control region and a tandem repeat region. The gene orders of them are all highly rearranged. Within the subfamily Hormaphidinae, the presence of repeat region and mitogenome rearrangement in Cerataphidini species but not in the other two tribes indicate that these may be Cerataphidini-specific features. The same gene rearrangement pattern in the two Lachninae species, N. piri (Tuberolachnini) and Stomaphis sinisalicis (Stomaphidini), supports that this feature should be at least derived from the common ancestor of two tribes. Overall, our data and analyses provide new insights into the evolutionary patterns of gene rearrangement and repeat region in aphid mitogenomes, and further corroborate the potential role of gene rearrangement in elucidating the evolutionary history of different insect lineages.

Aphids are associated with an array of symbionts that have diverse ecological and evolutionary effects on their hosts. To date, symbiont communities of most aphid species are still poorly characterized, especially for the social aphids. In this study, high-throughput 16S rDNA amplicon sequencing was used to assess the bacterial communities of the social aphid Pseudoregma bambucicola, and the differences in bacterial diversity with respect to ant attendance and time series were also assessed. We found that the diversity of symbionts in P. bambucicola was low and three dominant symbionts (Buchnera, Pectobacterium and Wolbachia) were stably coexisting. Pectobacterium may help P. bambucicola feed on the hard bamboo stems, and genetic distance analysis suggests that the Pectobacterium in P. bambucicola may be a new symbiont species. Wolbachia may be associated with the transition of reproduction mode or has a nutritional role in P. bambucicola. Statistical tests on the diversity of bacterial communities in P. bambucicola suggest that aphid populations attended by ants usually have a significantly higher evenness than populations without ant attendance but there was no significant difference among aphid populations from different seasons.

Cytochrome P450 monooxygenases (P450s) have a variety of functions, including involvement in the metabolism of exogenous substances and the synthesis and degradation of endogenous substances, which are important for the growth and development of insects. Pseudoregma bambucicola is a social aphid that produces genetically identical but morphologically and behaviorally distinct first-instar soldiers and normal nymphs within colonies. In this study, we identified 43 P450 genes based on P. bambucicola genome data. Phylogenetic analysis showed that these genes were classified into 4 clans, 13 families, and 23 subfamilies. The CYP3 and CYP4 clans had a somewhat decreased number of genes. In addition, differential gene expression analysis based on transcriptome data showed that several P450 genes, including CYP18A1, CYP4G332, and CYP4G333, showed higher expression levels in soldiers compared to normal nymphs and adult aphids. These genes may be candidates for causing epidermal hardening and developmental arrest in soldiers. This study provides valuable data and lays the foundation for the study of functions of P450 genes in the social aphid P. bambucicola.

Social aphids produce different morphs, which are genetically identical but morphologically different. Each morph performs a different duty in its community. Social aphids usually produce morphologically distinct soldiers to protect their colonies. The social aphid Pseudoregma bambucicola produces sterile first instar soldiers with specialized body parts and unique defensive behaviors, such as hind leg waving. By using this species as a research model, this study tested the assumption that the functional morphological basis of defensive behaviors of soldiers is related to specialized body parts. Field observations and a comprehensive morphometric analysis were carried out for natural populations. The results showed significant differences in functional morphology between soldiers, first instar nymphs, and adults. Elongated hind legs in soldiers are an important functional morphological basis for the deimatic behavior of hind leg waving, while sclerotized front legs and head horns are related to the function of directly attacking natural enemies. The size variation of different body parts among different morphs also indicates a cost–benefit trade-off in the evolution of the social aphid species.

Species identification of Aphytis on the basis of adult morphology is extremely difficult, especially in the A. lingnanensis group, with several cryptic species. Pupal pigmentation could be used as one of the taxonomic characters for Aphytis species, and in some instances, pupal pigmentation actually provided the first clue to the distinctness of cryptic Aphytis species. The present study investigated the full-grown larvae or younger pupae of Aphytis species, and pupal pigmentation and pupal development were observed and photographed. Four characteristic pigmentation patterns of Aphytis pupae were summarized including: entirely yellow, partly dark brown, entirely or predominantly black, and partly black. The species in the chilensis and mytilaspidis groups, and some unassigned species, generally have entirely, or predominantly and or partly black pupae. The species in the chrysomphali, funicularis, and proclia groups generally have the pupae entirely yellow. The species of the lingnanensis group have the pupae both entirely yellow, e.g., A. fisheri, and partly dark brown pupae, e.g., A. lingnanensis, A. holoxanthus and A. melinus. The pupae of Aphytis species in this study had a developmental duration of about 5–8 days at 27 ± 1 °C, 70 ± 5% RH and a photoperiod of 10L: 14D. It was found that the pupal skin was always melanized at the beginning stage, generally in the first day, among the pigmented pupae of Aphytis species. As development continued, the pigmentation became darker and the eye colour changed from pale red/brown to green. No significant intra-specific variation in pupal colour pattern was detected despite relatively high numbers of specimens examined, many from multiple, and different, origins. Overall, our study indicates that pupal pigmentation could be more helpful in species identification of Aphytis.