Distinct roles of the Hox genes Ultrabithorax and abdominal-A in scorpionfly embryonic proleg development.

The abdominal appendages of larval insects have a complex evolutionary history of gain and loss, but the regulatory mechanisms underlying the abdominal appendage development remain largely unclear. Here, we investigated the embryogenesis of abdominal prolegs in the scorpionfly Panorpa liui Hua (Mecoptera: Panorpidae) using in situ hybridization and parental RNA interference. The results show that RNAi-mediated knockdown of Ultrabithorax (Ubx) led to a homeotic transformation of the first abdominal segment (A1) into the third thoracic segment (T3) and changed the distributions of the downstream target Distal-less (Dll) expression but did not affect the expression levels of Dll. Knockdown of abdominal-A (abd-A) resulted in malformed segments, abnormal prolegs and disrupted Dll expression. The results demonstrate that the gene Ubx maintains an ancestral role of modulating A1 appendage fate without preventing Dll initiation, and a secondary adaptation of abd-A evolves the ability to specify abdominal segments and proleg identity. We conclude that changes in abdominal Hox gene expression and their target genes regulate abdominal appendage morphology during the evolutionary course of holometabolous larvae.

The Hox gene Abdominal-B regulates the appendage development during the embryogenesis of scorpionflies.

The Homeotic Complex (Hox) genes encode conserved homeodomain transcription factors that specify segment identity and appendage morphology along the antero-posterior axis in bilaterian animals. The Hox gene Abdominal-B (Abd-B) is mainly expressed in the posterior segments of the abdomen and plays an important role in insect organogenesis. In Mecoptera, the potential function of this gene remains unclear yet. Here, we performed a de novo transcriptome assembly and identified an Abd-B ortholog in the scorpionfly Panorpa liui. Quantitative real-time reverse transcription PCR showed that Abd-B expression increased gradually in embryos 76 h post oviposition, and was mainly present in the more posterior abdominal segments. Embryonic RNA interference of Abd-B resulted in a set of abnormalities, including developmental arrest, malformed suckers and misspecification of posterior segment identity. These results suggest that Abd-B is required for the proper development of the posterior abdomen. Furthermore, in Abd-B RNAi embryos, the expression of the appendage marker Distal-less (Dll) was up-regulated and was additionally present on abdominal segments IX and X compared with wild embryos, suggesting that scorpionfly Abd-B may act to suppress proleg development and has gained the ability to repress Dll expression on the more posterior abdominal segments. This study provides additional information on both the functional and evolutionary roles of Abd-B across different insects.

Spermiogenesis of the hangingfly Terrobittacus implicatus (Huang and Hua) (Mecoptera: Bittacidae).

The structure of spermatozoa is able to provide valuable characters in resolving phylogenic relationships in Metazoa, especially in insects. Such data, however, are greatly deficient in Mecoptera. Here, we studied the spermiogenesis and ultrastructure of sperm in the hangingfly Terrobittacus implicatus (Huang and Hua) using transmission electron microscopy. The results show that the spermatogenesis of T. implicatus occurs within sperm cysts, following a pattern commonly found in insects. The microtubular doublets of spermatid axoneme exhibit a hooklike projection from the B-subtubule in the early period, but the projection disappears in the mature stage. The mature spermatozoon of T. implicatus is a filiform cell that is pronouncedly elongated and has a bi-layered acrosome, a nucleus with two lateral longitudinal grooves, a neck region with the centriole adjunct, a flagellum with a simple 9 + 2 axoneme, two extra-axonemal accessory structures, two accessory bodies, and two mitochondrial derivatives of unequal size, and a prominent glycocalyx. The basic structure of spermatozoa of T. implicatus is similar to that of other Mecoptera studied. However, this species shows characteristics unique in Bittacidae, such as the reniform appearance of the centriole adjunct, two triangular accessory bodies with granular materials, and two asymmetric mitochondrial derivatives with a circular profile in cross-section. The potential utilization of the sperm ultrastructure for understanding the phylogeny of Bittacidae is briefly discussed.