Phylogenetic implications based on an ultrastructural study with emphasis on the tracheal system of the compound eyes of three species of nymphalid butterflies.

Compound eyes are the prominent visual organs of insects and can provide valuable information for the reconstruction of insect phylogeny. Although the largest butterfly family (Nymphalidae) has been well defined, the infrafamilial phylogenetic relationships remain controversial hitherto. In the present study the ultrastructure of the compound eyes of three nymphalids Neptis beroe, Childrena zenobia, and Palaeonympha opalina was investigated using light and transmission electron microscopy in an attempt to seek potentially important phylogenetic characters. The compound eyes of the nymphalids share a tracheal system in a "1-4-8" branching pattern. The eight tracheal subbranches exhibit distinct distribution patterns along the basal retinula cell as follows: the tracheal subbranches of Palaeonympha opaline are close to the rhabdom in the distance from the distalmost part of the basal retinula cell to the rhabdom end, while those of N. beroe and C. zenobia are on the periphery of the retinula along almost the whole basal retinula cell and become close to the rhabdom just at the proximal end of the basal retinula cell. The tracheal structure of the three nymphalids is discussed for their potential phylogenetic implications.

Ultrastructure of Ejaculatory Ducts of Cerapanorpa nanwutaina and Furcatopanorpa longihypovalva (Mecoptera: Panorpidae).

The ultrastructure of the ejaculatory duct was investigated in the scorpionflies Cerapanorpa nanwutaina (Chou 1981) and Furcatopanorpa longihypovalva (Hua & Cai, 2009) (Mecoptera: Panorpidae) using light and transmission electron microscopy. The ejaculatory ducts of both species comprise a median duct and an accessory sac. The median duct consists of a basal lamina, a mono-layered epithelium, a subcuticular cavity, and an inner cuticle. The accessory sac contains a single layer of epithelium and a basal lamina. A muscular layer is present in the accessory sac of C. nanwutaina and in the median duct of F. longihypovalva. The epithelia in the median duct and the accessory sac are well developed, their cells containing numerous cisterns of rough endoplasmic reticulum, mitochondria, and microvilli. The secretions of the median duct are first extruded into the subcuticular cavity and then into the lumen through an inner cuticle, while the secretions of the accessory sac are discharged directly into the lumen. The ejaculatory duct of F. longihypovalva is longer and has thicker epithelium with more cell organelles and secretions than that of C. nanwutaina.

Identification and expression profiles of candidate chemosensory receptors in Histia rhodope (Lepidoptera: Zygaenidae).

Insect olfaction and vision play important roles in survival and reproduction. Diurnal butterflies mainly rely on visual cues whereas nocturnal moths rely on olfactory signals to locate external resources. Histia rhodope Cramer (Lepidoptera: Zygaenidae) is an important pest of the landscape tree Bischofia polycarpa in China and other Southeast Asian regions. As a diurnal moth, H. rhodope represents a suitable model for studying the evolutionary shift from olfactory to visual communication. However, only a few chemosensory soluble proteins have been characterized and information on H. rhodope chemoreceptor genes is currently lacking. In this study, we identified 45 odorant receptors (ORs), nine ionotropic receptors (IRs), eight gustatory receptors (GRs) and two sensory neuron membrane proteins (SNMPs) from our previously acquired H. rhodope antennal transcriptomic data. The number of chemoreceptors of H. rhodope was less compared with that found in many nocturnal moths. Some specific chemoreceptors such as OR co-receptor (ORco), ionotropic receptors co-receptor, CO2 receptors, sugar receptors and bitter receptors were predicted by phylogenetic analysis. Notably, two candidate pheromone receptors (PRs) were identified within a novel PR lineage. qRT-PCR results showed that almost all tested genes (22/24) were predominantly expressed in antennae, indicating that they may be important in olfactory function. Among these antennae-enriched genes, six ORs, five IRs and two GRs displayed female-biased expression, while two ORs displayed male-biased expression. Additionally, HrhoIR75q.2 and HrhoGR67 were more highly expressed in heads and legs. This study enriches the olfactory gene inventory of H. rhodope and provides the foundation for further research of the chemoreception mechanism in diurnal moths.

Ultrastructure of the vasa deferentia of Terrobittacus implicatus and Cerapanorpa nanwutaina (Insecta: Mecoptera).

The fine structures of vasa deferentia and postvesicular vasa deferentia were investigated in the hangingfly Terrobittacus implicatus (Cai et al. 2006) and the scorpionfly Cerapanorpa nanwutaina (Chou 1981) using light and transmission electron microscopy, and schematic diagrams were drawn accordingly. The vasa deferentia of both species comprise muscular layers, a basal lamina, and a mono-layered epithelium, but the postvesicular vasa deferentia contain muscular layers, a basal lamina, a single-layered epithelium, a subcuticular cavity, and an inner cuticle respectively. The vas deferens releases secretions into the lumen directly, probably by means of merocrine production. On the contrary, the cells of the postvesicular vas deferens correspond to class I glandular cells, discharging secretions into the subcuticular cavity first, and then into the lumen through an inner cuticle. The epithelium in both structures of Bittacidae is well developed and contains more microvilli, organelles, and more types of secretions than in Panorpidae. In Panorpidae, the spine of the postvesicular vas deferens may serve as a barricade for the reflow of the sperm and to protect the extraordinarily long structure from being collapsed or injured.

Ultrastructure and function of the seminal vesicle of Bittacidae (Insecta: Mecoptera).

The fine structure of the seminal vesicle and reproductive accessory glands was investigated in Bittacidae of Mecoptera using light and transmission electron microscopy. The male reproductive system of Bittacidae mainly consists of a pair of testes, a pair of vasa deferentia, and an ejaculatory sac. The vas deferens is greatly expanded for its middle and medio-posterior parts to form a well-developed seminal vesicle. The seminal vesicle is composed of layers of developed muscles and a mono-layered epithelium surrounding the small central lumen. The epithelium is rich in rough endoplasmic reticulum and mitochondria, and secretes vesicles and granules into the central lumen by merocrine mechanisms. A pair of elongate mesodermal accessory glands opens into the lateral side of the seminal vesicles. The accessory glands are similar to the seminal vesicle in structure, also consisting of layers of muscle fibres and a mono-layered elongated epithelium, the cells of which contain numerous cisterns of rough endoplasmic reticulum and mitochondria, and a few Golgi complexes. The epithelial cells of accessory glands extrude secretions via apocrine and merocrine processes. The seminal vesicles mainly serve the function of secretion rather than temporarily storing spermatozoa. The sperm instead are temporarily stored in the epididymis, the greatly coiled distal portion of the vas deferens.

Ultrastructure of male accessory glands in the scorpionfly Sinopanorpa tincta (Navás, 1931) (Mecoptera: Panorpidae).

The ultrastructure of male reproductive accessory glands was investigated in the scorpionfly Sinopanorpa tincta (Navás, 1931) (Mecoptera: Panorpidae) using light and transmission electron microscopy. The male accessory glands comprise one pair of mesodermal glands (mesadenia) and six pairs of ectodermal glands (ectadenia). The former opens into the vasa deferentia and the latter into the ejaculatory sac. The mesadenia consist of a mono-layered elongated columnar epithelium, the cells of which are highly microvillated and extrude secretory granules by means of merocrine mechanisms. The epithelium of ectadenia consists of two types of cells: the large secretory cells and the thin duct-forming cells. These two types of cells that join with a cuticular duct constitute a functional glandular unit, corresponding to the class III glandular cell type of Noirot and Quennedey. The cuticular duct consists of a receiving canal and a conducting canal. The secretory granules were taken up by the receiving canal and then plunged into the lumen through the conducting canal.