Photoperiodism of Diapause Induction in the Silkworm, Bombyx mori.

The silkworm Bombyx mori exhibits a photoperiodic response (PR) for embryonic diapause induction. This article provides a comprehensive review of literature on the silkworm PR, starting from early works on population to recent studies uncovering the molecular mechanism. Makita Kogure (1933) conducted extensive research on the PR, presenting a pioneering paper on insect photoperiodism. In the 1970s and 80s, artificial diets were developed, and the influence of nutrition on PR was well documented. The photoperiodic photoreceptor has been investigated from organ to molecular level in the silkworm. Culture experiments demonstrated that the photoperiodic induction can be programmed in an isolated brain (Br)-subesophageal ganglion (SG) complex with corpora cardiaca (CC)-corpora allata (CA). The requirement of dietary vitamin A for PR suggests the involvement of opsin pigment in the photoperiodic reception, and a cDNA encoding an opsin (Boceropsin) was cloned from the brain. The effector system concerning the production and secretion of diapause hormone (DH) has also been extensively investigated in the silkworm. DH is produced in a pair of posterior cells of SG, transported to CC by nervi corporis cardiaci, and ultimately released into the hemolymph. Possible involvement of GABAergic and corazonin (Crz) signal pathways was suggested in the control of DH secretion. Knockout (KO) experiments of GABA transporter (GAT) and circadian clock genes demonstrated that GAT plays a crucial role in PR through circadian control. A model outlining the PR mechanism, from maternal photoperiodic light reception to DH secretion, has been proposed.

Fat enough for the winter? Does nutritional status affect diapause?

Many insects enter a dormant state termed diapause in anticipation of seasonal inhospitable conditions. Insects drastically reduce their feeding during diapause. Their reduced nutrient intake is paired with substantial nutrient costs: maintaining basal metabolism during diapause, repairing tissues damaged by adverse conditions, and resuming development after diapause. Many investigators have asked "Does nutrition affect diapause?" In this review, we survey the studies that have attempted to address this question. We propose the term nutritional status, a holistic view of nutrition that explicitly includes the perception, intake, and storage of the great breadth of nutrients. We examine the studies that have sought to test if nutrition affects diapause, trying to identify specific facets of nutritional status that affect diapause phenotypes. Curiously, low quality host plants during the diapause induction phase generally induce diapause, but food deprivation during the same phase generally averts diapause. Using the geometric framework of nutrition to identify specific dietary components that affect diapause may reconcile these contrasting findings. This framework can establish nutritionally permissive space, distinguishing nutrient changes that affect diapause from changes that induce other dormancies. Refeeding is another important experimental technique that distinguishes between diapause and quiescence, a non-diapause dormancy. We also find insufficient evidence for the hypothesis that nutrient stores regulate diapause length and suggest manipulations to investigate the role of nutrient stores in diapause termination. Finally, we propose mechanisms that could interface nutritional status with the diapause program, focusing on combined action of the nutritional axis between the gut, fat body, and brain.

Global Transcriptome Sequencing Reveals Molecular Profiles of Summer Diapause Induction Stage of Onion Maggot, Delia antiqua (Diptera: Anthomyiidae).

The onion maggot, Delia antiqua, is a worldwide subterranean pest and can enter diapause during the summer and winter seasons. The molecular regulation of the ontogenesis transition remains largely unknown. Here we used high-throughput RNA sequencing to identify candidate genes and processes linked to summer diapause (SD) induction by comparing the transcriptome differences between the most sensitive larval developmental stage of SD and nondiapause (ND). Nine pairwise comparisons were performed, and significantly differentially regulated transcripts were identified. Several functional terms related to lipid, carbohydrate, and energy metabolism, environmental adaption, immune response, and aging were enriched during the most sensitive SD induction period. A subset of genes, including circadian clock genes, were expressed differentially under diapause induction conditions, and there was much more variation in the most sensitive period of ND- than SD-destined larvae. These expression variations probably resulted in a deep restructuring of metabolic pathways. Potential regulatory elements of SD induction including genes related to lipid, carbohydrate, energy metabolism, and environmental adaption. Collectively, our results suggest the circadian clock is one of the key drivers for integrating environmental signals into the SD induction. Our transcriptome analysis provides insight into the fundamental role of the circadian clock in SD induction in this important model insect species, and contributes to the in-depth elucidation of the molecular regulation mechanism of insect diapause induction.

The opening-closing rhythms of the subelytral cavity associated with gas exchange patterns in diapausing Colorado potato beetle, Leptinotarsa decemlineata.

The opening-closing rhythms of the subelytral cavity and associated gas exchange patterns were monitored in diapausing Leptinotarsa decemlineata beetles. Measurements were made by means of a flow-through CO2 analyser and a coulometric respirometer. Under the elytra of these beetles there is a more or less tightly enclosed space, the subelytral cavity (SEC). When the cavity was tightly closed, air pressure inside was sub-atmospheric, as a result of oxygen uptake into the tracheae by the beetle. In about half of the beetles, regular opening-closing rhythms of the SEC were observed visually and also recorded; these beetles displayed a discontinuous gas exchange pattern. The SEC opened at the start of the CO2 burst and was immediately closed. On opening, a rapid passive suction inflow of atmospheric air into the SEC occurred, recorded coulometrically as a sharp upward peak. As the CO2 burst lasted beyond the closure of the SEC, we suggest that most of the CO2 was expelled through the mesothoracic spiracles. In the remaining beetles, the SEC was continually semi-open, and cyclic gas exchange was exhibited. The locking mechanisms and structures between the elytra and between the elytra and the body were examined under a stereomicroscope and by means of microphotography. We conclude that at least some of the L. decemlineata diapausing beetles were able to close their subelytral cavity tightly, and that the cavity then served as a water-saving device.

Cloning of cDNA encoding a Bombyx mori homolog of human oxidation resistance 1 (OXR1) protein from diapause eggs, and analyses of its expression and function.

To better understand the molecular mechanisms of diapause initiation, we used the sensitive cDNA subtraction (selective amplification via biotin- and restriction-mediated enrichment) method and isolated a novel gene expressed abundantly in diapause eggs of the silkworm, Bombyx mori, which encodes a homolog of the human oxidation resistance 1 (OXR1) protein. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting analyses confirmed that BmOXR1 mRNA and its 140-kDa protein were differentially expressed in diapause eggs compared to non-diapause eggs. OXR1 double-stranded RNA (dsRNA) was injected into diapause-destined eggs before the cellular blastoderm stage, and 4days later, when untreated eggs reached the diapause stage, the OXR1 protein disappeared; however, these eggs remained in diapause, suggesting that BmOXR1 is not essential for diapause initiation and/or maintenance. To further investigate the in vivo function of BmOXR1 apart from its role in diapause, we overexpressed BmOXR1 in Drosophila melanogaster. The fruit fly male adult life-span was significantly extended in the 50%-survival time when adults were reared on diets both with and without H2O2 solution under 25°C incubation. These results suggest that BmOXR1 functions in D. melanogaster via a possible antioxidant effect. As BmOXR1 was expressed mainly in the nuclei of D. melanogaster cells, the mechanism underlying its antioxidation effect appears to be different from that in humans where it is expressed mainly in the mitochondria. Taken together, these results suggest that BmOXR1 might serve as an antioxidant regulator during the early diapause stage.

Identification of a putative antifreeze protein gene that is highly expressed during preparation for winter in the sunn pest, Eurygaster maura.

A cDNA library generated from the fat body of field-collected, diapausing adults of the sunn pest, Eurygaster maura revealed the presence of a transcript that encodes a protein that shares the distinct physiochemical and structural features of an insect antifreeze protein. The transcript, which is most abundant in the midgut, accumulates in adults as they leave the fields in late summer and migrate to surrounding mountainous areas to overwinter. Transcript abundance again declines when adults return to the fields the following spring. This winter pattern of abundance suggests that this protein may be critical for winter survival in the cold regions where the bug enters its obligatory diapause.