Pre-diapause Stages Research Articles

Lipid composition differs in diapause and nondiapause states of spotted stem borer, Chilo partellus

Spotted stem borer, Chilo partellus, undergoes larval diapause (hibernation and aestivation), and depends on the food reserve accumulated during feeding stage for its survival. Lipids are the primary source of energy during diapause, and essential for different cellular, biochemical and physiological functions. However, there is no information on lipid and lipophilic compound contents during different stages of hibernation, aestivation and nondiapause in C. partellus. Thus, we compared the concentration and composition of lipids in pre-diapause, diapause and post-diapause stages of hibernation and aestivation with nondiapause stages of C. partellus. The studies revealed significant differences in total lipids and various lipophilic compounds during different stages of diapause as compared to nondiapause C. partellus. The total lipids were significantly lower during diapause stage of aestivation and hibernation as compared to nondiapause larvae. Further, the linoleic acid, Methyl 3-methoxytetradecanoate, and l-(+)-Ascorbic acid 2,6-dihexadecanoate were significantly lower, and oleic and palmitoleic acids greater during pre-diapause and diapause stages of hibernation and aestivation as compared to nondiapause larvae. The cholesterol content was significantly greater during pre-diapause stage of hibernation, and diapause and post-diapause stages of aestivation as compared to nondiapause stages. The unsaturation ratio was significantly higher in the pre-diapause and diapause stages and lower in post-diapause stage of aestivation than the hibernation and nondiapause states. This study provides insights on differential lipid profiles during different phases of diapause, which could be useful for further understanding biochemical and physiological cross-talk, and develop target-specific technologies for the management of C. partellus.

Comparative 4D Label-Free Quantitative Proteomic Analysis of Bombus terrestris Provides Insights into Proteins and Processes Associated with Diapause.

Diapause, an adaptative strategy for survival under harsh conditions, is a dynamic multi-stage process. Bombus terrestris, an important agricultural pollinator, is declining in the wild, but artificial breeding is possible by imitating natural conditions. Mated queen bees enter reproductive diapause in winter and recover in spring, but the regulatory mechanisms remain unclear. Herein, we conducted a comparative 4D label-free proteomic analysis of queen bees during artificial breeding at seven timepoints, including pre-diapause, diapause, and post-diapause stages. Through bioinformatics analysis of proteomic and detection of substance content changes, our results found that, during pre-diapause stages, queen bees had active mitochondria with high levels of oxidative phosphorylation, high body weight, and glycogen and TAG content, all of which support energy consumption during subsequent diapause. During diapause stages, body weight and water content were decreased but glycerol increased, contributing to cold resistance. Dopamine content, immune defense, and protein phosphorylation were elevated, while fat metabolism, protein export, cell communication, signal transduction, and hydrolase activity decreased. Following diapause termination, JH titer, water, fatty acid, and pyruvate levels increased, catabolism, synaptic transmission, and insulin signaling were stimulated, ribosome and cell cycle proteins were upregulated, and cell proliferation was accelerated. Meanwhile, TAG and glycogen content decreased, and ovaries gradually developed. These findings illuminate changes occurring in queen bees at different diapause stages during commercial production.

Regulation of Juvenile Hormone on Summer Diapause of Geleruca daurica and Its Pathway Analysis.

Simple SummaryDiapause is an arrestment state in development, and plays an important role in life history in insects. It has been thought that a lack in juvenile hormone (JH) results in reproductive diapause occurring at the adult stage. However, we do not fully know about the underlying molecular mechanism. In this study, we proved that the topical application of a JH analog methoprene caused the changes at the transcriptional levels of a great number of genes, inhibited lipid accumulation, and finally delayed the adults entering diapause. Therefore, JH signaling plays an important role in regulating reproductive diapause of G. daurica, a new pest with great outbreaks in Inner Mongolia.Juvenile hormone (JH) signaling plays an important role in regulation of reproductive diapause in insects. However, we have little understanding of the effect of JH on gene expression at the transcriptome level in diapause. Galeruca daurica is a new pest in the Inner Mongolia grasslands with obligatory summer diapause in the adult stage. Topical application of a JH analog methoprene at the pre-diapause stage delayed the adults entering diapause and inhibited lipid accumulation whereas it did not during diapause. Using Illumina sequencing technology and bioinformatics tools, 54 and 138 differentially expressed genes (DEGs) were detected at 1 and 2 d after treatment, respectively. The KEGG analysis showed that the DEGs were mainly enriched in the metabolism pathways. qRT-PCR analysis indicated that methoprene promoted the expression of genes encoding vitellogenin, fork head transcription factor and Krüppel homolog 1, whereas suppressed the expression of genes encoding juvenile hormone-binding protein, juvenile hormone esterase, juvenile hormone acid methyltransferase, juvenile hormone epoxide hydrolase and fatty acid synthase 2. These results indicate that JH signaling plays an important role in regulating reproductive diapause of G. daurica.

Differences in the Development of Internal Reproductive Organs, Feeding Amount and Nutrient Storage between Pre-Diapause and Pre-Reproductive Harmonia axyridis Adults

Diapause control is one of the successful methods for long-term cold storage of biological control organisms without decreasing their fitness. Sufficient preparation before diapause is essential for successful diapause initiation and maintenance. Harmonia axyridis, an important biocontrol agent in its native place, can enter reproductive diapause for overwintering. However, the key preparatory events before diapause in H. axyridis, such as specific developmental trajectory, timing, and physiological changes, remain unclear. We compared differences in the development of internal reproductive organs, feeding amount and nutrient storage between pre-diapause and pre-reproductive adult H. axyridis which had been reared at 20 °C under a short-day (10L:14D) and long-day (14L:10D) condition, respectively. The results showed that there were obvious morphological differences in internal reproductive organs of diapause and reproductive H. axyridis. The development of internal reproductive organs was suppressed at early adult stage in pre-diapause beetles compared to pre-reproductive beetles. Feeding amount in both pre-diapause and pre-reproductive beetles increased for more than ten days after eclosion. The feeding amount of pre-diapause beetles sharply decreased from the 15th day after eclosion in females and the 14th day after eclosion in males, which implied the initiation of diapause. During the pre-diapause stage, carbohydrates and lipids were mainly accumulated by females, whereas males mainly accumulated carbohydrates. Our results not only provide basic information about the diapause process of pre-diapause stage, but also give tips to better utilization of diapause strategy during mass production and storage of H. axyridis.

The Prediapause Stage of Aedes japonicus japonicus and the Evolution of Embryonic Diapause in Aedini.

The genus Aedes is well known for its desiccation-resistant eggs, which frequently serve as an overwintering mechanism through diapause. Despite this, relatively little is known about the diapause and overwintering biology of most Aedes species including Aedes japonicus japonicus, an invasive mosquito in the United States. The importance of this mosquito in disease systems like La Crosse virus remain uncertain. Embryonic diapause is used by Ae. j. japonicus to survive temperate winters, and the persistence of this species in the Appalachian region is a result of overwintering, which has important implications for the transmission of this virus to humans. The objective of this study was to identify the prediapause stage, or the stage sensitive to environmental cues needed to induce diapause in this mosquito. By exposing each Ae. j. japonicus life stage independently to short-day photoperiods, we determined that the adult maternal life stage is the prediapause stage. Using the most recent phylogeny and prior literature on the prediapause stages in the genus Aedes, we were able to infer the evolutionary history of the prediapause stages of Aedes mosquitoes that overwinter or aestivate as eggs. This initial ancestral state reconstruction allowed us to hypothesize that Aedini mosquitoes that undergo obligate diapause may have evolved from those utilizing the embryonic prediapause stage, and that the ancestral prediapause state of Aedini appears to be maternally controlled.

Seasonal Synchronization of Diapause Phases in Aedes albopictus (Diptera: Culicidae).

In temperate areas, population dynamics of the invasive Asian tiger mosquito Aedes albopictus are strongly affected by winter. The work we present here analyzes the adaptive synchronization of the diapause process in the wintry generation of A. albopictus, where the egg stage is exposed to adverse winter conditions. The seasonal pattern of egg laying activity of a French Mediterranean population of the Asian tiger mosquito was monitored weekly for 2 years with ovitraps. The field diapause incidence and the critical photoperiod (CPP, i.e. the maternal day length inducing diapause in 50% of the eggs), were determined by hatching experiments on the collected eggs. The period of diapause termination was estimated by a field survey of the first hatchings for both years. The CPP is equal to 13.5 hours of light and occurs in the field on the 25th of August. Thus, it is on September 11th, 17 days after the CPP, that 50% of the eggs are in a prediapause stage in the field. The egg diapause rate increases rapidly during September, whereas the mean number of eggs laid decreases sharply after mid-September. Surprisingly, after having reached a peak of 95% at the end of September, from mid-October the diapause incidence declined and stayed below 50%. Indeed, both years the diapause initiates before the rapid decrease of the environmental temperature. This leaves a sufficient period of time to the complete development of one generation of A. albopictus with effective induction of diapause in the laid eggs. The very first larvae hatched were sampled both years in the first half of March. With 20 to 26 weeks in the egg stage and about 7 weeks in the larval stages, the first annual generation spends a long time in immature stages. On a practical point of view, this long development time represents a wide window for eggs and larvae control in early spring.

When mothers anticipate: Effects of the prediapause stage on embryo development time and of maternal photoperiod on eggs of a temperate and a tropical strains of Aedes albopictus (Diptera: Culicidae)

BackgroundThe diapause of Aedes albopictus is maternally induced by photoperiod and initiates at the pharate larvae stage in eggs. This pre-diapause results in enhanced survival eggs during the winter. This study aims to disentangle the effects of photoperiod and diapause on embryonic developmental time and egg size in A. albopictus. A temperate strain capable to perform diapause and a tropical strain unable of diapause were reared at 21°C with long-(LD) and short-day (SD) lengths. Four distinct traits were studied on embryos and eggs were measured at the end of embryogenesis. ResultsThe chronologies of embryo development for both strains were influenced by maternal photoperiod, especially in the temperate strain in which the development of SD eggs took longer than LD eggs. The delay increased gradually in the temperate strain, and reached up to 38h at the end of embryogenesis. The kinetics of embryogenesis differed among the temperate and the tropical strains, each one of the 4 studied traits showing differences. For example the serosal cuticle was secreted precociously in the tropical strain. Egg width and volume are influenced by the maternal photoperiod and the strain×photoperiod interaction. For both strains, larger eggs were laid by female reared under SD when compared to LD. ConclusionsThe influence of several maternal effects was demonstrated in this study. The diapause process modifies greatly the length of embryogenesis in the temperate strain, whereas the maternal photoperiod has a direct influence on egg size and embryogenesis regardless of the strain considered. These findings provide useful data on chronology of embryonic development for integrative biology studies of egg pre-diapause stages.

The differences in metabolic activity of subitaneous and diapause eggs in Centropages tenuiremis

The respiration and the isoenzymes of subitaneous and diapause eggs of Centropages tenuiremis in Xiamen waters of China were investigated. The results demonstrated that the average oxygen consumption of subitaneous eggs was lower than that of diapause eggs during the pre-diapause stage. The character of respiration in diapause eggs was as follows: the oxygen consumption quickly rose after spawning, and reached a maximal value, then gradually declined to a lower value that was maintained at a lower value (� 20% of the maximal value), the low value of oxygen consumption in diapause eggs was lower than that in subitaneous eggs at the same temperature (� 70%). The experimental results also indicated that among subitaneous, diapause and pre-diapause eggs of Centropages tenuiremis, the pre-diapause eggs were the most sensitive to temperature, the diapause eggs were the most insensitive; the sensitivity of subitaneous eggs to temperature was intermediate. The results also showed that there are obvious differences of isozyme patterns in malate dehydrogenase (MDH), sorbitol dehydrogenase (SDH), superoxide dismutase (SOD) and esterase (EST) between subitaneous eggs and pre-diapause eggs, and the characteristic enzyme bands for pre-diapause eggs were shown to be as follows: MDH 61 ,M DH 81 , SDH 41 , SDH 60 , SOD 97 , EST 48 , EST 59 and EST 61 ; for subitaneous eggs: MDH 59 , SDH 70 and EST 93 .

Optimal diapause strategies of a grasshopper, Melanoplus sanguinipes

Previous analyses of diapause in insects have most often focused on the timing of the switch from non-diapausing to diapausing offspring in bivoltine populations and have assumed that diapause is irreversible or that the insect cannot survive winter if not in diapause. Many insects exhibit more flexibility in their life cycles, such as the age at which diapause begins, and facultative diapause, that may influence the evolution of different diapause strategies in different environments. The grasshopper Melanoplus sanguinipes F. (Orthoptera: Acrididae), has a very wide geographic range over which diapause characteristics vary greatly. Embryonic diapause in this species may be under maternal control, may be obligate or facultative (i.e., may be averted by cold temperature treatment of pre-diapause embryos), and embryos may enter diapause at different ages. Diapause traits were examined in two populations of M. sanguinipes from very different environments. In the population from a temperate climate (Idaho, USA), diapause was facultative, i.e., pre-diapause embryos averted diapause when held at 5° C for 90 days at all ages tested (7 days and older). The Idaho embryos entered diapause in late stage of development if held at 22° C for 30 days or more. In populations from subarctic Alaska, USA, embryos also entered diapause in a late stage of development, but diapause was obligate and could not be averted by chilling in the pre-diapause stages. Simulated evolution of these traits over a wide range of season-lengths showed that late stage diapause is an essential trait in very short season environments, resulting in early hatching, and a semivoltine life-cycle. Facultative diapause enabled bivoltinism to be a viable strategy in shorter seasons than when diapause was obligate. At transitions from semivoltine to univoltine, and from univoltine to bivoltine life cycles, populations with obligate diapause adopted a strategy of no diapause (via maternal effects) to enable univoltine life cycles.

Effect of anoxia on diapause termination in eggs of the false melon beetle, Atrachya menetriesi

The effect of anoxia on diapause development in the leaf beetle Atrachya menetriesi was investigated to elucidate the role of oxygen in regulation of egg diapause. While anoxia alone had no effect on diapause termination, it decreased diapause intensity before chilling. Such an effect reached a maximum level when anoxia lasted for about 10 days. Anoxia applied during the pre-diapause stage also reduced diapause intensity. On the other hand, anoxia terminated diapause when the diapause intensity had been lowered by sufficient duration of chilling (50 days at 7.5 °C). The effect of anoxia was temperature dependent; the larger effect was elicited when anoxia was combined with a higher temperature. A 50-day chilling caused more than 20% of eggs to terminate diapause upon transfer to warm conditions. However, when this chilling period was interrupted on the 20th day by a 5-day exposure to a high temperature of 20, 25 or 30 °C, the effect of the former chilling was cancelled partially or completely, suggesting that warming reversed diapause development. This reversing effect of a high temperature, however, was not manifested when the warming was combined with anoxia. The results suggest that anoxia inhibits diapause reversal and facilitates a certain process of diapause development. The sequence of exposure to anoxia and chilling is not important.

Variation in diapause potential and strength in eggs of the Australian plague locust, Chortoicetes terminifera (Walker) (Orthoptera: Acrididae)

Samples of eggs of Chortoicetes terminifera were incubated under 3 temperature regimes which would allow non-diapause eggs to develop about 50% and so take them beyond the diapause stage. Even so, many more eggs entered diapause when reared at 20°C for 3 weeks than at 32°C for 1 week. By collecting and incubating eggs at intervals after laying in autumn, diapause potential or strength of eggs at different stages of development was estimated. For eggs in pre-diapause, the proportion in diapause after rearing was used to estimate diapause “potential”; for those in diapause, the proportion was used to estimate diapause “strength”. At laying, eggs varied greatly in their potential to enter diapause. However, those with lower potential at laying often increased their potential during the pre-diapause stage so that by the time diapause was reached diapause strength varied but over a lesser range. In all eggbeds, diapause strength decreased by 7–9 weeks after laying; little or none remained by mid-winter. These variations in diapause potential and diapause strength seem to reflect how much the temperature threshold for development during diapause is increased above that for non-diapause. Both diapause potential and strength may reflect the value of some factor whose level at laying is determined by the environment experienced by the parents but which changes subsequently.

Egg Diapause and Water Absorption in the False Melon Beetle, Atrachya menetriesi FALDERMANN(Coleoptera : Chrysomelidae)

The relation between the egg diapause and water absorption was investigated in Atrachya menetriesi. The egg absorbed water in the post-diapause stage but not in the pre-diapause stage. When the non-diapuase and diapause-terminated eggs were kept wet, they absorbed water at a constant rate and their weight increased by about 80% just before hatching. If the diapausing egg was kept wet, it absorbed water very slowly, but the embryo remained in the definite diapause stage and further develpment was not discernible. On the other hand, if the non-diapause egg was not in contact with water from the time of oviposition, the embryo ceased to develop at a stage more advanced than the diapause stage and survived for a certain period. If water was not accessible to the diapause-terminated egg, the embryo was able to resume growth but did no develop beyond the spiracle-formation stage.It seems therefore unlikely that the egg diapause is caused by water deficiency.

The Action of Urea on Diapause in Eggs of Acheta Commodus (Walk.) (Orthoptera: Gryllidae)

U rea, at concentrations of 0�04-0, 08M, was found to prevent the onset of diapause in eggs of Acheta commodu8 (Walk.) when applied during the prediapause stages of development.

INVESTIGATIONS OF PERMEABILITY, DIAPAUSE, AND HATCHING IN THE EGGS OF THE MOSQUITO AEDES HEXODONTUS DYAR

Methods for collection in the laboratory of many thousands of eggs from field-collected, blood-fed females are described. Also a way of separating fertile from infertile eggs is outlined. When the eggs are laid they are white but begin to darken within one-half hour. The darkening did not occur when the egg cell or early embryo was killed.Two changes in permeability in the prediapause stage of the eggs of Aedcs hexodontus were observed. The first took place as the chorion changed from white to black. The egg lost and gained water easily when first laid but resisted water loss and uptake as the chorion darkened. However, a complete waterproofing did not result. With the formation of the transparent cuticle a further change in permeability occurred. The egg lost hardly any water even when exposed to extreme desiccation and what little water was lost was regained very slowly. However, the cuticle, with the chorion removed in sodium hypochlorite, was quite permeable. The impermeability of the chorion and cuticle combination must result from the bonding between the two and this bonding must be disrupted by the dechorionation.To study the obligate diapause that occurs in the late embryogeny of the mosquito egg the effect of making water available to the embryo, of light, and of low temperature was studied. Diapause was not broken when water was available to the embryo, nor did alternate periods of light and dark at room temperature or at lower temperatures disrupt the diapause. To test the influence of cold the temperature of 1° and −3 °C. were used to simulate the temperatures in the field. The low temperatures were found to terminate the diapause but the numbers of eggs hatching did not steadily increase with an increase in the time of exposure to cold. No clue has as yet been discovered to explain these results. A hatching infusion of decaying adult mosquito bodies in distilled water was found to give more hatching than did distilled water alone. The mechanism of the hatching stimulus is not known.