Ontogenetic patterns of cold-hardiness and glycerol production in Sarcophaga crassipalpis

Developmental patterns of low-temperature tolerance and glycerol production were determined for larval, pupal and adult stages of the flesh fly Sarcophaga crassipalpis Macquart (Diptera: Sarcophagidae). Both diapause and non-diapause-destined flies were reared at relatively high temperatures, 20° or 25°C, prior to testing. Cold tolerance was greatest for diapause pupae aged 12–35 days after pupariation. Among non-diapause-destined flies, pupae exhibited a greater level of low temperature tolerance than larvae or adults. Although diapause pupae were more tolerant than non-diapause pupae maximal cold tolerance was not attained in either group until 10 days after pupariation. Non-diapause-destined feeding and wandering larvae had higher glycerol levels than larvae destined for diapause. During the first 6 weeks after pupariation glycerol titres increased steadily in diapause pupae. Rapid loss of glycerol is associated with the termination of pupal diapause.     

Comparison of the circadian eclosion rhythm between non-diapause and diapause pupae in the onion fly, Delia antiqua: the change of rhythmicity

When pupae of Delia antiqua were transferred to constant darkness (DD) from light-dark (LD) cycles or constant light (LL), the sensitivity to light of the circadian clock controlling eclosion increased with age. The daily rhythm of eclosion appeared in both non-diapause and diapause pupae only when this transfer was made during late pharate adult development. When transferred from LL to DD in the early pupal stage, the adult eclosion was weakly rhythmic in non-diapause pupae but arrhythmic in diapause pupae. However, the sensitivity of the circadian clock to temperature cycles or steps was higher in diapause pupae than in non-diapause pupae; in the transfer to a constant 20 degrees C from a thermoperiod of 25 degrees C (12 h)/20 degrees C (12 h) on day 10 after pupation or from chilling (7.5 degrees C) in DD, the adult eclosion from diapause pupae was rhythmic but that from non-diapause pupae arrhythmic. In a transfer to 20 degrees C from the thermoperiod after the initiation of eclosion, rhythmicity was observed in both types of pupae. The larval stage was insensitive to the effect of LD cycle initiating the eclosion rhythm. In D. antiqua pupae in the soil under natural conditions, therefore, the thermoperiod in the late pupal stage would be the most important 'Zeitgeber' for the determination of eclosion timing.     

Diapause development in Bombyx eggs in relation to ‘esterase A’ activity

Diapause is broken by hydrochloric acid treatment and also terminated by long chilling of eggs in the silkworm, Bombyx mori. One of esterases in silkworm eggs named ‘esterase A’ is closely related to diapause of this insect.Hydrochloric acid treatment of diapause eggs induced a prompt elevation of esterase A activity. The elevation was observed within 30 min after treatment. This HCl treatment effectively stimulated the eggs to hatch. This indicates that the increase of esterase A activity is correlated with an active resumption of morphogenesis.The question was examined of whether chilling also increases esterase A activity or not. It was found that chilling also caused an increase of esterase A activity. This increase occurred before the re-appearance of glycogen in eggs, which indicates the termination of diapause in this insect. In fact, the establishment of hatchability in chilled eggs was observed after esterase A activity has reached the maximum level. Thus the increase of esterase A activity could be regarded as associated with the termination of diapause per se but not with the subsequent process of post-diapause development. This change in esterase A activity was observed only in chilled diapause eggs and was not observed in diapause eggs without chilling and non-diapause eggs.These results suggest that the increase of esterase A activity during chilling may be a kind of activity that occurs during the diapause stage in preparation for resumption of morphogenesis or diapause development.     

Haemocytes of the pink bollworm, Pectinophora gossypiella, during larval development and diapause

Seven types of haemocytes were observed in the last larval instar of the pink bollworm, Pectinophora gossypiella (Saunders): prohaemocytes, plasmatocytes, granular haemocytes, spherule cells, adipohaemocytes, oenocytoids, and podocytes. Total and differential haemocyte counts made from diapausing and non-diapausing larvae showed that during diapause there was a significant reduction in the numbers of all haemocyte types. Upon termination of diapause, the haemocyte level increased. There were no significant differences in the level of haemocytes in the pharate pupae that developed from diapause or non-diapause type larvae, except in the case of adipohaemocytes, which were three times as prevalent in pharate pupae from diapausing larvae. Functional aspects of various types of haemocytes are discussed, and it is suggested that the lower haemocyte level observed during diapause is the result of lower metabolic activity.     

Light and electron microscopic studies on the neurosecretory control of diapause incidence in Pieris rapae crucivora

The incidence of diapause was shown to be determined humorally during the larval-pupal ecdysis by means of brain extirpation experiments.On the basis of this observation, light and electron microscopic changes in the neurosecretory type II cells in the pars intercerebralis-corpus cardiacum system during pharate pupal and early pupal stages were examined in insects reared under long day-length (non-diapause individuals) and in insects reared under short day-length (diapause individuals). In the diapause individuals, neurosecretory granules in NS-II cells increased during the pupal instar and large aggregates of granules packed the cytoplasm. Thereafter, inclusion bodies showing cytoplasmic breakdown of the granules appeared.In the non-diapause individuals, on the contrary, electron micrographs suggesting the release of neurosecretory material from axon terminals were obtained just after the pupal ecdysis. There were very few granules, with many Golgi bodies and much rough ER 8 to 12 hr after the ecdysis.It is concluded that adult development is determined by the release of neurosecretory material from the axon terminals of NS-II cells at the larval-pupal ecdysis. If release does not occur, the pupae enter diapause. It is also thought that differences in day-length during the larval stages influence the activities of NS-II cells before pupation.     

Endocrine Mechanisms Regulating Post-Diapause Development in the Cabbage Armyworm,Mamestra brassicae

Diapause, a programmed developmental arrest at a specific stage, is common in insects and is regulated by hormones. It is well established that in pupal diapause, cessation of ecdysteroid secretion from the prothoracic glands (PGs) after pupal ecdysis leads to diapause initiation, while resumption of its secretion induces post-diapause development. However, what regulates the activity of the glands is poorly understood, especially for the glands of diapause-terminated pupae. In the present study, we investigate the mechanisms by which post-diapause development is regulated in the cabbage armyworm Mamestra brassicae. We demonstrate that the brain is necessary for the initiation of post-diapause development and that the factor in the brain responsible for the activation of the PGs is the prothoracicotropic hormone (PTTH). Further, through measuring the hemolymph PTTH titers by time-resolved fluoroimmunoassay, we show that PTTH is actually released into the hemolymph prior to the activation of the PGs. Although its peak titer is much lower than expected, this low concentration of PTTH is most likely still effective to activate the PGs of post-diapause pupae, because the responsiveness to PTTH of the glands at this stage is very high compared to that of nondiapause pupal PGs. These results strongly suggest that in M. brassicae, PTTH serves as a trigger to initiate pupa-adult development after diapause termination by stimulating the PGs to secrete ecdysteroid.     

Effect of photoperiod on the induction and maintenance of diapause and on development of the predatory bugPodisus maculiventris (Hem.: Pentatomidae)

The predatory bugPodisus maculiventris Say displays a reproductive diapause. In the laboratory, at 23±1°C, when the pre-imaginal stages were reared on larvae ofGalleria melonella L. (Lepidoptera: Pyralidae) under a short photoperiod of 8L: 16D, diapause was induced in all adult females, whereas under a long photoperiod of 16L: 8D none entered diapause. Nymphal development was faster under the diapause-inducing short photoperiod than under the diapause-averting long photo-period. By contrast, embryonic development was faster under the long than under the short photoperiod. Diapause maintenance was also under the control of photoperiod. After a 10-d-period of chilling at 4°C, diapause was quickly terminated in adults kept under long photoperiod whereas it was maintained in most of those kept under short photoperiod. The predation rate of non-diapause or post-diapause adult females was much higher than that of diapause females.     

Maternal influence on the incidence and duration of larval diapause in Calliphora vicina

ABSTRACT. The photoperiodic mechanism governing larval diapause in the blowfly Calliphora vicina R.-D. (Diptera: Calliphoridac) is largely maternal. The critical daylength for adult flies at 20C, and larvae derived from day 12 eggs and maintained in darkness at 11C, is about 14.5 h/24. At short daylength (LD 12:12 h) and a range of temperatures, eggs laid on or before day 9 gave rise to a low incidence of diapause, whereas those laid on day 10 and later gave rise to a high incidence; the photoperiodic 'counter' therefore shows a temperature-compensated mechanism for 'accumulating' successive short days. The rate of switching from non-diapause to diapause progeny is hardly affected by the timing of protein meals, although an early supply of meat accelerates egg development and leads to an increasing proportion of early, non-diapausing progeny. There is no evidence for a maternal effect preventing diapause induction in the first post-diapause ('spring') generation of flies. Diapausing larvae terminate diapause spontaneously in darkness at 11C after a variable period which is positively correlated with the incidence of diapause in the group. Diapause incidence and duration therefore are both determined maternally.     

Comparison of the circadian eclosion rhythm between non-diapause and diapause pupae in the onion fly,Delia antiqua

The influence of pupal diapause on adult eclosion rhythm of Delia antiqua was investigated. When non-diapause and diapause pupae were exposed to various photoperiods at 15, 20 and 25 °C, both of them emerged as adults close to the light-on time, but the phase of eclosion varied with photoperiod and temperature. Moreover, there was a significant difference in the eclosion time between non-diapause and diapause pupae; the eclosion peak of diapause pupae was earlier than that of non-diapause pupae. When non-diapause and diapause pupae were transferred to constant darkness (DD) after having experienced LD 12:12 at 15, 20 and 25 °C, both showed circadian rhythmicity in eclosion. Although the free-running period (τ) decreased slightly as temperature increased in both non-diapause and diapause pupae, the latter tended to show shorter τ than the former. This observation suggests that the observed difference in eclosion time in LD cycles between non-diapause and diapause pupae is due to differences in τ.     

草地螟滞育幼虫的蛋白和核酸含量变化

为了阐明草地螟Loxostege sticticalis 滞育的分子调控基础, 本研究应用Trizol法、 DNA和蛋白质定量试剂盒、 SDS-PAGE电泳技术分别对进入滞育1, 2, 3和4个月、 解除滞育以及非滞育草地螟老熟幼虫中的核酸含量、 总蛋白含量和组分的变化进行了测定。结果表明： 滞育不同月份幼虫的总RNA含量显著低于非滞育幼虫的总RNA含量（P＜0.05）; 解除滞育幼虫的总RNA含量显著高于滞育2, 3和4个月的幼虫, 但仍显著低于非滞育的对照组。滞育不同月份、 非滞育以及解除滞育幼虫的总DNA含量没有显著差异（P＞0.05）。RNA/DNA比值随滞育的开始而显著下降, 随着滞育的结束而显著上升。滞育不同月份的幼虫总蛋白含量显著高于非滞育幼虫的总蛋白含量（P<0.05）, 但解除滞育与非滞育幼虫的总蛋白含量无显著差异。利用SDS-PAGE电泳分析发现滞育幼虫体内存在滞育关联蛋白, 蛋白条带在24 kDa左右, 但非滞育和已经解除滞育的幼虫则没有该蛋白条带。这些结果表明, 总RNA含量的降低、 RNA/DNA比值下降、 总蛋白含量的升高, 以及24 kDa蛋白的存在是草地螟幼虫滞育的主要生理特征。     

Control of summer and winter diapause in the leaf-mining fly Pegomyia bicolor Wiedemann (Dipt., Anthomyiidae)

Effects of photoperiod and temperature on diapause induction and termination were investigated in both aestival and hibernal pupae of Pegomyia bicolor Wiedemann under field and laboratory conditions. In the field, summer diapause had occurred already in part of the first pupal population; the proportion of diapause gradually rose as the day length and temperature increased. This fly is a short-day species with a pupal summer and winter diapause. Summer diapause was induced by both long day-lengths and mild temperatures. The whole larval life is sensitive to photoperiod. Winter diapause was induced mainly by low temperatures, especially in the first 10 days after pupation. High temperatures strongly enhanced summer diapause induction regardless of photoperiod. The diapause-averting influence of short photoperiods was fully expressed only at moderately low temperatures. High temperatures delayed diapause development, resulting in a rather long summer diapause; whereas low temperatures hastened it, leading to a short winter diapause and showing a low thermal threshold for diapause development. In the field, the post-diapause development started in January, the coldest month, suggesting that the thermal requirements for post-diapause development is also low.     

Hormonal control of seasonal morph determination in the swallowtail butterfly, Papilio xuthus L. (Lepidoptera: Papilionidae)

Seasonal morphs (spring and sumer forms) of Papilio xuthus L. are determined coincidentally with diapause and non-diapause in pupae by larval exposure to short days and long days respectively. The neuroendocrine principle underlying seasonal-morph determination was studied using surgical operations in P. xuthus.When recipient 0-day old or chilled diapause pupae were joined to donor 0-day old non-diapause pupae, the recipients developed into summer or intermediate morphs. When the same kinds of recipients used above were joined to 0-day old or chilled diapause pupae, there were no significant effects on the adult morph. In contrast, recipient non-diapause pupae all developed into summer morphs, regardless of groups of the type of donors.Furthermore, when the brain of 5th-instar larvae, pharate pupae or pupae, predetermined to be diapause, was transplanted into the abdomen of 0-day old, 30-day old or chilled diapause pupae, the recipients developed into summer or intermediate morphs.The results indicate that the brain of non-diapause pupae secretes a humoral factor producing the summer morph. In non-diapause pupae, the factor may be secreted at about the stage of larval-pupal ecdysis coincidentally with that of prothoracicotropic hormone.     

Photoperiodic sensitivity and diapause induction during ovarian,embryonic and larval development of the flesh fly,Sarcophaga argyrostoma

Sensitivity to the daily photoperiod, particularly with respect to pupal diapause induction, was studied during ovarian, embryonic, and larval development of the flesh flySarcophaga argyrostoma. Large flies were shown to have a greater number of primary follicles in their ovaries and to be capable of limited ovarian maturation in the absence of exogenous protein (autogeny). Such ovarian development occurred independently of photoperiod. However, long days experienced during embryogenesis caused more rapid development, and earlier larviposition, than short days. Short days during embryonic and subsequent larval development also induced pupal diapause, whereas long days led to continuous or non-diapause development of the pupae. Pupal diapause could not be induced by photoperiods during the vitellogenic phase of ovarian development. InSarcophaga argyrostoma, a maternal effect preventing pupal diapause among the progeny of files with a diapause history was not observed.     

Control of diapause and supercooling by the retrocerebral complex inPyrrhocoris apterus

Effects of external (photoperiod, temperature) and internal (retrocerebral complex, diapause condition) factors on supercooling were studied inPyrrhocoris apterus (L.) (Heteroptera: Pyrrhocoridae) adults. An increase in supercooling capacity is associated with the induction of diapause by short-day photoperiod or cardiacallatectomy of long-day non-diapause insects in spite of a high temperature of 26°C. The induction of diapause is a prerequisite for a further increase in supercooling capacity by cold acclimation. Post-diapause adults show low values of the supercooling point also in late January or early February, although their developmental potential is fully restored. Evidently the ability to supercool associated with diapause induction is maintained by a low ambient temperature in spite of diapause termination. The supercooling point increases rapidly when these adults are transferred to a high temperature of 26°C and cold re-acclimation is then no longer possible. The inhibition of morphogenesis (maturation of gonads) by cardiacallatectomy does not prevent the supercooling point from increasing in post-diapause adults at the high temperature.     

Role of the brain in post-diapause adult development in the swallowtail, Papilio xuthus

The role of the brain in adult development was examined by brain removal in unchilled and chilled diapausing pupae of Papilio xuthus. Chilling was effective in shortening the pupal duration and synchronizing adult emergence, although photoperiod had little effect on diapause development. The brain played a role of shortening the pupal duration and synchronizing adult emergence in both unchilled and chilled individuals, although it was not essential for post-diapause adult development. The stimulus of low temperature was recorded even in the absence of the brain, because chilling shortened the pupal duration in brainless individuals. The brain showed activity which affected subsequent adult development in chilled pupae within one day after chilling in males. This period was less limited in females.     

Diapause development in the chestnut weevilCurculio elephas

Larval diapause development in the chestnut weevilCurculio elephas (Coleoptera, Curculionidae) was studied in the laboratory at different temperatures. The results proved that exposure to low temperatures (3–6°C) in the period December–February is not required to complete diapause. The diapause is terminated in December and from January on, the larvae can initiate post-diapause morphogenesis in the laboratory, if temperatures allow it. In the field developmental rates are negligible during winter cold (4–6°C) and only after March morphogenesis can proceed with no interruption until adult emergence. Diapause and post-diapause quiescence contribute to individual synchronization for initiation of development. The observed spread of adult emergence was 30 days in the laboratory. This variability produced during post-diapause development may be a response to annual variation in the phenology of the chestnuts.     

Comparison of the circadian eclosion rhythm between non-diapause and diapause pupae in the onion fly,Delia antiqua: the effect of thermoperiod

When non-diapause and diapause pupae of Deliaantiqua were exposed to various thermoperiods where thermophase (T) was 25 °C and the cryophase (C) was 15 or 20 °C (TC₁₅ or TC₂₀) in constant darkness (DD), the majority of both types of flies emerged before the rise in temperature. Eclosion time was delayed at the lower cryophase temperature. Moreover, there was a significant difference in the time of adult eclosion between non-diapause and diapause pupae; diapause pupae eclosed earlier than non-diapause pupae. When the two types of pupae were transferred to a constant low temperature (15 or 20 °C) after having experienced TC₁₅ or TC₂₀ 12:12 h, they showed circadian rhythmicity in eclosion. The free-running period (τ) of the eclosion rhythm changed after transfer to constant low temperatures in both non-diapause and diapause pupae, suggesting that this change represents a transient cycle until the temperature-sensitive oscillator is coupled again to the temperature-insensitive pacemaker. However, diapause pupae tended to show a shorter τ than non-diapause pupae. This observation suggests that the difference in adult eclosion time under thermoperiodic conditions between non-diapause and diapause pupae is related to their different τ s.     

Host plant and latitude-related diapause variation in Rhagoletis pomonella: a test for multifaceted life history adaptation on different stages of diapause development

Variation in the overwintering pupal diapause of Rhagoletis pomonella appears to adapt sympatric populations of the fly to seasonal differences in the fruiting times of their host plants, generating ecological reproductive isolation. Here, we investigate what aspects of diapause development are differentially affected (1) by comparing the propensities of apple vs. hawthorn-infesting host races of R. pomonella to forgo an initially deep diapause and directly develop into adults, and (2) by determining the chronological order that R. pomonella races and sibling species break diapause and eclose when reared under standardized environmental conditions. The results imply that factors affecting initial diapause depth (and/or differential mortality during the prewintering period) and those determining the timing of diapause termination or rates of post-diapause development are both under differential selection and are to some degree genetically uncoupled in flies. The modular nature of diapause life history adaptation in Rhagoletis suggests that phenology may involve multiple genetic changes and represent a stronger ecological barrier separating phytophagous specialists than is generally appreciated.