Diapause Intensity Research Articles

Crowd-sourced observations of a polyphagous moth reveal evidence of allochronic speciation varying along a latitudinal gradient.

Allochronic speciation, where reproductive isolation between populations of a species is facilitated by a difference in reproductive timing, depends on abiotic factors such as seasonality and biotic factors such as diapause intensity. These factors are strongly influenced by latitudinal trends in climate, so we hypothesized that there is a relationship between latitude and divergence among populations separated by life history timing. Hyphantria cunea (the fall webworm), a lepidopteran defoliator with red and black colour morphs, is hypothesized to be experiencing an incipient allochronic speciation. However, given their broad geographic range, the strength of allochronic speciation may vary across latitude. We annotated >11,000 crowd-sourced observations of fall webworm to model geographic distribution, phenology, and differences in colour phenotype between morphs across North America. We found that red and black morph life history timing differs across North America, and the phenology of morphs diverges more in warmer climates at lower latitudes. We also found some evidence that the colour phenotype of morphs also diverges at lower latitudes, suggesting reduced gene flow between colour morphs. Our results demonstrate that seasonality in lower latitudes may increase the strength of allochronic speciation in insects, and that the strength of sympatric speciation can vary along a latitudinal gradient. This has implications for our understanding of broad-scale speciation events and trends in global biodiversity.

Cold tolerance and diapause within and across trophic levels: Endoparasitic wasps and their fly host have similar phenotypes

Low temperatures associated with winter can limit the survival of organisms, especially ectotherms whose body temperature is similar to their environment. However, there is a gap in understanding how overwintering may vary among groups of species that interact closely, such as multiple parasitoid species that attack the same host insect. Here, we investigate cold tolerance and diapause phenotypes in three endoparasitoid wasps of the apple maggot fly Rhagoletis pomonella (Diptera: Tephritidae): Utetes canaliculatus, Diachasma alloeum, and Diachasmimorpha mellea (Hymenoptera: Braconidae). Using a combination of respirometry and eclosion tracking, we found that all three wasp species exhibited the same three diapause duration phenotypes as the fly host. Weak (short duration) diapause was rare, with <5 % of all three wasp species prematurely terminating diapause at 21 °C. Most D.mellea (93 %) entered a more intense (longer duration) diapause that did not terminate within 100 d at this warm temperature. The majority of U.canaliculatus (92 %) and D. alloeum (72 %) averted diapause (non-diapause) at 21 °C. There was limited interspecific variation in acute cold tolerance among the three wasp species: wasps and flies had similarly high survival (>87 %) following exposure to extreme low temperatures (−20 °C) as long as their body fluids did not freeze. The three wasp species also displayed little interspecific variation in survival following prolonged exposure to mild chilling of 8 or more weeks at 4 °C. Our study thus documents a remarkable conservation of cold tolerance and diapause phenotypes within and across trophic levels.

Seasonal and geographical adaptations in the parthenogenetic stick insect, Ramulus mikado (Phasmatodea: Phasmatidae)

Seasonal and geographical adaptations in terms of obligatory embryonic diapause in the parthenogenetic stick insect, Ramulus mikado, were studied. First and second instar nymphs were collected at locations at three latitudes in Japan and reared in the laboratory under a photoperiod of 16L : 8D or 12L : 12D at 25°C. Their eggs were kept at 30°C for 30 or 60 days after oviposition, but no eggs hatched. Hatching was observed more than 100 days after transfer from 30°C to 15°C. The long period between transfer and hatching indicate that eggs in an early embryonic stage of development enter diapause at high-temperatures. The time from oviposition to hatching of eggs laid by adults that originated from the three locations kept under constant conditions between 15 and 25°C were compared. In all these experiments, eggs laid by individuals originating from high latitudes took longer to hatch. The eggs of those originating from Okayama and Ehime did not hatch at 25°C. However, more than 80% of the eggs of those that originated from the northernmost population hatched. Hatching before winter was observed when the eggs of those that originated from the northern population were placed outdoors in Okayama, even when the maternal insects were reared under long-day conditions in the laboratory. These findings indicate that univoltine R. mikado enters diapause twice during embryonic development, which enables it survive adverse conditions in summer and winter, respectively. Furthermore, diapause intensity was lowest in insects that originated from the lowest latitude.

Genomically correlated trait combinations and antagonistic selection contributing to counterintuitive genetic patterns of adaptive diapause divergence in Rhagoletis flies.

Adaptation to novel environments can result in unanticipated genomic responses to selection. Here, we illustrate how multifarious, correlational selection helps explain a counterintuitive pattern of genetic divergence between the recently derived apple- and ancestral hawthorn-infesting host races of Rhagoletis pomonella (Diptera: Tephritidae). The apple host race terminates diapause and emerges as adults earlier in the season than the hawthorn host race, to coincide with the earlier fruiting phenology of their apple hosts. However, alleles at many loci associated with later emergence paradoxically occur at higher frequencies in sympatric populations of the apple compared to the hawthorn race. We present genomic evidence that historical selection over geographically varying environmental gradients across North America generated genetic correlations between two life history traits, diapause intensity and diapause termination, in the hawthorn host race. Moreover, the loci associated with these life history traits are concentrated in genomic regions in high linkage disequilibrium (LD). These genetic correlations are antagonistic to contemporary selection on local apple host race populations that favours increased initial diapause depth and earlier, not later, diapause termination. Thus, the paradox of apple flies appears due, in part, to pleiotropy or linkage of alleles associated with later adult emergence and increased initial diapause intensity, the latter trait strongly selected for by the earlier phenology of apples. Our results demonstrate how understanding of multivariate trait combinations and the correlative nature of selective forces acting on them can improve predictions concerning adaptive evolution and help explain seemingly counterintuitive patterns of genetic diversity in nature.

Plasticity of nutrient accumulation patterns in diapausing fall webworm pupae.

The accumulation of nutrients during diapause preparation is crucial because any lack of nutrition will reduce the likelihood of insects completing diapause, thereby decreasing their chances of survival and reproduction. The fall webworm, Hyphantria cunea, diapause as overwintering pupae and their diapause incidence and diapause intensity are regulated by the photoperiod. In this study, we test the hypothesis that photoperiod influences energy reserve accumulation during diapause preparation in fall webworm. We found that the body size and mass, lipid and carbohydrate content of pupae with a short photoperiod during the diapause induction phase were significantly greater than those of pupae with a relatively short photoperiod, and the efficiency of converting digested food and ingested food into body matter was greater in the short-photoperiod diapause-destined larvae than the relatively short-photoperiod diapause-destined larvae. We also observed higher lipase and amylase activities in short-photoperiod diapause-destined larvae relative to the counterparts. However, no obvious difference was found in protein and protease in the pupae with a short photoperiod during the diapause induction phase and short-photoperiod diapause-destined larvae compared with the counterparts. Therefore, we conclude that the energy reserve patterns of diapausing fall webworm pupae are plastic and that short-photoperiod diapause-destined larvae increase their energy reserves by improving their feeding efficiency and increase their lipid and carbohydrate stores by increasing the lipase and amylase activities in the midgut.

Effect of photoperiod on winter and summer diapause of the soybean pod borer Leguminivora glycinivorella (Lepidoptera: Tortricidae)

• Soybean pod borer larvae continuously undergo winter and summer diapause. • Winter diapause intensity is weakened by January without photoperiodic responses. • Summer diapause is maintained by long day length. • Summer diapause ends with photoperiodic transition from long to short day length. • Males and females experiencing photoperiodic transition emerge synchronously. To understand the geographical differences between diapause systems and synchronization of adult occurrence in the soybean pod borer Leguminivora glycinivorella (Lepidoptera: Tortricidae), we examined the timing of winter diapause termination and intensity of summer diapause using univoltine and potentially bivoltine individuals in Iwate, Japan. In laboratory rearing experiments of mature larvae maintained at constant temperature (20 °C), winter diapause intensity weakened by January without photoperiodic responses. Meanwhile, summer diapause was maintained by the long day length and presumably terminated with the photoperiodic transition from long to short day length. The intensity of summer diapause was stronger for cocoons that transitioned from a 16 h light to 8 h dark (LD 16:8) to a LD 15:9 photoperiod than for those that transitioned from LD 15:9 to LD 14:10. These results suggest that populations distributed in relatively low-latitude areas, with partly or potentially bivoltine individuals, would have a weaker summer diapause or none at all. Moreover, sexual differences in the number of days to emergence were not detected when individuals experienced a photoperiodic transition from long to short day length, suggesting that the summer diapause system may function to synchronize the emergence of males and females in the population examined.

English

The solitary endoparasitoid, Microplitis rufiventris is considered one of the principal wasps on the cotton leaf moth, Spodoptera littoralis larvae. It has a strange sort of obligate diapause. The data present an evidence of persistent low-level diapause throughout the year and every year that observed under mass rearing conditions. When continuous rearing was done throughout the year at 25&deg;C and 12/12 h L/D, a small portion of the individuals in each large samples of each generation entered diapause. So, a long-timed experiment that repeated three years and collecting dataset that required regular extensive dissections and microscopic observations of emerged diapausing wasps was carried out. This diapause spontaneously terminates after time periods similar each year, showing an evidence of &ldquo;time memory&rdquo; in diapausing wasp behavior. The duration of this time period is taken as a proxy of diapause intensity. This had occurred whether the large number of individuals originated in summer from field-collected host larvae or from three successive years of laboratory-rearing parasitoids. Contrary to non diapausing wasps, diapausing females emerge some days early than males. This is the first report of new sort of obligate diapause in diapause induction and termination in M. rufiventris under the same constant rearing conditions. Fecundity of diapaused wasps was diapause intensity dependent. Wasps diapaused for 60-86 days (long diapause) have more number of mature eggs than either of those diapausing for short period (23-57 days) or those of longer diapause (90-165). &nbsp; Key words: Microplitis rufiventris, mass rearing conditions, obligate diapause, diapause intensity, generation effect, fecundity.

Effects of Pre-Diapause Temperature and Body Weight on the Diapause Intensity of the Overwintering Generation of Bactrocera minax (Diptera: Tephritidae).

The Chinese citrus fruit fly, Bactrocera minax (Enderlein), is an economically important pest of citrus. The fly has an obligatory pupal diapause in soil from November to March. However, techniques for predicting or determining the emergence of the adult have, thus far, not been well documented. In this study, we investigated the effects of different pre-diapause temperatures (8, 12, 16, and 20°C) and pupal body weight (five groups according to pupal weight: G-58, 55.0–61.0 mg; G-68, 65–71 mg; G-78, 75–81 mg; G-88, 85–91 mg; G-95, 92–98 mg) on pupal period (the indicator of diapause intensity). The pupal period of B. minax larvae pupated at 8°C was 193.41 d, which was significantly shorter than that of larvae incubated at higher temperatures, suggesting that there was a lower diapause intensity for larvae pupated at lower pre-diapause temperatures. There were also significant differences in the pupal periods at different pupal body weights. The pupal period of G-58 was significantly shorter than that of the heavier groups (G-88 and G-95), and the pupal period increased with increasing pupal body weight in the five groups. Moreover, the pupal period of B. minax significantly and positively correlated to pupal body weight. These findings demonstrate that the pre-diapause temperature and pupal body weight are suitable indicators for predicting the pupal period of overwintering individuals, and the results of this study will contribute to the development of new and effective strategies for predicting the occurrence and population dynamics of B. minax adult.

Influence of oviposition timing on offspring diapause and growth in Monochamus alternatus endai (Coleoptera: Cerambycidae)

ABSTRACTMonochamus alternatus endai Makihara is the primary vector of pine wilt disease in Japan and South Korea. Larvae of this cerambycid beetle exhibit induced diapause in the final instar. To determine the effects of seasonal environments that female adults had experienced on the diapause, growth, and development of offspring, parents of two Japanese strains were allowed to oviposit on Pinus densiflora logs over 12 or 4 periods of 6 or 7 days each between June and September under natural outdoor conditions. The offspring were reared at 25°C under a photoperiod of LD 12:12 h or LD 16:8 h for 17 weeks, then larvae in diapause were chilled at 10°C under LD 12:12 h for 8 or 16 weeks and transferred to 25°C under LD 12:12 h. Though maternal environments did not affect the diapause of offspring, early seasonal environments that parents had experienced made the offspring larval and adult body masses larger than late seasonal environments. The proportion of larvae that terminated diapause was smaller after an 8-week chilling period than after a 16-week chilling period, indicating that sufficient time of chilling was needed to terminate diapause in the laboratory. The proportion of larvae that terminated diapause was smaller when having experienced LD 16:8 h before chilling treatment than when experienced LD 12:12 h, indicating that LD 16:8 h made the diapause more intense than LD 12:12 h. That suggests that pre-diapause larvae early in the summer induce intense diapause while those early in the autumn induce weak diapause.

Expression Analysis Reveals the Association of Several Genes with Pupal Diapause in Bactrocera minax (Diptera: Tephritidae).

The Chinese citrus fly, Bactrocera minax, is a devastating pest of citrus, which enters the obligatory diapause in overwintering pupae to resist harsh environmental conditions. However, little is known about the molecular mechanisms underlying pupal diapause. The previous transcriptomic analysis revealed that a large number of genes were regulated throughout the pupal stage. Of these genes, 12 and six ones that are remarkably up- and downregulated, respectively, specifically in intense diapause were manually screened out in present study. To validate the expression of these genes throughout the pupal stage, the quantitative real-time PCR (qRT-PCR) was conducted, and the genes displaying different expression patterns with those of previous study were excluded. Then, the expressions of remaining genes were compared between diapause-destined and non-diapause-destined pupae to reveal their association with diapause using qRT-PCR and semiquantitative PCR. Finally, five genes, TTLL3B, Cyp6a9, MSTA, Fru, and UC2, and two genes, KSPI and LYZ1, were demonstrated to be positively and negatively associated with diapause, respectively. These findings provide a solid foundation for the further investigation of molecular mechanisms underlying B. minax pupal diapause.

Effects of photoperiod and chilling on diapause induction, intensity and termination in Monochamus alternatus endai (Coleoptera: Cerambycidae)

ABSTRACTMonochamus alternatus Hope adults vector the pinewood nematode, which is the causal agent of pine wilt disease. Larvae of this cerambycid beetle enter diapause in the final instar. To determine the effects of photoperiod and temperature on diapause in the subspecies M. a. endai, which is native to Japan, and to predict the effects of invasion by M. a. alternatus, the exotic subspecies with facultative diapause, immature stages of two M. a. endai strains (collected at 36°48ʹN, 136°48ʹE; 34°24ʹN, 132°43ʹE) were reared in Pinus densiflora bolts at 25°C under one of three LD photoperiods: 8:16 h, 12:12 h, and 16:8 h for 17 weeks, then chilled at 10°C under LD 10:14 h for 8 or 16 weeks, and finally transferred to 25°C under LD 12:12 h. All larvae entered diapause regardless of photoperiod during the initial incubation, verifying obligate diapause. The 16-week chilling period caused a greater incidence of diapause termination (0.908) than the 8-week chilling period (0.083). Diapause termination was also affected by the photoperiod treatment before chilling, M. a. endai strain, and their interaction. There was a difference in the diapause intensity between strains. The intensity of diapause was affected by photoperiods before chilling. If a small number of M. a. alternatus invade Japan, the obligate diapause requirement in M. a. endai, which is a dominant trait that a previous study verified, would not cause the inter-subspecies hybridization to change the life cycle of M. alternatus and consequently the epidemic pattern of pine wilt disease in Japan.

Summer diapause intensity influenced by parental and offspring environmental conditions in the pest mite, Halotydeus destructor

The regulation of active and dormant stages of arthropods is critical for surviving unfavourable seasonal conditions, and for many species depends on the diapause intensity (DI). There is substantial information on diapause strategies of arthropods under winter conditions; however, most cases of summer diapause are poorly understood despite its importance in most geographic regions of the world. Here we show how complex interactions with the environment drive DI involving multiple summer diapause forms of the mite Halotydeus destructor. This invasive pest in Australia is only active in cooler months but enters diapause at the egg stage which can survive hot and dry summer conditions. Recent research points to two forms of diapause egg, a typical form with a thick chorion and a cryptic form without this chorion which is morphologically similar to non-diapause eggs. Compared with typical diapause eggs which are produced in late spring, cryptic diapause eggs could be produced together with non-diapause eggs earlier in the season with relatively cooler temperatures and shorter daylength, reflecting an advanced bet-hedging strategy. Fitness trade-offs in this strategy are investigated in this study as variability of DI of the typical and cryptic diapause forms under different environmental factors for incubating diapause eggs (temperature) and rearing parental mites (different daylength, temperature and soil moisture). With the exception of daylength, all factors impacted hatchability of diapause eggs. Higher mortality of cryptic diapause eggs indicated relatively shallower DI than typical diapause eggs, likely reflecting a fitness penalty of this bet-hedging strategy under some conditions. Hatchability of cryptic diapause eggs revealed thermal and moisture stresses have opposite and complementary effects between parental and filial generations. Although DI of filial eggs decreased in hot and dry summer conditions, parental mites reared in hotter and drier conditions increased the DI of offspring. A bet-hedging strategy involving cryptic diapause might be replaced by typical diapause under consistently stressful conditions because of higher survival, regardless of additional production costs that might be required. These findings highlight a complex set of plastic responses to summer conditions in H. destructor that undoubtedly contribute to the success of this invasive pest under a range of environments.

Genomic Differentiation during Speciation-with-Gene-Flow: Comparing Geographic and Host-Related Variation in Divergent Life History Adaptation in Rhagoletis pomonella.

A major goal of evolutionary biology is to understand how variation within populations gets partitioned into differences between reproductively isolated species. Here, we examine the degree to which diapause life history timing, a critical adaptation promoting population divergence, explains geographic and host-related genetic variation in ancestral hawthorn and recently derived apple-infesting races of Rhagoletis pomonella. Our strategy involved combining experiments on two different aspects of diapause (initial diapause intensity and adult eclosion time) with a geographic survey of genomic variation across four sites where apple and hawthorn flies co-occur from north to south in the Midwestern USA. The results demonstrated that the majority of the genome showing significant geographic and host-related variation can be accounted for by initial diapause intensity and eclosion time. Local genomic differences between sympatric apple and hawthorn flies were subsumed within broader geographic clines; allele frequency differences within the races across the Midwest were two to three-fold greater than those between the races in sympatry. As a result, sympatric apple and hawthorn populations displayed more limited genomic clustering compared to geographic populations within the races. The findings suggest that with reduced gene flow and increased selection on diapause equivalent to that seen between geographic sites, the host races may be recognized as different genotypic entities in sympatry, and perhaps species, a hypothesis requiring future genomic analysis of related sibling species to R. pomonella to test. Our findings concerning the way selection and geography interplay could be of broad significance for many cases of earlier stages of divergence-with-gene flow, including (1) where only modest increases in geographic isolation and the strength of selection may greatly impact genetic coupling and (2) the dynamics of how spatial and temporal standing variation is extracted by selection to generate differences between new and discrete units of biodiversity.

R/K-like trade-off and voltinism discreteness: The implication to allochronic speciation in the fall webworm, Hyphantria cunea complex (Arctiidae).

North America has distinct types of Hyphantria moths (Arctiidae) characterized by red (RD)‐ and black (BL)‐headed larvae, of which the taxonomic status is unresolved. Genetic divergence of 26 populations, based on 710 bp of the mtCOI sequence, showed two phylogenetic lineages, which could not be connected in the haplotype network with 95% confidence. The two lineages are separated by 3.1% sequence divergence and should be considered for full species status. The estimated split occurred 1.2–1.6 million years ago. The range of the RD type covered most of the continent, whereas that of the BL type was limited to eastern deciduous forests. Several biological characteristics were differentiated in the zone of cohabitation where BL had more annual generations than RD. Spring emergence of BL precedes that of RD in the field by at least 1 month, because the diapause in BL was shallow, whereas it was deep in RD. Voltinism requires discreteness of numbers, which functions as a sink of hybrids between the two parental lines that have distinct but equally adaptive reproductive strategies; BL may be more r‐strategist‐like and RD more K‐strategist‐like, because fast‐developing BL has multivoltine life cycle, investing less silk proteins as the round‐the‐clock feeder, and slow‐developing RD univoltine one investing more silk as the nocturnal feeder. Also, intensity of diapause, deep in RD and weak in BL, was grossly different, which may enforce segregation of spring adults. Allochronic speciation avoiding coincidental occurrence of adult stages is therefore the most likely scenario. Because the adults never meet in nature, large morphological differentiation is not required.

A test of genomic modularity among life-history adaptations promoting speciation with gene flow.

Speciation with gene flow may require adaptive divergence of multiple traits to generate strong ecologically based reproductive isolation. Extensive negative pleiotropy or physical linkage of genes in the wrong phase affecting these diverging traits may therefore hinder speciation, while genetic independence or "modularity" among phenotypic traits may reduce constraints and facilitate divergence. Here, we test whether the genetics underlying two components of diapause life history, initial diapause intensity and diapause termination timing, constrain differentiation between sympatric hawthorn and apple-infesting host races of the fly Rhagoletis pomonella through analysis of 10,256 SNPs measured via genotyping-by-sequencing (GBS). Loci genetically associated with diapause termination timing were mainly observed for SNPs mapping to chromosomes 1-3 in the genome, most notably for SNPs displaying higher levels of linkage disequilibrium (LD), likely due to inversions. In contrast, selection on initial diapause intensity affected loci on all five major chromosomes of the genome, specifically those showing low levels of LD. This lack of overlap in genetically associated loci suggests that the two diapause phenotypes are largely modular. On chromosome 2, however, intermediate level LD loci and a subgroup of high LD loci displayed significant negative relationships between initial diapause intensity and diapause termination time. These gene regions on chromosome 2 therefore affected both traits, while most regions were largely independent. Moreover, loci associated with both measured traits also tended to exhibit highly divergent allele frequencies between the host races. Thus, the presence of nonoverlapping genetic modules likely facilitates simultaneous, adaptive divergence for the measured life-history components.

Geographic Variation of Diapause Induction and Termination in the Spider Mite Tetrαnychus urticαe: Α mini-review

In eight strains of the spider mite Tetranychus urticae Koch (Acari: Tetranychidae), originating from different localities in Europe, the critical daylength for diapause induction and termination was almost the same in each strain but varied with the latitudinal origin of the strains; critical daylength was shorter in strains originating from lower latitudes and longer in those from higher latitudes. Diapause intensity, measured as the period of chilling required for diapause termination under a short day photoperiod (LD 10:14) and 19°C, again varied with the latitudinal origin of each strain, being higher the more northern the origin of the strain. An exception were two mountain strains which showed a longer critical daylength and a deeper diapause than expected on the basis of their latitudinal origin. The number of long-day (LD 17:7) cycles required for 50% diapause termination after a certain period of chilling was higher in the northern and lower in the southern strains. These results indicate that geographic strains of T urticae may differ considerably in their diapause attributes, which may be explained as an adaptation to local climatic conditions. The great plasticity of the diapause response may, among other factors, have been responsible for the wide distribution of this mite species.

A year-round study on metabolic enzymes and body composition of the Arctic copepod Calanus glacialis: implications for the timing and intensity of diapause

Knowledge on the capability of zooplankton to adapt to the rapidly changing environmental conditions in the Arctic is crucial to predict future ecosystem processes. The key species on the Arctic shelf, the calanoid copepod Calanus glacialis, grows and accumulates lipid reserves in spring and summer in surface waters. The winter is spent in dormancy in deeper water layers with low metabolic activity. As timing and intensity of metabolic changes have been poorly investigated, our study aims to characterize the physiology of C. glacialis over an entire year, from July 2012 to July 2013. We followed anabolic and catabolic enzyme activities and the biochemical composition of this species, taking depth-stratified samples once a month in Billefjorden, a high-Arctic sill fjord. A large part of the population had migrated to depths >100 m by July 2012. Only thereafter, anabolic activities decreased slowly, suggesting that low metabolism is related to ceased feeding rather than to endogenous regulation. During overwintering, anabolic enzyme activities were reduced by half as compared to peak activities in spring. The biochemical composition of the copepods changed little from July to December. Then, the lipid catabolic activity increased and the lipid content decreased, likely fuelling moulting and gonad maturation. The protein content did not change significantly during winter, suggesting that proteins are not much catabolized during that time. The relatively high metabolic activity in C. glacialis in winter suggests that this species is not entering a true diapause and should thus be able to respond flexible to changing environmental conditions.

Energy and lipid metabolism during direct and diapause development in a pierid butterfly.

Diapause is a fundamental component of the life cycle in the majority of insects living in environments characterized by strong seasonality. The present study addresses poorly understood associations and trade-offs between endogenous diapause duration, thermal sensitivity of development, energetic cost of development and cold tolerance. Diapause intensity, metabolic rate trajectories and lipid profiles of directly developing and diapausing animals were studied using pupae and adults of Pieris napi butterflies from a population in which endogenous diapause has been well studied. Endogenous diapause was terminated after 3 months and termination required chilling. Metabolic and post-diapause development rates increased with diapause duration, while the metabolic cost of post-diapause development decreased, indicating that once diapause is terminated, development proceeds at a low rate even at low temperature. Diapausing pupae had larger lipid stores than the directly developing pupae, and lipids constituted the primary energy source during diapause. However, during diapause, lipid stores did not decrease. Thus, despite lipid catabolism meeting the low energy costs of the diapausing pupae, primary lipid store utilization did not occur until the onset of growth and metamorphosis in spring. In line with this finding, diapausing pupae contained low amounts of mitochondria-derived cardiolipins, which suggests a low capacity for fatty acid β-oxidation. While ontogenic development had a large effect on lipid and fatty acid profiles, only small changes in these were seen during diapause. The data therefore indicate that the diapause lipidomic phenotype is developed early, when pupae are still at high temperature, and retained until post-diapause development.

Photothermal control of larval diapause in the blowfly, Calliphora vicina R.-D. (Diptera, Calliphoridae) from the Lofoten Islands (Northern Norway)

Photothermal control of the rate of development and of larval diapause in the arctic population of Calliphora vicina from the Lofoten Islands (Northern Norway) was studied under the laboratory conditions. In the temperature range from 8 to 23°C, the rate of development of the studied population from egg to puparium can be well (r = 0.95) approximated by linear regression. The sum of effective temperatures was about 160 degree-days with the lower development threshold of 4.5 ± 0.6°C. The maternal photoperiodic response induced the larval diapause in a significant fraction of individuals only when larvae developed at temperatures of 4–10°C; relatively high proportion of diapausing larvae was recorded even in the progeny of females that were kept under long day conditions. The intensity of the larval diapause was very variable, which was manifested in the separation of diapausing larvae into two fractions differing in the time of pupariation during both spontaneous and induced reactivation.

Effects of environmental factors on the regulation of egg diapause in the walking-stick insect, Ramulus irregulariterdentatus (Phasmatodea: Phasmatidae)

Environmental factors that regulate egg diapause in the walking-stick insect, Ramulus irregulariterdentatus, were studied. Insects collected in fields at Okayama, Japan soon after hatching in spring were reared under constant laboratory conditions. After adult emergence, their eggs were placed outdoors or incubated under constant laboratory conditions. Most eggs kept at 15 or 20&deg;C hatched more than 100 days after oviposition. At 25&deg;C, however, many eggs remained in diapause throughout the experimental period although a small number hatched slightly earlier than those kept at the lower temperature. The maternal photoperiod affected the egg period at 20 and 25&deg;C; eggs from females reared under short day conditions hatched significantly earlier. A low temperature of 10&deg;C shortens diapause development, whereas 5&deg;C did not have a clear effect on time to hatching. When eggs of females reared under long day conditions were placed outdoors from June to early July, most hatched the next spring. On the other hand, eggs of females reared under short day conditions hatched before winter if they were laid before mid-July. The short day eggs laid in August and September hatched successfully the following April, and there was a positive correlation between the date of oviposition and time of hatching. It is concluded that low temperatures in mid-winter do not terminate diapause development. The results revealed that both maternal regulation of diapause intensity and the rate of diapause development play an important role in maintaining a monovoltine life cycle in Ramulus irregulariterdentatus.