Effects of parental diet on Mormon cricket egg diapause, embryonic development rate, and periodic outbreaks.

Transgenerational phenotypic modification can alter organismal fitness, population demographics, and community interactions. For ectotherms, both dietary composition and temperature have important effects on organismal fitness, but they are rarely investigated together. Mormon crickets Anabrus simplex are capable of diapausing as eggs in the soil for multiple years with duration largely dependent on cumulative heat units or degree days. Because Mormon crickets can be abundant in the landscape in one year and disappear suddenly the next, I asked: does parental nutrition affect the duration of egg diapause? Beginning in the ultimate nymphal instar, Mormon crickets were fed a diet high in protein, one equal in protein to carbohydrate, or a diet high in carbohydrates and the time for eggs to develop after they were laid was measured. If parental nutrition affects temperature-sensitive egg diapause, then that change in sensitivity to temperature might also alter the relationship between embryonic development rate and temperature. I asked: does parental nutrition affect embryonic development rate as a function of temperature? To this end, I manipulated densities of Mormon cricket nymphs and protein-rich prey (grasshoppers) in field cages, collected eggs from the adult Mormon crickets, and measured the optimal temperature, maximum development rate, and thermal breadth for embryonic development of the offspring. I found that Mormon crickets fed a high protein diet laid eggs with shorter diapause. Consistent with this long-term result, those housed with the most grasshoppers to eat laid eggs that had the fastest maximum development rate, whereas those without grasshoppers laid eggs with slower maximum developmental rates but the broadest thermal breadth. Eggs from Mormon crickets housed with intermediate levels of grasshopper densities had a decline in peak development rate with an increase in density. In addition, Mormon crickets housed with more conspecifics laid eggs with faster development rates, whereas thermal breadth and the temperature optima were not affected by cricket density. As predicted, Mormon cricket diets significantly affected egg diapause and development rates. Contrary to expectations based on observed changes in diet preferences during a Mormon cricket outbreak, Mormon crickets fed high protein diets laid eggs with significantly shorter egg diapause and significantly faster egg development rates. Interestingly, doubling of Mormon cricket density caused eggs to develop in nearly half the time. This latter result indicates that Mormon cricket aggregations promote rapid development of progeny. Moreover, the tight, linear structure of migratory bands in which females intermittently stop to lay eggs assures that the progeny hatch and develop in dense cohorts. In this manner, the banding behavior might carry-over into subsequent generations as long as cohorts are dense and protein is available. With band thinning or protein restriction, females spread their bet-hedging and progeny remain longer as eggs in the soil.

Metabolic Rate Suppression and Maintenance of Flight Muscle Metabolic Capacity during Diapause in Bumble Bee (Bombus impatiens) Queens.

AbstractThe common eastern bumble bee (Bombus impatiens) queens endure cold winter months by entering a diapause state. During this overwintering period, these animals use stored energy reserves while maintaining a low metabolic rate. This study investigates changes in the metabolic rate of bumble bee queens during diapause-like laboratory conditions and the potential reorganization of the flight muscle metabolic properties during this period. We first confirmed the hypometabolic state of queens during diapause in the laboratory, which lowered their resting metabolic rate to less than 5% of normal resting values. Body mass decreased during diapause, body composition changed where carbohydrates decreased initially, and later protein declined, with a similar trend for lipid content. Using cellular respirometry, we determined the capacity of the flight muscle cells of bumble bee queens to use various metabolic fuels and whether this capacity changes during the progression of diapause to favor stored lipid-derived substrates. Queens showed a low capacity to oxidize the amino acid proline, compared with workers, and their capacity to oxidize all metabolic substrates did not change during a 4-mo diapause period in the laboratory. We also show no detectable ability to oxidize fatty acid by flight muscle mitochondria in this species. The metabolic properties of flight muscle tissue were further characterized using metabolic enzyme activity profiles showing little change during diapause, indicating that profound metabolic suppression is induced without major changes in muscle metabolic phenotypes. Overall, B. impatiens queens undergo diapause while maintaining flight muscle capacity under the conditions used.

Altered circadian rhythm, sleep, and rhodopsin 7-dependent shade preference during diapause in Drosophila melanogaster.

To survive adverse environments, many animals enter a dormant state such as hibernation, dauer, or diapause. Various Drosophila species undergo adult reproductive diapause in response to cool temperatures and/or short day-length. While flies are less active during diapause, it is unclear how adverse environmental conditions affect circadian rhythms and sleep. Here we show that in diapause-inducing cool temperatures, Drosophila melanogaster exhibit altered circadian activity profiles, including severely reduced morning activity and an advanced evening activity peak. Consequently, the flies have a single activity peak at a time similar to when nondiapausing flies take a siesta. Temperatures ≤15 °C, rather than photoperiod, primarily drive this behavior. At cool temperatures, flies rapidly enter a deep-sleep state that lacks the sleep cycles of flies at higher temperatures and require high levels of stimulation for arousal. Furthermore, we show that at 25 °C, flies prefer to siesta in the shade, a preference that is virtually eliminated at 10 °C. Resting in the shade is driven by an aversion to blue light that is sensed by Rhodopsin 7 outside of the eyes. Flies at 10 °C show neuronal markers of elevated sleep pressure, including increased expression of Bruchpilot and elevated Ca2+ in the R5 ellipsoid body neurons. Therefore, sleep pressure might overcome blue light aversion. Thus, at the same temperatures that cause reproductive arrest, preserve germline stem cells, and extend lifespan, D. melanogaster are prone to deep sleep and exhibit dramatically altered, yet rhythmic, daily activity patterns.

Impact of climate change on the reproductive diapause and voltinism of the carrot weevil, Listronotus oregonensis.

The impacts of climate change on the development of insects are of great concern due to potential alterations in population dynamics and pest pressure. The carrot weevil, Listronotus oregonensis, is a major agricultural pest, and its development is influenced by temperature and photoperiod. In this study, our aim was to investigate the impact of temperature increases on the voltinism and reproductive diapause of the carrot weevil under field conditions and bioclimatic models. Field observations were conducted over two growing seasons using structures that allowed for temperature increases. The developmental stages of the carrot weevil, including female reproductive status, oviposition and larval stage, were monitored weekly to measure the proportion of individuals undergoing an additional generation. Concurrently, bioclimatic models were used to simulate the probability of a second generation under current (1981-2010) and future (2041-2070) climates, considering a lower and a higher change in emission scenarios. Results showed that rising temperatures led to an increase in the proportion of carrot weevils undergoing inhibition of the reproductive diapause and a higher number of eggs laid in the field. The models indicated a substantial rise in the probability of a second generation developing, from 24% to 37% to 62%-99% under current and future climates, respectively. These findings demonstrate the potential for significant alterations in carrot weevil population dynamics, resulting in increased pest pressure on crops. Further research is needed to fully understand the implications of these findings and to develop effective adaptation measures to mitigate the negative impacts of global warming on insect populations and agriculture.

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.

Prolonged diapause in Mormon crickets: Embryonic responses to three measures of time.

Mormon cricket eggs can remain diapausing in soil for multiple years without forming an embryo. I investigated whether embryonic development was dependent on the number of annual cycles since the egg was laid, duration of the summer period (forcing), or duration of the winter period (chilling). Male and female Mormon crickets collected in Arizona and Wyoming were paired in the lab. For each mating pair, sibling eggs were incubated 12 weeks, eggs with fully developed embryos removed, and the remaining eggs were split evenly among three treatments: a long cold period and a long warm period; a short cold period and a long warm period; and a short cold period and a short warm period, which respectively completed 2 annual cycles, 3 cycles, and 4 cycles in 60 calendar weeks. In each cycle over nine years, developed eggs and eggs that appeared inviable were counted and removed. For each mating pair, I used survival analyses to test for differences in 1) the number of annual cycles, 2) the warm period duration, and 3) the cold period duration required for the embryos to develop. For eight of 11 mating pairs, one of the three factors was not excluded as a determinant of the phenology of embryonic development. Duration of the warm period was not rejected in seven of 11 cases. Duration of the warm period required for 50 % of the eggs to develop ranged from 84 to 144 weeks. In one case from Arizona, the duration of the cold period was the only factor not rejected. Median chill time was 60 weeks, which is also more than one year. Despite this exception, I conclude that duration of the warm period is typically the factor that determines timing of embryonic development for Mormon crickets. For these two high elevation populations, median forcing or chilling exceeded one year.

Neuropeptidergic regulation of insect diapause by the circadian clock.

Diapause is an endocrine-mediated strategy used by insects to survive seasons of adverse environmental conditions. Insects living in temperate zones are regularly exposed to such conditions in the form of winter. To survive winter, they must prepare for it long before it arrives. A reliable indicator of impending winter is the shortening of day length. To measure day length, insects need their circadian clock as internal time reference. In this article, I provide an overview of the current state of knowledge on the neuropeptides that link the clock to the diapause inducing hormonal brain centers.

Management of inorganic elements by overwintering physiology of cold hardy larvae of European corn borer (Ostrinia nubilalis, Hbn.).

The European corn borer (Ostrinia nubilalis, Hbn.), enters diapause, a strategy characterized by arrest of development and reproduction, reduction of metabolic rate and the emergence of increased resistance to challenging seasonal conditions as low sub-zero winter temperatures. The aim of this study was to investigate the potential role of inorganic elements in the ecophysiology of O. nubilalis, analysing their content in the whole body, hemolymph and fat body, both metabolically active, non-diapausing and overwintering diapausing larvae by ICP-OES spectrometer following the US EPA method 200.7:2001. O nubilalis as many phytophagous lepidopteran species maintain a very low extracellular sodium concentration and has potassium as dominant cation in hemolymph of their larvae. Changes in hemolymph and the whole body sodium content occur already at the onset of diapause (when the mean environmental temperatures are still high above 0 ºC) and remain stable during the time course of diapause when larvae of this species cope with sub-zero temperatures, it seems that sodium content regulation is rather a part of diapausing program than the direct effect of exposure to low temperatures. Compared to non-diapausing O. nubilalis larvae, potassium levels are much higher in the whole body and fat body of diapausing larvae and substantially increase approaching the end of diapause. The concentration of Ca, Mg, P and S differed in the whole body, hemolymph and fat body between non-diapausing and diapausing larvae without a unique trend during diapause, except an increase in their contents at the end of diapause.

The costs of overwintering in paper wasps (Polistes dominula and Polistes gallicus): the use of energy stores.

Overwintering insects are facing energetic challenges because of food shortage, low temperature, and desiccation stress. Paper wasps of the genus Polistes overwinter as mated adults (gynes) in hibernacula protecting them from predation, snow, and rain but barely from low environmental temperature. In different climates, they face differing overwintering temperature regimes, and therefore they may differ in their energy use. We investigated how much of energy resources built up until autumn is used during diapause dormancy in natural hibernacula by measuring lipid, glycogen, and free carbohydrate content in autumn and early spring in Polistes dominula from temperate European (Austrian) and warm Mediterranean (Italian) climate and Polistes gallicus from Mediterranean climate. Winter energy consumption amounted to ~ 339 and ~ 310 J per wasp in the Austrian and Italian Polistes dominula populations. The smaller Italian Polistes gallicus consumed ~ 247 J. This amounts to 2.62, 2.35, and 1.79 J per day. Of this, the energy demand was mainly fuelled by lipids (84%, 93%, and 90%, respectively), but glycogen stores contributed also considerably (16%, 6%, and 9%). Free carbohydrates decreased only by 0.7%, 1%, and 0.8%. While fat stores seem still sufficient in spring, the wasps depleted most of their carbohydrates. The energy reserves of 396, 400, and 147 J per wasp remaining in spring in the three populations seem sufficient to fuel rest or simple brood care activities for a whole summer but restrict foraging flights to a few hours (~ 3.5-6 h). Results suggest that energy supply might become challenging in expected future climate scenarios.

Genetic constraints in genes exhibiting splicing plasticity in facultative diapause.

Phenotypic plasticity is produced and maintained by processes regulating the transcriptome. While differential gene expression is among the most important of these processes, relatively little is known about other sources of transcriptional variation. Previous work suggests that alternative splicing plays an extensive and functionally unique role in transcriptional plasticity, though plastically spliced genes may be more constrained than the remainder of expressed genes. In this study, we explore the relationship between expression and splicing plasticity, along with the genetic diversity in those genes, in an ecologically consequential polyphenism: facultative diapause. Using 96 samples spread over two tissues and 10 timepoints, we compare the extent of differential splicing and expression between diapausing and direct developing pupae of the butterfly Pieris napi. Splicing differs strongly between diapausing and direct developing trajectories but alters a smaller and functionally unique set of genes compared to differential expression. We further test the hypothesis that among these expressed loci, plastically spliced genes are likely to experience the strongest purifying selection to maintain seasonally plastic phenotypes. Genes with unique transcriptional changes through diapause consistently had the lowest nucleotide diversity, and this effect was consistently stronger among genes that were differentially spliced compared to those with just differential expression through diapause. Further, the strength of negative selection was higher in the population expressing diapause every generation. Our results suggest that maintenance of the molecular mechanisms involved in diapause progression, including post-transcriptional modifications, are highly conserved and likely to experience genetic constraints, especially in northern populations of P. napi.

Significance of the clock gene period in photoperiodism in larval development and production of diapause eggs in the silkworm Bombyx mori.

Many insects living in seasonal environments sense seasonal changes from photoperiod and appropriately regulate their development and physiological activities. Genetic researches have indicated the importance of a circadian clock system in photoperiodic time-measurement for photoperiodic regulations. However, most previous studies have focused on the effects on a single photoperiodic phenotype, without elucidating whether the circadian clock is involved in the core photoperiodic mechanism or only in the production of one target phenotype, such as diapause. Here, we focused on two different phenotypes in a bivoltine Kosetsu strain of the silkworm Bombyx mori, namely, embryonic diapause and larval development, and examined their photoperiodic responses and relationship to the circadian clock gene period. Photoperiod during the larval stage clearly influenced the induction of embryonic diapause and duration of larval development in the Kosetsu strain; short-day exposure leaded to the production of diapause eggs and shortened the larval duration. Genetic knockout of period inhibited the short-day-induced embryonic diapause. Conversely, in the period-knockout silkworms, the larval duration was shortened, but the photoperiodic difference was maintained. In conclusion, our results indicate that the period gene is not causally involved in the photoperiodic response of larval development, while that is essential for the short-day-induced embryonic diapause.