Winter Diapause Research Articles

Effects of Irradiation on Biology and Mating Behaviour of Wild Males of Brown Marmorated Stink Bug Using a 6 MV Medical Linear Accelerator

The brown marmorated stink bug, Halyomorpha halys, is a pentatomid bug of Eastern Asian origin that became an economically relevant pest in the Eurasian and American continents. Management of this species is limited to use of chemical insecticides: an inefficient method due to the strong adaptability of the target pest. The sterile insect technique (SIT) is potentially a valid tactic in the search for nontoxic alternatives. In this work, we investigated the suitability of mass-trapped overwintering males, collected during the aggregation phase before the winter diapause, for their release as competitive sterile males in an SIT programme. Differently from previous studies, irradiation was applied with a linear accelerator device that produced high-energy photons. Following a similar scientific protocol with newly emerged irradiated males, the effects of X-ray irradiation on physiological parameters (longevity, fecundity and fertility) were assessed. In addition, behavioural bioassays were carried out in no-choice conditions to evaluate if irradiation interferes with mating processes. The results are very encouraging; the effects of the irradiation at 32 Gy did not differ from the controls in the longevity or fecundity of the exposed overwintering adults. The hatching rate of the eggs laid by the fertile females that had mated with the irradiated males was less than 5%. The results of behavioural bioassays showed that the irradiation did not cause a significant impact on the quality of the sterile males. More research is warranted to evaluate the mating competitiveness of sterile males in semi-field and field conditions.

Prolonged heat stress during winter diapause downregulates gene expression of attacin and gloverin, two antimicrobial peptides in the European corn borer, Ostrinia nubilalis (Hübner)

Prolonged heat stress during winter diapause downregulates gene expression of attacin and gloverin, two antimicrobial peptides in the European corn borer, Ostrinia nubilalis (Hübner)

The Invasive Caucasian Populations of the Brown Marmorated Stink Bug Halyomorpha halys (Hemiptera: Heteroptera: Pentatomidae) Rapidly Adapt Their Ecophysiological Traits to the Local Environmental Conditions

The ability to rapidly adapt to new environmental conditions is a crucial prerequisite for the wide-scale invasion of pests or intentional introduction of beneficial insects. A photoperiodically induced facultative winter diapause is an important adaptation ensuring synchronization of insect development and reproduction with the local seasonal dynamics of environmental factors. We conducted a laboratory study aimed to compare photoperiodic responses of two invasive Caucasian populations of the brown marmorated stink bug Halyomorpha halys (Hemiptera: Heteroptera: Pentatomidae), which recently invaded neighboring regions with subtropical (Sukhum, Abkhazia) and temperate (Abinsk, Russia) climates. Under the temperature of 25 °C and the near-critical photoperiods of L:D = 15:9 h and 15.5:8.5 h, the population from Abinsk showed a slower pre-adult development and a stronger tendency to enter winter adult (reproductive) diapause compared to the population from Sukhum. This finding agreed with the difference between the local dynamics of the autumnal temperature decrease. Similar adaptive interpopulation differences in the patterns of diapause-inducing responses are known in other insect species but our finding is distinguished by a very short adaptation time: H. halys was first recorded in Sukhum in 2015 and in Abinsk in 2018. Thus, the differences between the compared populations might have evolved over a relatively short span of several years.

Warming increases survival and asexual fitness in a facultatively sexual freshwater cnidarian with winter diapause.

Temperature is a key abiotic factor controlling population dynamics. In facultatively sexual animals inhabiting the temperate zone, temperature can regulate the switch between asexual and sexual modes of reproduction, initiates growth or dormancy, and acts together with photoperiod to mediate seasonal physiological transitions. Increasing temperature due to recent global warming is likely to disrupt population dynamics of facultatively sexual animals because of the strong temperature dependence of multiple fitness components. However, the fitness consequences of warming in these animals are still poorly understood. This is unfortunate since facultatively sexual animals-through their ability for asexual reproduction resulting in quick population growth and sexual reproduction enabling long-term persistence-are key components of freshwater ecosystems. Here, I studied the fitness effects of warming in Hydra oligactis, a freshwater cnidarian that reproduces asexually throughout most of the year but switches to sexual reproduction under decreasing temperatures. I exposed hydra polyps to a simulated short summer heatwave or long-term elevated winter temperature. Since sexual development in this species is dependent on low temperature, I predicted reduced sexual investment (gonad production) and elevated asexual fitness (budding) in polyps exposed to higher temperatures. The results show a complex effect of warming on sexual fitness: While gonad number decreased in response to warming, both male and female polyps exposed to high winter temperature were capable of multiple rounds of gamete production. Asexual reproduction and survival rate, on the contrary, clearly increased in response to higher temperature, especially in males. These results predict increased population growth of H. oligactis in temperate freshwater habitats, which will likely affect the population dynamics of its main prey (freshwater zooplankton), and through that, the whole aquatic ecosystem.

Juvenile hormone identification in the cabbage bug Eurydema rugosa

Juvenile hormone (JH) plays a pivotal role in almost every aspect of insect development and reproduction. The chemical structure of the JH in heteropteran species has long remained elusive until methyl (2R,3S,10R)-2,3;10,11-bisepoxyfarnesoate, commonly named as juvenile hormone III skipped bisepoxide (JHSB3), was isolated from Plautia stali (Hemiptera: Heteroptera: Pentatomidae). Recently, several groups reported the presence of JHSB3 in other heteropteran species. However, most of the studies paid no attention to the determination of the relative and absolute structure of the JH. In this study, we investigated the JH of the cabbage bug Eurydema rugosa (Hemiptera: Heteroptera: Pentatomidae), known as a pest for wild and cultivated crucifers. JHSB3 was detected in the hexane extract from the corpus allatum (CA) product using a chiral ultraperformance liquid chromatography-tandem mass spectrometer (UPLC-MS/MS) which can inform the absolute stereochemistry of the JH. Its stereoisomers were not detected. Topical application of the synthetic JHSB3 to the last instar nymphs inhibited their metamorphosis and induced nymphal-type colouration of the dorsal abdomen in a dose-dependent manner. Additionally, the topical application of JHSB3 effectively terminated summer and winter diapauses in females. These results indicate that the JH of E. rugosa is JHSB3. Although individuals in summer and winter diapauses are physiologically distinct in E. rugosa, the results suggest that the physiological differences between these diapauses are based, not on the responsiveness to JH, but on the processes governing activation of the CA or on its upstream cascades.

The Effect of Pollen Diet Composition and Quantity on Diapause Survival and Performance in an Annual Pollinator (Bombus Impatiens).

Most pollination services are provided by annual bees that go through a winter diapause, during which they are exposed to extreme temperatures, pathogens, and starvation. The ability of bees to successfully face these stressors during diapause and subsequently initiate a nest depends on their overall nutritional state and an adequate preparatory diet. Here, we used queens of the common eastern bumble bee, Bombus impatiens, to examine how pollen diets varying in their protein to lipid ratio and total nutrient amounts affected queen performance during and after diapause. We compared diapause survival and reproductive performance post-diapause across different diets and found that queen survival was highest when pollen had a nutritional ratio of approximately 5:1 (protein to lipid). This diet is significantly enriched in proteins compared to the pollen fed to bumble bees in the lab (1:1) or commonly available in agricultural landscapes. Altering the quantity of macronutrients within this ratio did not improve survival or performance. Our results emphasize the importance of adequate nutrition for diapause performance in bees with annual life cycles and the importance of providing annual bees with floral provisioning based on their individual nutritional targets.

Effects of soil humidity on respiration and frost resistance during winter diapause in the pine beauty moth, Panolis flammea

Abstract Panolis flammea is an important pest whose populations intermittently outbreak in Europe. The species overwinters as pharate moths in the pupal stage in soil. Details on the metabolic activity and cold hardiness of P. flammea during the overwintering period have not been published. Therefore, we assessed O2 consumption and the supercooling point (SCP) of P. flammea in late November, mid‐February, and mid‐March and the influence of soil humidity under fluctuating temperatures and with brief exposures to two increased temperature regimes (10 and 20°C). The respiration pattern indicated diapause termination in mid‐February. A threshold of 0.169 μl O2 h−1 mg−1 fresh weight indicated potentially activated pre‐emerging moths. Drought increased respiration rates. The SCP was lowest in mid‐February (−22.2°C) and was negatively correlated with pupal mass. The frost tolerance tended to increase with low substrate humidity, especially at the end of the overwintering period. Our results indicate that P. flammea requires more energy during dry and mild winters than in wet and cold winters. Winter diapause termination and post‐diapause development may therefore be accelerated as environmental warming and drought increase.

Tracking Asian tiger mosquito introductions in the Netherlands using Nextstrain

The Asian tiger mosquito Aedes albopictus is an undesirable invasive mosquito species that causes considerable nuisance through its biting behaviour, and has been proven to transmit more than 22 different viruses under laboratory conditions. Human-aided transportation, the capacity of winter diapause, and possibly global warming have contributed to the global invasion of Ae. albopictus. The species was found for the first time in the Netherlands in 2005, and since 2010 has been found introduced at many locations throughout the country. Elucidating the origin of these introduced mosquitoes could help the authorities on the planning and evaluation of the risk-based surveillance of Aedes invasive mosquitoes. This study aims to determine the genomic diversity of Ae. albopictus that is represented within and between collection sites with a database consisting of Ae. albopictus specimens from past introductions in the Netherlands, specimens from populations from other regions in the world, and data from specimens present in databases. In this study, complete mitochondrial genomes were sequenced, a recommended marker for phylogeography analysis of Ae. albopictus. Metadata is presented in a Nextstrain build containing 254 Ae. albopictus genomes up to October 2020. Overall, the phylogeny results of the Nextstrain build reveals a low mitogenomic diversity within Ae. albopictus. Genomic diversity of Ae. albopictus specimens found in the Netherlands fall within one main cluster which is hypothesised to represent the globally invasive strain of the species. Other organisations are stimulated to share data or materials for inclusion and improvement of the Nextstrain build, which can be accessed at https://nextstrain.nrcnvwa.nl/Aedes/20210728.

SPATIAL DISTRIBUTION AND REALIZATION OF THE LIFE CYCLE OF THE GROUND BEETLE PTEROSTICHUS ADSTRICTUS ESCHSCHOLTZ, 1823 (COLEOPTERA, CARABIDAE) IN THE KOMSOMOLSKY NATURE RESERVE, KHABAROVSKY KRAI

The spatial distribution of the ground beetle Pterostichus adstrictus Eschscholtz, 1823 in the Komsomolsky Nature Reserve on five forest permanent sample plots in 2017–2019 are examined. It is shown that the species in this protected area implements a one-year monovoltine life cycle with a spring-summer type of reproduction and imaginal diapause in winter.

Gregarine parasites are adapted to mosquito winter diapause

The Asian tiger mosquito Aedes albopictus is one of the most invasive species of mosquito. The prevalence of its apicomplexan gregarine parasite Ascogregarina taiwanensis is high in natural populations across both temperate and tropical regions. However, the parasite’s oocysts cannot colonize the insect host during winter, when the mosquito lays diapausing eggs. It is therefore unclear if the parasite can survive outside of its insect host during the cold season in temperate regions. Oocysts stored for 1 month at a low temperature (representative of the temperatures that occur during periods of mosquito diapause) were as infectious as fresh oocysts, but those stored for the same period of time at a higher temperature (representative of the temperatures that occur during periods of mosquito activity) were uninfectious. We therefore suggest that the parasite has evolved traits that maximize its maintenance during periods of mosquito dormancy, while traits that would enable its long term survival during periods of mosquito activity have not been selected for.

Characterization of Three Heat Shock Protein Genes in Pieris melete and Their Expression Patterns in Response to Temperature Stress and Pupal Diapause.

Simple SummaryPieris melete, a major pest of crucifers, undergoes obligatory diapause as pupae to survive unfavorable temperature extremes during hot summers and cold winters. Heat shock proteins 70 (Hsp70) participate in this process; however, little is known about the underlying changes in Hsp70 expression both during the summer and winter diapause. The study aimed to investigate expression patterns of Hsp70s (PmHsc70/PmHsp70a, b) in response to diapause and short-term temperature stresses. The results showed that the expression of PmHsc70 and PmHsp70b were upregulated both in summer and winter diapause. Heat shock significantly induced up-regulation of the three genes in both summer and winter diapause. In non-diapause pupae, none of the genes responded to cold or heat stress. Further, it was found that 39 °C for 30 min was the most sensitive heat stress condition for PmHsc70 expressions in summer diapause and all three genes’ expressions in winter diapause. During summer diapause, the expression of the genes was up-regulated in response to high temperature acclimation at 31 °C. Meanwhile, only PmHsp70a and PmHsp70b were up-regulated when acclimated to a low temperature of 4 °C in winter diapause. In conclusion, the current results indicate that PmHsp70s plays a crucial role during both summer and winter diapause, in response to temperature stresses; and our findings may contribute to the increasing knowledge on seasonal diapause adaption.Heat shock protein 70 genes participate in obligatory pupal diapause in Pieris melete to survive unfavorable conditions. In this study, three full-length cDNAs of PmHsc70, PmHsp70a and PmHsp70b were identified, and their expression patterns in response to diapause and short-term temperature stresses were investigated. Summer and winter diapause were induced in the pupae and non-diapause individuals were used as a control. The pupae from each diapause group were subjected to either hot or cold conditions and the expression levels of the HSP genes were measured. Our results showed that up-regulation of PmHsc70 and PmHsp70b were detected both in summer and winter diapause, but not for PmHsp70a. Under cold stress, PmHsp70a and PmHsp70b were upregulated in summer and winter diapause, while heat shock significantly induced upregulation of all three genes. In non-diapause pupae, none of the genes responded to cold or heat stress. Furthermore, we found that incubation at 39 °C for 30 min was the most sensitive heat stress condition for PmHsc70 expression in summer diapause. On the other hand, the same temperature was effective for PmHsc70, PmHsp70a, and PmHsp70b expression in winter diapause. During summer diapause, expression of all three genes was upregulated in response to high-temperature acclimation at 31 °C, but only PmHsp70a and PmHsp70b were upregulated when acclimated to a low temperature of 4 °C in winter diapause. These results suggest that the PmHsc70, PmHsp70a, and PmHsp70b respond differently to pupal diapause and temperature stress, and that PmHsc70 is more sensitive to heat shock than to cold stress.

A multi-year dormancy strategy in a cabbage beetle population in southeastern China.

The life cycle of the cabbage beetle Colaphellus bowringi in southeastern China is complex due to four options for adult development: summer diapause, winter diapause, prolonged diapsuse, and nondiapause. However, detailed information on the multi‐year emergence patterns of diapausing individuals in this beetle has not been documented. In this study, we monitored the adult emergence patterns of diapausing individuals and estimated the influence of the diapause‐inducing temperature and photoperiod on the incidence of prolonged diapause under seminatural conditions for several years. The duration of diapause for adults collected from the vegetable fields in different years varied from several months to 5 years. Approximately 25.9%–29.2% of individuals showed prolonged diapause (emergence more than 1 year after entering diapause) over the 5 years of observation. Furthermore, regardless of insect age, the emergence of diapausing adults from the soil always occurred between mid‐February and March in spring and between late August and mid‐October in autumn, when the host plants were available. The influence of diapause‐inducing temperatures (22, 25, and 28°C) combined with different photoperiods (L:D 12:12 h and L:D 14:10 h) on diapause duration was tested under seminatural conditions. Pairwise comparisons of diapause duration performed by the log‐rank test revealed that the low temperature of 22°C combined with the long photoperiod of L:D 14:10 h induced the longest diapause duration, whereas the low temperature of 22°C combined with the short photoperiod of L:D 12:12 h induced the highest proportion of prolonged diapause. This study indicates that C. bowringi adopts a multi‐year dormancy strategy to survive local environmental conditions and unpredictable risks.

Development of two common dragonfly species with diverging occupancy trends

The two sibling and syntopic odonate species Sympetrum striolatum and Sympetrum vulgatum are common and widespread in Central Europe. While S. striolatum has strong positive population trends, declines of S. vulgatum are observed. The aim of this study was to identify possible drivers of these diverging trends. We presumed that differences in egg development may lead to differences in survival until hatching. First, eggs laid in non-permanent or shrinking waterbodies may suffer of increasing drought periods. Second, differences in developement may cause increased size-mediated intraguild predation, a common cause of reduced survival in odonate larvae. Egg development time and hatching rates were recorded of eleven egg clutches of S. vulgatum and ten clutches of S. striolatum under simulated drought vs. water and direct vs. delayed development treatments. Hatching rates were reduced under drought conditions, and particularly so in S. vulgatum. We did not observe obligate winter diapause in any of the egg clutches. But, S. vulgatum varied widely in development between clutches, while the eggs of S. striolatum developed much faster and hatched highly synchronously. This would provide S. striolatum with a temporal advantage that may lead to a size-advantage over most S. vulgatum. We also found that S. vulgatum grew faster. Faster larval growth would only compensate for those S. vulgatum with fast egg development. The current population trends may be partly attributed to lowered survival of S. vulgatum under drought and by phenological and, thus, size benefits of S. striolatum.Implications for insect conservationOur results show that population dynamics of two closely related dragonfly species can be explained by climatically induced changes in their interactions. Understanding the causes and processes of behavioural changes resulting in differing population trends is fundamental for the protection of species.

Identification and up-regulation of three small heat shock proteins in summer and winter diapause in response to temperature stress in Pieris melete

Small heat shock proteins (sHSPs) are conserved proteins that play key roles in organismal adaptation to adversity stressors. However, little is known about sHSPs during summer diapause. Three sHSP genes: PmHSP19.5, PmHSP19.9, and PmHSP20.0 were identified and cloned from Pieris melete. Sequence alignment and phylogenetic analysis revealed that the three sHSPs have a typical, conserved α-crystallin domain. PmHSP19.5 and PmHSP20.0 were both upregulated in summer diapause (SD) and winter diapause (WD), compared to non-diapause (ND) pupae. All three sHSPs were upregulated and showed similar trends in response to thermal stress. The 0 °C chilling treatment slightly affected sHSP transcripts in ND pupae, whereas both PmHSP19.5 and PmHSP19.9 were upregulated and PmHSP20.0 was downregulated after chilling at 0 °C for 24–96 h in both SD and WD pupae. The transcripts of PmHSP19.5 and PmHSP19.9 were significantly induced at 31 °C for 30 d in SD and WD pupae. The PmHSP20.0 transcript gradually decreased during the SD and WD programs. This is the first time that sHSPs have been linked to both overwintering and summer diapause processes. These findings suggest that sHSPs are involved in both summer and winter diapause maintenance and play a possible key role in temperature stress.

CHARACTERISTICS OF THE DEVELOPMENT CYCLES OF DIURNAL LEPIDOPTERA (LEPIDOPTERA: HESPERIOIDEA, PAPILIONOIDEA) ON THE TERRITORY OF UDMURTIA

The typology of the life cycles of diurnal lepidoptera of Udmurtia is carried out using such ecological and biological indicators as the duration of generation development (number of years), the level of voltinism during the growing season, seasonal features of the imago flight, development stages at which the winter diapause occurs and its territorial confinement. On this basis, 24 types of cycles have been singled out. Species with annual development (96.7 %) and one generation during the growing season (76.4 %) are strongly predominant. According to the timing of flight, the bulavous are divided into 5 phenological groups, the proportion of which is as follows: spring - 13.8 %, pre-summer - 11.4 %, early-summer - 36.6 %, mid-summer - 35.8 %, late-summer - 2.4 %. Wintering of species is carried out at all stages of development with a predominance of larval diapause (61.5 %). Such species as Vanessa atalanta and V. cardui have the status of ex-regional migrants. The latter species is characterized by diapause-free polycyclic development throughout the year. In addition, the forms of facultative bivoltinism and summer diapause are considered.

Life cycle of the tick Ixodes ricinus (L.) (Acari: Ixodidae) in the North-West of Russia

The characters of the life cycle and demographic processes of Ixodes ricinus (L., 1758), (Acari: Ixodidae) populations in the North-West of Russia were studied using methods of the long-term observations of ticks in insets in the natural biotopes (2013–2018). It was found that in the North-West of Russia, I. ricinus had a long period of seasonal activity (from April until October) and a long calendar age. Larvae, nymphs, as well as adults of both sexes can remain active for up to 12–15 months when they meet with appropriate host and feed successfully. Up to 9±0.8% of adults can overwinter twice and remain active until the end of June of the third year of their life (23 months). The I. ricinus tick at every developmental stage forms a certain sub-group of population in August–September. Due to the highly prolonged life the larvae, nymphs and females of I. ricinus are able to feed, and the females can lay eggs during their entire active period from April till October. At the same time, larvae and nymphs that feed from April to June molt in the year of feeding, and their development proceeds according to a three-year scenario. The females feeding from April to June can produce offspring during the same season, thus forming a 3-year developmental cycle. By contrast, all individuals (larvae, nymphs and females) feeding from July to October molt to the next stage in August–September of the following year. Females feeding in July–September can lay eggs both in the year of feeding and after winter diapause. This leads to an increase in the duration of the basic 3-year developmental cycle and its complexity. If the ticks molted the next year after feeding only once in the life cycle, the whole life cycle would increase by a year. If they did it in every developmental stage, then the life cycle would increase by three years. Thus, the total duration of development of ticks of one generation may last from 3 to 6 years. Some adult ticks (not more than 9±0.8% in the population) can remain active until the end of June of the seventh season.

Predictive simulation of spring development of Cydalima perspectalis Walker in the South of Russia

The work presents the results of a study of the features of the seasonal development of a dangerous invasive insect the box tree moth, Cydalima perspectalis Walker, specialised on plants of the genus Buxus L. Models for forecasting the dates of the resumption of larvae activity in spring after the winter diapause, the first adult flights of the overwintered generation and the appearance of the first summer generation of larvae have been developed for the southern regions of Russia. The accuracy of the forecast of the larvae coming out of the winter diapause is 3-4 days, of the adult flights and the appearance of the first summer of larvae is 9-10 days. The developed models can be used as a tool to improve the control of this pest.

Ultrastructure of spherites in the midgut diverticula and Malpighian tubules of the harvestman Amilenus aurantiacus during the winter diapause

Amilenus aurantiacus overwinter in diapause, a natural starvation period, in hypogean habitats. The structure of spherites in the midgut diverticula (MD) and Malpighian tubules (MT) has been studied comparatively by light microscopy and TEM to detect eventual differences in mineral consumption in the beginning and at the end of the starvation period in these organs (MD and MT) associated with digestive processes. The chemical composition of spherites was examined by combining energy-dispersive X-ray spectroscopy (EDXS), electron energy-loss spectroscopy (EELS) and energy-filtered TEM (EFTEM). The structure of the spherites changed during overwintering in both organs. At the beginning of overwintering, the spherites were composed of densely packed concentric layers of electron-dense and electron-lucent material. In the middle and at the end of overwintering, the electron-lucent layers between the layers of material indicated the loss of some material. The chemical composition of the spherites changed only in the MD; at the beginning of overwintering, these contained Si, O, C and Fe, while later there was no more Fe. In contrast, spherites in the MT were composed of Si, O, C and Ca throughout overwintering. A less intensive exploitation of the MD spherites was probably due to complete cessation of digestive and other cell activity in this organ during the winter diapause; activity of the MT slowed, but continued removing the cell metabolites.

Conditions to Terminate Reproductive Diapause of a Univoltine Insect: Ceratapion basicorne (Coleoptera: Apionidae), a Biological Control Agent of Yellow Starthistle.

Ceratapion basicorne (Illiger) is a recently approved univoltine biological control agent that develops inside the rosette of yellow starthistle (Centaurea solstitialis L.), an invasive annual plant. Adult weevils normally emerge in early summer, and females are thought to be in reproductive diapause until the following spring, when they oviposit in rosettes. The long period of reproductive diapause constrains mass-rearing this weevil because only one generation per year can be produced. Determining the environmental conditions that regulate diapause termination may enable shortening diapause under laboratory conditions to increase production of adults to release. We tested three hibernating conditions (greenhouse [ambient temperature and photoperiod], glass door refrigerator [5°C and ambient photoperiod], and growth chamber [5°C and 24 h dark]) for three durations (4, 8, 11 wk). The highest proportion of females laying eggs came from the growth chamber, with 40% terminating diapause after 4 wk, 80% after 8 wk, and 95% after 11 wk of exposure. Our study demonstrates that duration of cold temperature is an important stimulus to terminate reproductive diapause of C. basicorne, and that exposure to ambient light had no effect at 5°C. However, 47% of females held at ambient greenhouse conditions, without any chilling period, completed diapause within 11 wk. Thus, a cold period can accelerate diapause development, but it is not necessary for its completion. Reducing the winter diapause period from about 6 mo to 11 wk should enable the production of multiple generations per year to increase the number of insects available to release.

Diapause differentially modulates the transcriptomes of fat body and flight muscle in the Colorado potato beetle

Many temperate insects, such as the Colorado potato beetle, enter diapause in winter, during which they arrest their development, suppress their metabolic rate and have high stress tolerance. Diapause phenotypes can be transcriptionally regulated, however many studies to date report only whole animal gene expression rather than tissue-specific processes during diapause. We used RNA-seq to measure gene expression in fat body and flight muscle of diapausing and non-diapausing beetles. We used differential expression and GO enrichment analyses to evaluate longstanding hypotheses about the mechanisms that drive arrested development, changes in energy metabolism, and increased stress tolerance during diapause. We found evidence of G2/M cell cycle arrest, juvenile hormone catabolism, increased antioxidant metabolism, epigenetic modification, transposable element regulation, and cytoskeletal remodeling in both the fat body and flight muscle of diapausing beetles. Beetles differentially modulated the fat body and flight muscle transcriptomes during diapause with fat body playing a larger role in the hypoxia response and immunity, whereas flight muscle had higher abundance of transcripts related to the chaperone response and proteostasis. Our transcriptome provides evidence for distinct roles and responses of fat body and flight muscle during diapause in the Colorado potato beetle, and we provide testable hypotheses for biological processes that appear to drive diapause phenotypes in insects.