Testing the evolutionary potential of an alpine plant: phenotypic plasticity in response to growth temperature outweighs parental environmental effects and other genetic causes of variation.

Phenotypic plasticity and rapid evolution are fundamental processes by which organisms can maintain their function and fitness in the face of environmental changes. Here we quantified the plasticity and evolutionary potential of an alpine herb Wahlenbergia ceracea. Utilizing its mixed-mating system, we generated outcrossed and self-pollinated families that were grown in either cool or warm environments, and that had parents that had also been grown in either cool or warm environments. We then analysed the contribution of environmental and genetic factors to variation in a range of phenotypic traits including phenology, leaf mass per area, photosynthetic function, thermal tolerance, and reproductive fitness. The strongest effect was that of current growth temperature, indicating strong phenotypic plasticity. All traits except thermal tolerance were plastic, whereby warm-grown plants flowered earlier, grew larger, and produced more reproductive stems compared with cool-grown plants. Flowering onset and biomass were heritable and under selection, with early flowering and larger plants having higher relative fitness. There was little evidence for transgenerational plasticity, maternal effects, or genotype×environment interactions. Inbreeding delayed flowering and reduced reproductive fitness and biomass. Overall, we found that W. ceracea has the capacity to respond rapidly to climate warming via plasticity, and the potential for evolutionary change.

Breeding phenology drives variation in reproductive output, reproductive costs, and offspring fitness in a viviparous ectotherm.

Phenological advances are a widespread response to global warming and can contribute to determine the climate vulnerability of organisms, particularly in ectothermic species, which are highly dependent on ambient temperatures to complete their life cycle. Yet, the relative contribution of breeding dates and temperature conditions during gestation on fitness of females and their offspring is poorly documented in reptiles. Here, we exposed females of the common lizard Zootoca vivipara to contrasting thermal scenarios (cold vs. hot treatment) during gestation and quantified effects of parturition dates and thermal treatment on life-history traits of females and their offspring for 1 year. Overall, our results suggest that parturition date has a greater impact than thermal conditions during gestation on life history strategies. In particular, we found positive effects of an earlier parturition date on juvenile survival, growth, and recruitment suggesting that environmental-dependent selection and/or differences in parental quality between early and late breeders underlie seasonal changes in offspring fitness. Yet, an earlier parturition date compromised the energetic condition of gravid females, which suggests the existence of a mother-offspring conflict regarding the optimization of parturition dates. While numerous studies focused on the direct effects of alterations in incubation temperatures on reptile life-history traits, our results highlight the importance of considering the role of breeding phenology in assessing the short- and long-term effects of thermal developmental plasticity.

Testing metabolic cold adaptation and the climatic variability hypothesis in two latitudinally distant populations of a supratidal water beetle.

Temperature significantly impacts ectotherm physiology, with thermal tolerance and metabolic traits typically varying with latitude across species ranges. The drivers of this variation remain unclear, however, despite obvious consequences for population persistence and conservation in the face of ongoing global change. This study explored local adaptation and phenotypic plasticity of metabolic rates and thermal limits in the supratidal rockpool beetle Ochthebius lejolisii. Using populations from localities at different ends of the species range that experience contrasting thermal variability, we simultaneously tested two of the major paradigms of spatial physiological ecology: metabolic cold adaptation (MCA) and the climatic variability hypothesis (CVH). Reciprocal acclimation was conducted under spring temperature regimes of both localities, incorporating local diurnal variation. Metabolic rates were measured by closed respirometry, and thermal tolerance limits estimated through thermography. In line with MCA, the higher-latitude population (colder climate) showed higher metabolic rates and temperature coefficients (Q10s) at lower temperatures than the lower-latitude population. As predicted by the CVH, the lower-latitude population (more variable climate) showed higher upper thermal tolerance but only the higher-latitude population was able to acclimate upper thermal limits. This result suggests trade-offs between physiological thermal limits and thermal plasticity in this species. A limited acclimation capacity could make populations on Mediterranean coasts especially vulnerable in the face of projected increases in extreme temperatures under ongoing climate change.

Phenotypic plasticity and thermal efficiency of temperature responses in two conspecific springtail populations from contrasting climates.

Temperature drives adaptation in life-history traits through direct effects on physiological processes. However, multiple life-history traits co-evolve as a life-history strategy. Therefore, physiological limitations constraining the evolution of trait means and phenotypic plasticity can be larger for some traits than the others. Comparisons of thermal responses across life-history traits can improve our understanding of the mechanisms determining the life-history strategies. In the present study, we focused on a soil microarthropod species abundant across the Northern Hemisphere, Folsomia quadrioculata (Collembola), with previously known effects of macroclimate. We selected an arctic and a temperate population from areas with highly contrasting climates - the arctic tundra and a coniferous forest floor, respectively - and compared them for thermal plasticity and thermal efficiency in growth, development, fecundity, and survival across four temperatures for a major part of their life cycle. We intended to understand the mechanisms by which temperature drives the evolution of life-history strategies. We found that the temperate population maximized performance at 10-15 °C, whereas the arctic population maintained its thermal efficiency across a wider temperature range (10-20 °C). Thermal plasticity varied in a trait-specific manner, and when considered together with differences in thermal efficiency, indicated that stochasticity in temperature conditions may be important in shaping the life-history strategies. Our study suggests that adopting a whole-organism approach and including physiological time considerations while analysing thermal adaptation will markedly improve our understanding of plausible links between thermal adaptation and responses to global climate change.

Thermal plasticity of wing size and wing spot size in Drosophila guttifera.

Thermal plasticity of melanin pigmentation patterns in Drosophila species has been studied as a model to investigate developmental mechanisms of phenotypic plasticity. The developmental process of melanin pigmentation patterns on wings of Drosophila is divided into two parts, prepattern specification during the pupal period and wing vein-dependent transportation of melanin precursors after eclosion. Which part can be affected by thermal changes? To address this question, we used polka-dotted melanin spots on wings of Drosophila guttifera, whose spot areas are specified by wingless morphogen. In this research, we reared D. guttifera at different temperatures to test whether wing spots show thermal plasticity. We found that wing size becomes larger at lower temperature and that different spots have different reaction norms. Furthermore, we changed the rearing temperature in the middle of the pupal period and found that the most sensitive developmental periods for wing size and spot size are different. The results suggest that the size control mechanisms for the thermal plasticity of wing size and spot size are independent. We also found that the most sensitive stage for spot size was part of the pupal period including stages at which wingless is expressed in the polka-dotted pattern. Therefore, it is suggested that temperature change might affect the prepattern specification process and might not affect transportation through wing veins.

Thermoregulatory trade-offs underlie the effects of warming summer temperatures on deer mice.

Climate warming could challenge the ability of endotherms to thermoregulate and maintain normal body temperature (Tb), but the effects of warming summer temperatures on activity and thermoregulatory physiology in many small mammals remain poorly understood. We examined this issue in deer mice (Peromyscus maniculatus), an active nocturnal species. Mice were exposed in the lab to simulated seasonal warming, in which an environmentally realistic diel cycle of ambient temperature (Ta) was gradually warmed from spring conditions to summer conditions (controls were maintained in spring conditions). Activity (voluntary wheel running) and Tb (implanted bio-loggers) were measured throughout, and indices of thermoregulatory physiology (thermoneutral zone, thermogenic capacity) were assessed after exposure. In control mice, activity was almost entirely restricted to the night-time, and Tb fluctuated ∼1.7°C between daytime lows and night-time highs. Activity, body mass and food consumption were reduced and water consumption was increased in later stages of summer warming. This was accompanied by strong Tb dysregulation that culminated in a complete reversal of the diel pattern of Tb variation, with Tb reaching extreme highs (∼40°C) during daytime heat but extreme lows (∼34°C) at cooler night-time temperatures. Summer warming was also associated with reduced ability to generate body heat, as reflected by decreased thermogenic capacity and decreased mass and uncoupling protein (UCP1) content of brown adipose tissue. Our findings suggest that thermoregulatory trade-offs associated with daytime heat exposure can affect Tb and activity at cooler night-time temperatures, impacting the ability of nocturnal mammals to perform behaviours important for fitness in the wild.

Sex-specific transgenerational plasticity: developmental temperatures of mothers and fathers have different effects on sons and daughters.

Each parent can influence offspring phenotype via provisioning of the zygote or sex-specific DNA methylation. Transgenerational plasticity may therefore depend on the environmental conditions experienced by each parent. We tested this hypothesis by conducting a fully factorial experiment across three generations of guppies (Poecilia reticulata), determining the effects of warm (28°C) and cold (21°C) thermal backgrounds of mothers and fathers on mass and length, and thermal performance (sustained and sprint swimming speeds, citrate synthase and lactate dehydrogenase activities; 18, 24, 28, 32 and 36°C test temperatures) of sons and daughters. Offspring sex was significant for all traits except for sprint speed. Warmer mothers produced sons and daughters with reduced mass and length, and warmer fathers produced shorter sons. Sustained swimming speed (Ucrit) of male offspring was greatest when both parents were raised at 28°C, and warmer fathers produced daughters with greater Ucrit. Similarly, warmer fathers produced sons and daughters with greater metabolic capacity. We show that the thermal variation experienced by parents can modify offspring phenotype, and that predicting the impacts of environmental change on populations would require knowledge of the thermal background of each mother and father, particularly where sexes are spatially segregated.

Increase in heat tolerance following a period of heat stress in a naturally occurring insect species.

Climate warming is the defining environmental crisis of the 21st century. Elucidating whether organisms can adapt to rapidly changing thermal environments is therefore a crucial research priority. We investigated warming effects on a native Hemipteran insect (Murgantia histrionica) that feeds on an endemic plant species (Isomeris arborea) of the California coastal sage scrub. Experiments conducted in 2009 quantified the temperature responses of juvenile maturation rates and stage-specific and cumulative survivorship. The intervening decade has seen some of the hottest years ever recorded, with increasing mean temperatures accompanied by an increase in the frequency of hot extremes. Experiments repeated in 2021 show a striking change in the bugs' temperature responses. In 2009, no eggs developed past the second nymphal stage at 33°C. In 2021, eggs developed into reproductive adults at 33°C. Upper thermal limits for maturation and survivorship have increased, along with a decrease in mortality risk with increasing age and temperature, and a decrease in the temperature sensitivity of mortality with increasing age. While we cannot exclude the possibility that other environmental factors occurring in concert could have affected our findings, the fact that all observed trait changes are in the direction of greater heat tolerance suggests that consistent exposure to extreme heat stress may at least be partially responsible for these changes. Harlequin bugs belong to the suborder Heteroptera, which contains a number of economically important pests, biological control agents and disease carriers. Their differential success in withstanding warming compared to beneficial holometabolous insects such as pollinators may exacerbate the decline of beneficial insects due to other causes (e.g. pollution and pesticides) with potentially serious consequences on both biodiversity and ecosystem functioning.

Rapid heat hardening in embryos of the lizard Anolis sagrei.

Adaptive thermal tolerance plasticity can dampen the negative effects of warming. However, our knowledge of tolerance plasticity is lacking for embryonic stages that are relatively immobile and may benefit the most from an adaptive plastic response. We tested for heat hardening capacity (a rapid increase in thermal tolerance that manifests in minutes to hours) in embryos of the lizard Anolis sagrei. We compared the survival of a lethal temperature exposure between embryos that either did (hardened) or did not (not hardened) receive a high but non-lethal temperature pre-treatment. We also measured heart rates (HRs) at common garden temperatures before and after heat exposures to assess metabolic consequences. 'Hardened' embryos had significantly greater survival after lethal heat exposure relative to 'not hardened' embryos. That said, heat pre-treatment led to a subsequent increase in embryo HR that did not occur in embryos that did not receive pre-treatment, indicative of an energetic cost of mounting the heat hardening response. Our results are not only consistent with adaptive thermal tolerance plasticity in these embryos (greater heat survival after heat exposure), but also highlight associated costs. Thermal tolerance plasticity may be an important mechanism by which embryos respond to warming that warrants greater consideration.

Thermal tolerance of the male freshwater prawn Cryphiops caementarius exposed to different acclimation temperatures.

The variation in water temperature influences metabolic and biochemical processes in ectothermic organisms, affecting development, behavior, and thermal responses. We conducted laboratory experiments based on different acclimation temperatures to determine the thermal tolerance in male specimens of the freshwater prawn Cryphiops caementarius. During 30 days, male prawns were exposed to acclimation temperatures of 19 °C (control), 24 °C, and 28 °C treatments. The Critical Thermal Maxima (CTMax) values at these acclimation temperatures were 33.42 °C, 34.92 °C, and 36.80 °C; whereas values for the Critical Thermal Minimum (CTMin) were 9.38 °C, 10.57 °C, and 13.88 °C. All acclimation temperature treatments had a positive effect (P < 0.05) on CTMax and CTMin, with high and significant correlations (CTMax: r = 0.992, P < 0.01; CTMin: r = 0.946, P < 0.01). The area of the thermal tolerance polygon over the three acclimation temperatures was 211.32 °C2 and the acclimation response rate values were high (CTMax from 0.30 to 0.47; CTMin from 0.24 to 0.83) but similar to those from other tropical crustacean species. These results demonstrate that adult males of the freshwater prawn C. caementarius can tolerate extreme water temperatures through a thermal plasticity response, which could be advantageous during a global warming scenario.

Windows of opportunity: Ocean warming shapes temperature-sensitive epigenetic reprogramming and gene expression across gametogenesis and embryogenesis in marine stickleback.

Rapid climate change is placing many marine species at risk of local extinction. Recent studies show that epigenetic mechanisms (e.g. DNA methylation, histone modifications) can facilitate both within and transgenerational plasticity to cope with changing environments. However, epigenetic reprogramming (erasure and re-establishment of epigenetic marks) during gamete and early embryo development may hinder transgenerational epigenetic inheritance. Most of our knowledge about reprogramming stems from mammals and model organisms, whereas the prevalence and extent of reprogramming among non-model species from wild populations is rarely investigated. Moreover, whether reprogramming dynamics are sensitive to changing environmental conditions is not well known, representing a key knowledge gap in the pursuit to identify mechanisms underlying links between parental exposure to changing climate patterns and environmentally adapted offspring phenotypes. Here, we investigated epigenetic reprogramming (DNA methylation/hydroxymethylation) and gene expression across gametogenesis and embryogenesis of marine stickleback (Gasterosteus aculeatus) under three ocean warming scenarios (ambient, +1.5 and +4°C). We found that parental acclimation to ocean warming led to dynamic and temperature-sensitive reprogramming throughout offspring development. Both global methylation/hydroxymethylation and expression of genes involved in epigenetic modifications were strongly and differentially affected by the increased warming scenarios. Comparing transcriptomic profiles from gonads, mature gametes and early embryonic stages showed sex-specific accumulation and temperature sensitivity of several epigenetic actors. DNA methyltransferase induction was primarily maternally inherited (suggesting maternal control of remethylation), whereas induction of several histone-modifying enzymes was shaped by both parents. Importantly, massive, temperature-specific changes to the epigenetic landscape occurred in blastula, a critical stage for successful embryo development, which could, thus, translate to substantial consequences for offspring phenotype resilience in warming environments. In summary, our study identified key stages during gamete and embryo development with temperature-sensitive reprogramming and epigenetic gene regulation, reflecting potential 'windows of opportunity' for adaptive epigenetic responses under future climate change.

Metabolic rate increases with acclimation temperature and is associated with mitochondrial function in some tissues of threespine stickleback.

The metabolic rate (MO2) of eurythermal fishes changes in response to temperature, yet it is unclear how changes in mitochondrial function contribute to changes in MO2. We hypothesized that MO2 would increase with acclimation temperature in the threespine stickleback (Gasterosteus aculeatus) in parallel with metabolic remodeling at the cellular level but that changes in metabolism in some tissues, such as liver, would contribute more to changes in MO2 than others. Threespine stickleback were acclimated to 5, 12 and 20°C for 7 to 21 weeks. At each temperature, standard and maximum metabolic rate (SMR and MMR, respectively), and absolute aerobic scope (AAS) were quantified, along with mitochondrial respiration rates in liver, oxidative skeletal and cardiac muscles, and the maximal activity of citrate synthase (CS) and lactate dehydrogenase (LDH) in liver, and oxidative and glycolytic skeletal muscles. SMR, MMR and AAS increased with acclimation temperature, along with rates of mitochondrial phosphorylating respiration in all tissues. Low SMR and MMR at 5°C were associated with low or undetectable rates of mitochondrial complex II activity and a greater reliance on complex I activity in liver, oxidative skeletal muscle and heart. SMR was positively correlated with cytochrome c oxidase (CCO) activity in liver and oxidative muscle, but not mitochondrial proton leak, whereas MMR was positively correlated with CCO activity in liver. Overall, the results suggest that changes in MO2 in response to temperature are driven by changes in some aspects of mitochondrial function in some, but not all, tissues of threespine stickleback.

Gene expression plasticity, genetic variation and fatty acid remodelling in divergent populations of a tropical bivalve species.

Ocean warming challenges marine organisms' resilience, especially for species experiencing temperatures close to their upper thermal limits. A potential increase in thermal tolerance might significantly reduce the risk of population decline, which is intrinsically linked to variability in local habitat temperatures. Our goal was to assess the plastic and genetic potential of response to elevated temperatures in a tropical bivalve model, Pinctada margaritifera. We benefit from two ecotypes for which local environmental conditions are characterized by either large diurnal variations in the tide pools (Marquesas archipelago) or low mean temperature with stable to moderate seasonal variations (Gambier archipelago). We explored the physiological basis of individual responses to elevated temperature, genetic divergence as well as plasticity and acclimation by combining lipidomic and transcriptomic approaches. We show that P. margaritifera has certain capacities to adjust to long-term elevated temperatures that was thus far largely underestimated. Genetic variation across populations overlaps with gene expression and involves the mitochondrial respiration machinery, a central physiological process that contributes to species thermal sensitivity and their distribution ranges. Our results present evidence for acclimation potential in P. margaritifera and urge for longer term studies to assess populations resilience in the face of climate change.

Le réchauffement des océans remet en question la résilience des organismes marins, en particulier pour les espèces connaissant des températures proches de leurs limites thermiques supérieures. Une augmentation potentielle de la tolérance thermique pourrait ainsi réduire considérablement le risque de déclin de la population. L'objectif de cette étude était d'évaluer le potentiel plastique et génétique de la réponse à l'exposition courte et chronique à températures élevées chez une espèce de bivalve tropical, Pinctada margaritifera. Ce modèle bénéficie de l'existence de deux écotypes pour lesquels les conditions environnementales locales sont caractérisées soit par de fortes variations diurnes associées aux marées (archipel des Marquises) soit par une température moyenne plus basse et des variations saisonnières prononcées (archipel des Gambier). Nous avons exploré les bases physiologiques des réponses individuelles ainsi que la divergence génétique et quantifié la plasticité en combinant des approches lipidomique et transcriptomique. Nous montrons que P. margaritifera possède des capacités d'acclimatation à des températures élevées sur le long terme jusqu'à présent largement sous-estimées. La divergence génétique entre populations est par ailleurs associée à des différences d'expression des gènes et implique la machinerie respiratoire mitochondriale, un processus physiologique central qui contribue à la sensibilité thermique des espèces et à leurs répartitions. Nos résultats présentent les bases des potentiels d'acclimatation chez P. margaritifera et soulignent l'importance d'études à plus long terme pour évaluer la résilience des populations face au changement climatique.

Temperature regime during embryogenesis alters subsequent behavioural phenotypes of juvenile brown trout.

Climate warming imposes a serious threat, especially to freshwater ecosystems in temperate and (sub)polar regions, which are often dominated by cold-adapted ectotherms. Although relatively intense warming during winter is common across the climatic regions, comparably little focus has been put on the organismal impacts of winter warming. Embryonic development, which is exceptionally susceptible to ambient temperature, occurs during winter in various freshwater ectotherms. Yet, our knowledge of the effects of increased temperature during embryogenesis on later life stages is limited. Using brown trout (Salmo trutta), we examined how a 1.5°C temperature increase from fertilization to hatching affects various traits at the onset of the free-swimming stage (i.e. a comparison between 3.5 and 5.0°C treatments). Although all hatchlings were kept at the same temperature (7.0°C) from hatching to the onset of the free-swimming stage for about two months, the temperature increase during embryogenesis substantially reduced key ecological behaviours, i.e. activity and exploration levels, at the onset of the free-swimming stage despite only marginal temperature effects on morphological and physiological traits at this stage. Given the importance of behavioural traits in early growth and survival, our study suggests a likely pathway through which subtle changes in mean winter temperature affect early fitness.

Membrane lipid metabolism, heat shock response and energy costs mediate the interaction between acclimatization and heat-hardening response in the razor clam Sinonovacula constricta.

Thermal plasticity on different time scales, including acclimation/acclimatization and heat-hardening response - a rapid adjustment for thermal tolerance after non-lethal thermal stress, can interact to improve the resilience of organisms to thermal stress. However, little is known about physiological mechanisms mediating this interaction. To investigate the underpinnings of heat-hardening responses after acclimatization in warm seasons, we measured thermal tolerance plasticity, and compared transcriptomic and metabolomic changes after heat hardening at 33 or 37°C followed by recovery of 3 or 24 h in an intertidal bivalve Sinonovacula constricta. Clams showed explicit heat-hardening responses after acclimatization in a warm season. The higher inducing temperature (37°C) caused less effective heat-hardening effects than the inducing temperature that was closer to the seasonal maximum temperature (33°C). Metabolomic analysis highlighted the elevated content of glycerophospholipids in all heat-hardened clams, which may help to maintain the structure and function of the membrane. Heat shock proteins (HSPs) tended to be upregulated after heat hardening at 37°C but not at 33°C, indicating that there was no complete dependency of heat-hardening effects on upregulated HSPs. Enhanced energy metabolism and decreased energy reserves were observed after heat hardening at 37°C, suggesting more energy costs during exposure to a higher inducing temperature, which may restrict heat-hardening effects. These results highlight the mediating role of membrane lipid metabolism, heat shock responses and energy costs in the interaction between heat-hardening response and seasonal acclimatization, and contribute to the mechanistic understanding of evolutionary change and thermal plasticity during global climate change.

Two reproductive traits show contrasting genetic architectures in Plantago lanceolata.

In many species, temperature-sensitive phenotypic plasticity (i.e., an individual's phenotypic response to temperature) displays a positive correlation with latitude, a pattern presumed to reflect local adaptation. This geographical pattern raises two general questions: (a) Do a few large-effect genes contribute to latitudinal variation in a trait? (b) Is the thermal plasticity of different traits regulated pleiotropically? To address the questions, we crossed individuals of Plantago lanceolata derived from northern and southern European populations. Individuals naturally exhibited high and low thermal plasticity in floral reflectance and flowering time. We grew parents and offspring in controlled cool- and warm-temperature environments, mimicking what plants would encounter in nature. We obtained genetic markers via genotype-by-sequencing, produced the first recombination map for this ecologically important nonmodel species, and performed quantitative trait locus (QTL) mapping of thermal plasticity and single-environment values for both traits. We identified a large-effect QTL that largely explained the reflectance plasticity differences between northern and southern populations. We identified multiple smaller-effect QTLs affecting aspects of flowering time, one of which affected flowering time plasticity. The results indicate that the genetic architecture of thermal plasticity in flowering is more complex than for reflectance. One flowering time QTL showed strong cytonuclear interactions under cool temperatures. Reflectance and flowering plasticity QTLs did not colocalize, suggesting little pleiotropic genetic control and freedom for independent trait evolution. Such genetic information about the architecture of plasticity is environmentally important because it informs us about the potential for plasticity to offset negative effects of climate change.

Beneficial developmental acclimation in reproductive performance under cold but not heat stress.

Thermal plasticity can help organisms coping with climate change. In this study, we analyse how laboratory populations of the ectotherm species Drosophila subobscura, originally from two distinct latitudes and evolving for several generations in a stable thermal environment (18 °C), respond plastically to new thermal challenges. We measured adult performance (fecundity traits as a fitness proxy) of the experimental populations when exposed to five thermal regimes, three with the same temperature during development and adulthood (15-15 °C, 18-18 °C, 25-25 °C), and two where flies developed at 18 °C and were exposed, during adulthood, to either 15 °C or 25 °C. Here, we test whether (1) flies undergo stress at the two more extreme temperatures; (2) development at a given temperature enhances adult performance at such temperature (i.e. acclimation), and (3) populations with different biogeographical history show plasticity differences. Our findings show (1) an optimal performance at 18 °C only if flies were subjected to the same temperature as juveniles and adults; (2) the occurrence of developmental acclimation at lower temperatures; (3) detrimental effects of higher developmental temperature on adult performance; and (4) a minor impact of historical background on thermal response. Our study indicates that thermal plasticity during development may have a limited role in helping adults cope with warmer - though not colder - temperatures, with a potential negative impact on population persistence under climate change. It also emphasizes the importance of analysing the impact of temperature on all stages of the life cycle to better characterize the thermal limits.

Cold acclimation triggers major transcriptional changes in Drosophila suzukii.

BACKGROUND: Insects have the capacity to adjust their physiological mechanisms during their lifetime to promote cold tolerance and cope with sublethal thermal conditions, a phenomenon referred to as thermal acclimation. The spotted wing drosophila, Drosophila suzukii, is an invasive fruit pest that, like many other species, enhances its thermotolerance in response to thermal acclimation. However, little is known about the underlying mechanisms of this plastic response. Here, we promoted flies' cold tolerance by gradually increasing acclimation duration (i.e. pre-exposure from 2 h to 9 days at 10 °C), and then compared transcriptomic responses of cold hardy versus cold susceptible phenotypes using RNA sequencing. RESULTS: Cold tolerance of D. suzukii increased with acclimation duration; the longer the acclimation, the higher the cold tolerance. Cold-tolerant flies that were acclimated for 9 days were selected for transcriptomic analyses. RNA sequencing revealed a total of 2908 differentially expressed genes: 1583 were up- and 1325 were downregulated in cold acclimated flies. Functional annotation revealed many enriched GO-terms among which ionic transport across membranes and signaling were highly represented in acclimated flies. Neuronal activity and carbohydrate metabolism were also enriched GO-terms in acclimated flies. Results also revealed many GO-terms related to oogenesis which were underrepresented in acclimated flies. CONCLUSIONS: Involvement of a large cluster of genes related to ion transport in cold acclimated flies suggests adjustments in the capacity to maintain ion and water homeostasis. These processes are key mechanisms underlying cold tolerance in insects. Down regulation of genes related to oogenesis in cold acclimated females likely reflects that females were conditioned at 10 °C, a temperature that prevents oogenesis. Overall, these results help to understand the molecular underpinnings of cold tolerance acquisition in D. suzukii. These data are of importance considering that the invasive success of D. suzukii in diverse climatic regions relates to its high thermal plasticity.

Cold acclimation triggers lipidomic and metabolic adjustments in the spotted wing drosophila Drosophila suzukii (Matsumara).

Chronic cold exposure is detrimental to chill susceptible insects that may accumulate chill injuries. To cope with deleterious effects of cold temperature, insects employ a variety of physiological strategies and metabolic adjustments, such as production of cryoprotectants, or remodeling of cellular membranes. Cold tolerance is a key element determining the fundamental niche of species. Because Drosophila suzukii is an invasive fruit pest, originating from East Asia, knowledge about its thermal biology is urgently needed. Physiological mechanisms underlying cold tolerance plasticity remain poorly understood in this species. Here, we explored metabolic and lipidomic modifications associated with the acquisition of cold tolerance in D. suzukii using Omics technologies (LC- and GC-MS/MS). In both cold-acclimated males and females, we observed physiological changes consistent with homeoviscous/homeophasic adaptation of membranes: reshuffling of phospholipid head groups and increasing unsaturation rate of fatty acids. Modification of fatty acids unsaturation were also observed in triacylglycerides, which would likely increase accessibility of lipid reserves. At the metabolic level, we observed clear-cut differentiation of metabolic profiles with cold-acclimated metabotypes showing accumulation of several potential cryoprotectants (sugars and amino acids). Metabolic pathway analyses indicated a remodeling of various processes, including purine metabolism and aminoacyl tRNA biosynthesis. These data provide a large-scale characterization of lipid rearrangements and metabolic pathway modifications in D. suzukii in response to cold acclimation and contribute to characterizing the strategies used by this species to modulate cold tolerance.

An impressive capacity for cold tolerance plasticity protects against ionoregulatory collapse in the disease vector Aedes aegypti.

The mosquito Aedes aegypti is largely confined to tropical and subtropical regions, but its range has recently been spreading to colder climates. As insect biogeography is tied to environmental temperature, understanding the limits of A. aegypti thermal tolerance and their capacity for phenotypic plasticity is important in predicting the spread of this species. In this study, we report on the chill coma onset (CCO) and recovery time (CCRT), as well as low-temperature survival phenotypes of larvae and adults of A. aegypti that developed or were acclimated to 15°C (cold) or 25°C (warm). Cold acclimation did not affect CCO temperatures of larvae but substantially reduced CCO in adults. Temperature and the duration of exposure both affected CCRT, and cold acclimation strongly mitigated these effects and increased rates of survival following prolonged chilling. Female adults were far less likely to take a blood meal when cold acclimated, and exposing females to blood (without feeding) attenuated some of the beneficial effects of cold acclimation on CCRT. Lastly, larvae suffered from haemolymph hyperkalaemia when chilled, but cold acclimation attenuated the imbalance. Our results demonstrate that A. aegypti larvae and adults have the capacity to acclimate to low temperatures, and do so at least in part by better maintaining ion balance in the cold. This ability for cold acclimation may facilitate the spread of this species to higher latitudes, particularly in an era of climate change.