No patterns in thermal plasticity along a latitudinal gradient in Drosophila simulans from eastern Australia

Phenotypic plasticity may be an important initial mechanism to counter environmental change, yet we know relatively little about the evolution of plasticity in nature. Species with widespread distributions are expected to have evolved higher levels of plasticity compared with those with more restricted, tropical distributions. At the intraspecific level, temperate populations are expected to have evolved higher levels of plasticity than their tropical counterparts. However, empirical support for these expectations is limited. In addition, no studies have comprehensively examined the evolution of thermal plasticity across life stages. Using populations of Drosophila simulans collected from a latitudinal cline spanning the entire east coast of Australia, we assessed thermal plasticity, measured as hardening capacity (the difference between basal and hardened thermal tolerance) for multiple measures of heat and cold tolerance across both adult and larval stages of development. This allowed us to explicitly ask whether the evolution of thermal plasticity is favoured in more variable, temperate environments. We found no relationship between thermal plasticity and latitude, providing little support for the hypothesis that temperate populations have evolved higher levels of thermal plasticity than their tropical counterparts. With the exception of adult heat survival, we also found no association between plasticity and ten climatic variables, indicating that the evolution of thermal plasticity is not easily predicted by the type of environment that a particular population occupies. We discuss these results in the context of the role of plasticity in a warming climate.     

Investigating thermal acclimation effects before and after a cold shock in Drosophila melanogaster using behavioural assays

Acclimation to environmental change can impose both costs and benefits to organisms. In this study we explored to what extent locomotor behaviour of Drosophila melanogaster is influenced by developmental temperature and adult temperature in both the laboratory and the field. Following development at 15, 25, or 31 °C, adult flies were tested for locomotor activity at all developmental temperatures in the laboratory before and after exposure to a cold shock and in the field for their ability to locate resources after a cold shock. Both test (15, 25, and 31 °C) and developmental temperatures strongly affected locomoter activity, with flies developed at 25 °C having the highest activity at all three test temperatures before the cold shock. After the cold shock flies developed at 15 °C had higher activity compared with flies developed at 25 and 31 °C when tested at 15 and 25 °C, and flies developed at 25 °C had the highest activity when tested at 31 °C. Furthermore, flies developed at 31 °C showed longer recovery times following the cold shock at test temperatures of 15 and 25 °C. However, flies acclimated at 15 °C during development did not recover faster at 15 and 25 °C compared with flies developed at 25 °C. There were no significant correlations between recovery time and locomotor activity at any of the test temperatures. Flies developed at 15 °C and exposed to a cold shock before release in the field were much more successful in locating a resource at low field temperatures compared with flies developed at 25 and 31 °C. Our results provide support for both the beneficial acclimation hypothesis and the optimal developmental temperature hypothesis, but the results are highly context dependent and change with the temperature experienced by the individual during its lifetime.     

Intraspecific variation in thermal acclimation and tolerance between populations of the winter ant,Prenolepis imparis

Thermal phenotypic plasticity, otherwise known as acclimation, plays an essential role in how organisms respond to short‐term temperature changes. Plasticity buffers the impact of harmful temperature changes; therefore, understanding variation in plasticity in natural populations is crucial for understanding how species will respond to the changing climate. However, very few studies have examined patterns of phenotypic plasticity among populations, especially among ant populations. Considering that this intraspecies variation can provide insight into adaptive variation in populations, the goal of this study was to quantify the short‐term acclimation ability and thermal tolerance of several populations of the winter ant, Prenolepis imparis. We tested for correlations between thermal plasticity and thermal tolerance, elevation, and body size. We characterized the thermal environment both above and below ground for several populations distributed across different elevations within California, USA. In addition, we measured the short‐term acclimation ability and thermal tolerance of those populations. To measure thermal tolerance, we used chill‐coma recovery time (CCRT) and knockdown time as indicators of cold and heat tolerance, respectively. Short‐term phenotypic plasticity was assessed by calculating acclimation capacity using CCRT and knockdown time after exposure to both high and low temperatures. We found that several populations displayed different chill‐coma recovery times and a few displayed different heat knockdown times, and that the acclimation capacities of cold and heat tolerance differed among most populations. The high‐elevation populations displayed increased tolerance to the cold (faster CCRT) and greater plasticity. For high‐temperature tolerance, we found heat tolerance was not associated with altitude; instead, greater tolerance to the heat was correlated with increased plasticity at higher temperatures. These current findings provide insight into thermal adaptation and factors that contribute to phenotypic diversity by revealing physiological variance among populations.     

Candidate genes and thermal phenotypes: identifying ecologically important genetic variation for thermotolerance in the Australian Drosophila melanogaster cline

Clinal variation in traits often reflects climatic adaptation; in Drosophila melanogaster clinal variation provides an opportunity to link variation in chromosomal inversions, microsatellite loci and various candidate genes to adaptive variation in traits. We undertook association studies with crosses from a single population of D. melanogaster from eastern Australia to investigate the association between genetic markers and traits showing clinal variation. By genotyping parents and phenotyping offspring, we minimized genotyping costs but had the power to detect association between markers and quantitative traits. Consistent with prior studies, we found strong associations between the clinal chromosomal inversion In(3R)Payne and markers within it, as well as among these markers. We also found an association between In(3L)Payne and one marker located within this inversion. Of the five predicted associations between markers and traits, four were detected (increased heat, decreased cold resistance and body size with the heat shock gene hsr-omega S, increased cold resistance with the inversion In(3L)Payne), while one was not detected (heat resistance and the heat shock gene hsp68). In a set of eight exploratory tests, we detected one positive association (between hsp23a and heat resistance) but no associations of heat resistance with alleles at the hsp26, hsp83, Desat 2, alpha-Gpdh, hsp70 loci, while cold resistance was not associated with Frost and Dca loci. These results confirm interactions between hsr-omega and thermal resistance, as well as between In(3L)Payne and cold resistance, but do not provide evidence for associations between thermal responses and alleles at other clinically varying marker genes.     

Basal cold but not heat tolerance constrains plasticity among Drosophila species (Diptera: Drosophilidae)

Thermal tolerance and its plasticity are important for understanding ectotherm responses to climate change. However, it is unclear whether plasticity is traded‐off at the expense of basal thermal tolerance and whether plasticity is subject to phylogenetic constraints. Here, we investigated associations between basal thermal tolerance and acute plasticity thereof in laboratory‐reared adult males of eighteen Drosophila species at low and high temperatures. We determined the high and low temperatures where 90% of flies are killed (ULT₉₀ and LLT₉₀, respectively) and also the magnitude of plasticity of acute thermal pretreatments (i.e. rapid cold‐ and heat‐hardening) using a standardized, species‐specific approach for the induction of hardening responses. Regression analyses of survival variation were conducted in ordinary and phylogenetically informed approaches. Low‐temperature pretreatments significantly improved LLT₉₀ in all species tested except for D. pseudoobscura, D. mojavensis and D. borealis. High‐temperature pretreatment only significantly increased ULT₉₀ in D. melanogaster, D. simulans, D. pseudoobscura and D. persimilis. LLT₉₀ was negatively correlated with low‐temperature plasticity even after phylogeny was accounted for. No correlations were found between ULT₉₀ and LLT₉₀ or between ULT₉₀ and rapid heat‐hardening (RHH) in ordinary regression approaches. However, after phylogenetic adjustment, there was a positive correlation between ULT₉₀ and RHH. These results suggest a trade‐off between basal low‐temperature tolerance and acute low‐temperature plasticity, but at high temperatures, increased basal tolerance was accompanied by increased plasticity. Furthermore, high‐ and low‐temperature tolerances and their plasticity are clearly decoupled. These results are of broad significance to understanding how organisms respond to changes in habitat temperature and the degree to which they can adjust thermal sensitivity.     

Rapid cold-hardening of Drosophila melanogaster in a field setting

Abstract.  Although laboratory studies demonstrate that cooling to ecologically relevant temperatures and/or at ecologically relevant rates induces rapid cold-hardening (RCH) in a variety of insects, little is known of the induction of RCH in nature. In the present study, caged Drosophila melanogaster (1–2 days posteclosion) from a colony established with flies collected locally are placed in a field setting (i.e. in approximately 4-cm deep leaf litter beneath an apple tree in Mount Pleasant, Michigan) during late afternoon (18.00 h EST; 05.00 h GMT). As the cage temperature falls from 22.1 ± 0.8 to 10.1 ± 0.1 °C between 18.00 and 06.00 h, the proportion of flies surviving a transfer to –6 °C for 1 h increases from 10.0 ± 6.2% to 68.1 ± 7.2%. When obtained from field cages, and then cooled from 23 °C at approximately 0.33 °C min⁻¹, more female flies remain behaviourally responsive (clinging to surfaces, exhibiting an active righting response, and/or climbing) at temperatures of 8–12 °C (24.00 h samples) or 7–12 °C (06.00 h samples), than do those sampled from cages kept in an incubator (23 °C). Field cooling reduces chill coma temperature from 8.7 ± 0.2 °C at 06.00 h to 7.1 ± 0.2 °C at 24.00 h, and to 6.6 ± 0.2 °C at 06.00 h. These data demonstrate that, in a recently collected culture of D. melanogaster , natural changes in microenvironmental temperature induce RCH that can benefit the organism at temperatures encountered in nature.     

Complexity of the cold acclimation response in Drosophila melanogaster

Insects can increase their resistance to cold stress when they are exposed to non-lethal conditions prior to the stress; these plastic responses are normally described only in terms of immediate effects on mortality. Here we examine in Drosophila melanogaster the short- and longer-term effects of different conditions on several measures of cold resistance, but particularly chill coma recovery. Short-term exposure to sublethal temperature (cold hardening) did not decrease chill coma recovery times even though it decreased mortality. Exposure to 12 degrees C for 2 days (acclimation) decreased chill coma recovery times for a range of stressful temperatures when flies were cultured at 25 degrees C, but did not usually affect recovery times when flies were cultured at 19 degrees C. In contrast, 2-day exposure to 12 degrees C decreased mortality regardless of rearing temperature. Rearing at 19 degrees C decreased mortality and chill coma recovery time relative to rearing at 25 degrees C. Acclimation increased the eclosion rate of eggs from stressed females, but did not affect development time or size of the offspring. These results indicate that plastic responses to cold in D. melanogaster are complex when resistance is scored in different ways, and that effects can extend across generations.     

Resistance to thermal stress in adult Drosophila buzzatii: acclimation and variation among populations

Survival after heat stress was examined in adult Drosophila buzzatii from seven localities. Males and females were conditioned by a non-lethal high temperature before exposure to a severe thermal stress. Variable times elapsed between conditioning, either as adults or pupae, and exposure to the stress were used, and experimental times to the stress ranged from 2 to 96 hours. Survival after stress varied among populations, and differences generally were consistent across conditioning treatments and across experiments. Resistance to thermal stress was much higher following one conditioning bout 2–4 hours before exposure to a severe stress than when the time elapsed between conditioning and exposure was increased to 24 or more hours. Significantly more adults survived the stress if conditioned 4 days before exposure to the thermal stress, either as adults or as pupae, than if not conditioned. The rank order of resistance roughly followed that predicted from the climatic conditions of the localities of origin.     

Beneficial acclimation and the Bogert effect

Few studies have examined the extent to which phenotypic plasticity in a given trait might be influenced by behavioural responses to an environmental cue. Regulatory behaviour might eliminate environmental variation such that little selection for physiological change would take place. Here, to test this Bogert effect on acclimation, we use two life-stages of a kelp fly that inhabit the same habitat, but differ profoundly in their behaviour. We predicted that when denied opportunities for behavioural regulation, mobile, though brachypterous adults would show a performance advantage in most thermal environments following acclimation to their preferred temperature(s). By contrast, in the less mobile larvae, that have a broader thermal preference, beneficial acclimation would be more evident. Ordered factor anova with orthogonal polynomial contrasts revealed that adults recovered faster from chill coma following any one of six short-term temperature treatments if they had been acclimated at low temperature, whilst larvae showed beneficial acclimation.     

Variation in genetic architecture of olfactory behaviour among wild-derived populations of Drosophila melanogaster

Odour-guided behaviour is a quantitative trait determined by many genes that are sensitive to gene-environment interactions. Different natural populations are likely to experience different selection pressures on the genetic underpinnings of chemosensory behaviour. However, few studies have reported comparisons of the quantitative genetic basis of olfactory behaviour in geographically distinct populations. We generated isofemale lines of Drosophila melanogaster from six populations in Argentina and measured larval and adult responses to benzaldehyde. There was significant variation within populations for both larval and adult olfactory behaviour and a significant genotype x sex interaction (GSI) for adult olfactory behaviour. However, there is substantial variation in the contribution of GSI to the total phenotypic variance among populations. Estimates of evolvability are orders of magnitude higher for larvae than for adults. Our results suggest that the potential for evolutionary adaptation to the chemosensory environment is greater at the larval feeding stage than at the adult reproductive stage.     

QTL for the thermotolerance effect of heat hardening, knockdown resistance to heat and chill-coma recovery in an intercontinental set of recombinant inbred lines of Drosophila melanogaster

The thermotolerance effect of heat hardening (also called short-term acclimation), knockdown resistance to high temperature (KRHT) with and without heat hardening and chill-coma recovery (CCR) are important phenotypes of thermal adaptation in insects and other organisms. Drosophila melanogaster from Denmark and Australia were previously selected for low and high KRHT, respectively. These flies were crossed to construct recombinant inbred lines (RIL). KRHT was higher in heat-hardened than in nonhardened RIL. We quantify the heat-hardening effect (HHE) as the ratio in KRHT between heat-hardened and nonhardened RIL. Composite interval mapping revealed a more complex genetic architecture for KRHT without heat-hardening than for KRHT in heat-hardened insects. Five quantitative trait loci (QTL) were found for KRHT, but only two of them were significant after heat hardening. KRHT and CCR showed trade-off associations for QTL both in the middle of chromosome 2 and the right arm of chromosome 3, which should be the result of either pleiotropy or linkage. The major QTL on chromosome 2 explained 18% and 27-33% of the phenotypic variance in CCR and KRHT in nonhardened flies, respectively, but its KRHT effects decreased by heat hardening. We discuss candidate loci for each QTL. One HHE-QTL was found in the region of small heat-shock protein genes. However, HHE-QTL explained only a small fraction of the phenotypic variance. Most heat-resistance QTL did not colocalize with CCR-QTL. Large-effect QTL for CCR and KRHT without hardening (basal thermotolerance) were consistent across continents, with apparent transgressive segregation for CCR. HHE (inducible thermotolerance) was not regulated by large-effect QTL.     

Insect thermal tolerance: what is the role of ontogeny,ageing and senescence?

Temperature has dramatic evolutionary fitness consequences and is therefore a major factor determining the geographic distribution and abundance of ectotherms. However, the role that age might have on insect thermal tolerance is often overlooked in studies of behaviour, ecology, physiology and evolutionary biology. Here, we review the evidence for ontogenetic and ageing effects on traits of high- and low-temperature tolerance in insects and show that these effects are typically pronounced for most taxa in which data are available. We therefore argue that basal thermal tolerance and acclimation responses (i.e. phenotypic plasticity) are strongly influenced by age and/or ontogeny and may confound studies of temperature responses if unaccounted for. We outline three alternative hypotheses which can be distinguished to propose why development affects thermal tolerance in insects. At present no studies have been undertaken to directly address these options. The implications of these age-related changes in thermal biology are discussed and, most significantly, suggest that the temperature tolerance of insects should be defined within the age-demographics of a particular population or species. Although we conclude that age is a source of variation that should be carefully controlled for in thermal biology, we also suggest that it can be used as a valuable tool for testing evolutionary theories of ageing and the cellular and genetic basis of thermal tolerance.     

Resistance to thermal stress in desert Drosophila

1. Four species of Drosophila are endemic to the Sonoran Desert of North America where daily and seasonal high temperatures exceed those experienced by other species in this genus. The close association between these species and their cactus hosts means that they reside only in the desert and raises the question as to whether they are better able to survive heat stress than are non-desert species of Drosophila . The tolerance of adult flies of the four desert species D. mojavensis, D. nigrospiracula, D. pachea and D. mettleri and the cosmopolitan D. simulans to acute heat stress was tested.
2. There was considerable variability among the desert endemic species with respect to survival following heat exposure. Two species, D. mojavensis and D. pachea , were more resistant at 44 °C and 46 °C than the others, with D. mettleri exhibiting similar heat stress resistance to D. simulans .
3. While there was no consistent influence of gender on heat resistance, younger flies (1-day-old) showed significantly greater survival than did older flies (7- or 14-days old).     

A comprehensive assessment of geographic variation in heat tolerance and hardening capacity in populations of Drosophila melanogaster from eastern Australia

We examined latitudinal variation in adult and larval heat tolerance in Drosophila melanogaster from eastern Australia. Adults were assessed using static and ramping assays. Basal and hardened static heat knockdown time showed significant linear clines; heat tolerance increased towards the tropics, particularly for hardened flies, suggesting that tropical populations have a greater hardening response. A similar pattern was evident for ramping heat knockdown time at 0.06 °C min^?1 increase. There was no cline for ramping heat knockdown temperature (CT_max) at 0.1 °C min^?1 increase. Acute (static) heat knockdown temperature increased towards temperate latitudes, probably reflecting a greater capacity of temperate flies to withstand sudden temperature increases during summer in temperate Australia. Larval viability showed a quadratic association with latitude under heat stress. Thus, patterns of heat resistance depend on assay methods. Genetic correlations in thermotolerance across life stages and evolutionary potential for critical thermal limits should be the focus of future studies.     

The effects of selection for cold tolerance on cross-tolerance to other environmental stressors in Drosophila melanogaster

Abstract  Cross tolerance, whereby tolerance to one environmental stress is correlated with tolerance to other stressors, is thought to be widespread in insects. We used lines of Drosophila melanogaster Meigen (Diptera: Drosophilidae) selected for survival at a 1-h exposure to −5°C to examine the extent to which this selection results in increased tolerance to other stresses, including high and low temperatures, desiccation and starvation. While selection improved tolerance to acute cold exposure and survival at −5°C, there was little effect of selection regime on tolerance to other stressors. There was no correlation between tolerances to any of the stressors, suggesting different mechanisms of tolerance. This supports arguments that correlations between stress tolerances during selection experiments with D. melanogaster may be coincidental. The magnitude of heat-hardening was apparently constrained by basal tolerance among lines, but the magnitude of the rapid cold-hardening response was not correlated with basal cold tolerance, implying that the relationship between inducible and basal tolerances differs at high and low temperatures.     

Correlated changes in thermotolerance traits and body color phenotypes in montane populations of Drosophila melanogaster: analysis of within- and between-population variations

Thermotolerance traits vary across geographical gradients but there is a lack of clinal variation in some Drosophila species. Thus, it is not clear whether thermotolerance or other correlated traits are the target of natural selection. In order to test selection responses, we investigated body melanization and thermotolerance traits in six altitudinal populations of Drosophila melanogaster . Based on rearing different geographical populations under uniform growth conditions at 21 °C (common garden experiments), clinal variations for cold resistance are in the direction opposite to heat resistance along an altitudinal gradient, that is darker flies from highland populations evidenced higher levels of cold resistance while lowland populations showed higher heat resistance. Phenotypic plastic responses for body melanization at 17–28 °C showed significant correlations with thermotolerance traits. At 17 °C, regression coefficients as a function of altitude are highly significant and positive for cold resistance but negative for heat knockdown. However, for flies reared at 28 °C, there is no elevational change in melanization as well as thermotolerance traits. Thus, both genetic and plastic changes of body melanization and thermotolerance traits suggest a correlated selection response. Further, within-population analyses of body melanization (based on dark, intermediate and light color phenotypes) showed significant associations with thermotolerance traits. Correlated variations in body melanization and thermal tolerances are associated with climatic thermal variability ( T _cv) but not with T _min. or T _max. along an altitudinal gradient.     

Temperature and photoperiod affect stress resistance traits in Drosophila melanogaster

The long‐term survival of species and populations depends on their ability to adjust phenotypic values to environmental conditions. In particular, the capability of dealing with environmental stress to buffer detrimental effects on fitness is considered to be of pivotal importance. Resistance traits are readily modulated by a wide range of environmental factors. In the present study, Drosophila melanogaster Meigen is used to investigate plastic responses to temperature and photoperiod in stress resistance traits. The results reveal that stress resistance traits (cold, heat, starvation and desiccation resistance) are affected by the factors temperature and sex predominantly. Cooler temperatures compared with warmer temperatures increase cold tolerance, desiccation and starvation resistance, whereas they reduce heat tolerance. Except for heat resistance, females are more stress‐resistant than males. Stress resistance traits are also affected by photoperiod. Shorter photoperiods decrease cold tolerance, whereas longer photoperiods enhance desiccation resistance. Overall, thermal effects are pervasive throughout all measured resistance traits, whereas photoperiodic effects are of limited importance in the directly developing (i.e. nondiapausing) flies used here, suggesting that pronounced photoperiodic effects on stress resistance traits may be largely limited to, and triggered by, diapause‐inducing effects.