A trade-off between desiccation resistance and developmental humidity for pupation height in the North Indian seasonal population of Drosophilid-Zaprionus indianus.

Drosophila larvae and pupae are vulnerable to seasonal abiotic stressors such as humidity and temperature. In wild low-humidity habitats, desiccation stress can occur as Drosophila larvae forsake wet food in search of a drier pupation site. Henceforth, the hypothesis that developmental humidity impacts pupation height, affecting larval and pupae water balance and fitness-related traits, was examined. Accordingly, warm-adapted Drosophilid- Zaprionus indianus from two seasons were reared under season-specific simulated conditions, with significantly varying relative humidity (summer RH: 40%; rainy RH: 80%), but nearly identical temperatures. A trade-off between pupation height and developmental humidity was observed. Drier summer conditions lead to pupae wandering farther from drier glass surfaces, resulting in higher pupation height (17.3 cm) while rainy pupae prefer wet food, resulting in lower pupation height (7.12 cm). Additionally, density-dependent pupation height was developmental humidity-specific, with most rainy-season pupae pupated on wetter food, while dry summer pupae pupated on glass surfaces or cotton. Nevertheless, flies from far pupation exhibited greater desiccation resistance, fecundity, and copulation duration than those from near pupation. The cuticular lipid mass of larvae and pupae was higher during far-than-near pupation, indicating decreased water loss rates compared to near-pupation. Finally, pupae eclosion (%) was unaffected by greater humidity (85%) in either season. Still, it considerably decreased at lower humidity (RH: 0% and 38%) for rainy pupae, further supporting the selection of low-humidity desiccation resistance in pupae. In conclusion, low humidity is crucial for survival of pre-adult stages of Zaprionus indianus under desiccation stress and for preference of pupation site.

Adaptive changes in energy reserves and effects of body melanization on thermal tolerance in Drosophila simulans.

Seasonally polyphenic types have been documented in many Drosophilids, which differ significantly during thermal stress. Although Drosophila simulans is a sibling species to Drosophila melanogaster, both thrive in the temperate and tropical climates, but various climatic factors are expected to impact their distribution and abundance. As a result, D. simulans may use phenotypic plasticity to adapt to colder and drier circumstances in temperate zones, although such studies are less known. In the present study, our main aim was to find a link between adaptive plasticity and thermal tolerance in D. simulans. We characterized two morphs in D. simulans flies based on the abdominal melanization collected from the same locality and season, as this trait is highly associated with the larval developmental conditions. Our results suggested that flies reared from dark and light morph showed significant differences in the basal level of proline, carbohydrates (trehalose, glycogen), and lipids (cuticular lipids and total body lipids) within simulated seasons and morph lineages in D. simulans flies. We further showed that D. simulans reared from dark morph are better adapted to cold conditions, whereas light flies are more adapted to warm conditions. The flies, both from light and dark morph lineages, when reared at 15 °C, showed an increase in the level of total body lipids after acclimation at 0 °C but a decrease in the level of proline and carbohydrates (trehalose, glycogen). Heat acclimation increases glycogen levels in the flies from light morph lineage while decreases trehalose and proline.

Developmental and adult acclimation impact cold and drought survival of invasive tropical Drosophila kikkawai.

Narrow distribution patterns of tropical Drosophila species are limited by lower resistance to cold or drought. In the invasive tropical Drosophila kikkawai, we tested whether developmental and adult acclimations at cooler temperatures could enhance its stress resistance level. Adult acclimation of winter collected body color morphs revealed a significant increase in the level of cold resistance. For light morph, its abundance during winter is not consistent with thermal-melanism hypothesis. However, higher cold acclimation capacity, as well as storage of energy metabolites could support its winter survival. In the wild-caught light and intermediate morphs, there is a lack of trade-off between cold and heat resistance but not in the case of dark morph. Developmental plasticity (15°C) resulted in the fivefold increase of cold survival at 0°C; and a twofold increase in desiccation resistance but a modest reduction (∼28-35%) in heat resistance as compared to morph strains reared at 25°C. Drought acclimation changes were significantly higher as compared with cold or heat pretreatment. We observed a trade-off between basal resistance and acclimation capacity for cold, heat, or drought resistance. For homeostatic energy balance, adult acclimation responses (cold versus drought; heat versus drought) caused compensatory plastic changes in the levels of proline or trehalose (shared patterns) but different patterns for total body lipids. In contrast, rapid cold or heat hardening-induced changes in energy metabolites were different as compared to acclimation. The ability of D. kikkawai to significantly increase stress tolerance through plasticity is likely to support its invasion potential.

Plastic changes in cold and drought tolerance of Drosophila nepalensis correlate with sex-specific differences in body melanization, cuticular lipid mass, proline accumulation, and seasonal abundance.

Autumn-collected flies of Himalayan Drosophila nepalensis differ in body color phenotypes (males more melanized relative to females) and in their behavior (males abundant in the open sites vs. shelters-seeking females). In contrast, winter-collected flies of both sexes are equally melanized and abundant in the open sites. We tested developmental and adult plasticity changes in cold or drought tolerance in D. nepalensis flies reared under winter or autumn simulated conditions. In D. nepalensis flies reared at 21 °C, male flies were more cold tolerant (as shown by shorter chill-coma recovery time and lower cold-shock mortality). Further, male flies also exhibited greater drought tolerance (increased levels of desiccation resistance, cuticular lipid mass, melanization, hydration level, and dehydration tolerance) as compared to females. We observed sex-specific differences in the adult plasticity responses due to rapid cold or drought hardening (RCH or RDH); and for the persistence of cold acclimation effects. RCH or RDH-induced changes in the level of proline accumulations are negatively correlated with a decrease in the chill-coma recovery time. Therefore, cold or drought hardening treatments are likely to influence cold tolerance through proline accumulation. Developmental acclimation and adult hardening responses revealed significant interaction effects between sexes and thermal treatments. Thus, sex-specific differences in morphological traits (body melanization and cuticular lipid mass) and physiological traits (adult plasticity changes in cold tolerance and proline accumulation) correlate with behavioral divergence (habitat usage) across sexes.

Cold acclimation conditions constrain plastic responses for resistance to cold and starvation in Drosophila immigrans.

In montane Drosophila species, cold-induced plastic changes in energy metabolites are likely developed to cope with cold and starvation stress. Adult Drosophila immigrans reared at 15°C were acclimated at 0°C or 7°C for durations of up to 6 days (fed or unfed conditions). Such flies were tested for plastic changes in resistance to cold or starvation stress as well as for possible accumulation and utilization of four energy metabolites (body lipids, proline, trehalose and glycogen). Adults acclimated at 7°C revealed a greater increase in cold tolerance than flies acclimated at 0°C. Different durations of cold acclimation at 7°C led to increased level of body lipids only in fed flies which were utilized under starvation stress. However, such plastic responses were not observed in the flies acclimated at 0°C, which remained unfed due to chill-coma. These observations suggest a possible role of feeding to improve starvation resistance only in the flies acclimated at 7°C with food. Cold acclimated D. immigrans flies revealed improved cold resistance through a possible reshuffling of trehalose and glycogen; and starvation-induced proline which was utilized under cold stress durations. Finally, greater reduction in mean daily fecundity due to cold or starvation was observed in 0°C acclimated flies as compared to 7°C acclimated flies. Thus, cold acclimation conditions (0°C or 7°C) greatly impact resistance to cold and starvation in D. immigrans.

Heat- and humidity-induced plastic changes in body lipids and starvation resistance in the tropical fly Zaprionus indianus during wet and dry seasons.

Insects in tropical wet or dry seasons are likely to cope with starvation stress through plastic changes (developmental as well as adult acclimation) in energy metabolites. Control and experimental groups of Zaprionus indianus flies were reared under wet or dry conditions, but adults were acclimated at different thermal or humidity conditions. Adult flies of the control group were acclimated at 27°C and low (50%) or high (60%) relative humidity (RH). For experimental groups, adult flies were acclimated at 32°C for 1 to 6 days and under low (40%) or high (70%) RH. For humidity acclimation, adult flies were acclimated at 27°C but under low (40%) or high (70%) RH for 1 to 6 days. Plastic changes in experimental groups as compared with the control group (developmental as well as adult acclimation) revealed significant accumulation of body lipids owing to thermal or humidity acclimation of wet season flies, but low humidity acclimation did not change the level of body lipids in dry season flies. Starvation resistance and body lipids were higher in the males of dry season flies but in the females of wet season flies. Adults acclimated under different thermal or humidity conditions exhibited changes in the rate of utilization of body lipids, carbohydrates and proteins. Adult acclimation of wet or dry season flies revealed plastic changes in mean daily fecundity; and a reduction in fecundity under starvation. Thus, thermal or humidity acclimation of adults revealed plastic changes in energy metabolites to support starvation resistance of wet or dry season flies.

Tropical Drosophila ananassae of wet-dry seasons show cross resistance to heat, drought and starvation.

Plastic responses to multiple environmental stressors in wet or dry seasonal populations of tropical Drosophila species have received less attention. We tested plastic effects of heat hardening, acclimation to drought or starvation, and changes in trehalose, proline and body lipids in Drosophila ananassae flies reared under wet or dry season-specific conditions. Wet season flies revealed significant increase in heat knockdown, starvation resistance and body lipids after heat hardening. However, accumulation of proline was observed only after desiccation acclimation of dry season flies while wet season flies elicited no proline but trehalose only. Therefore, drought-induced proline can be a marker metabolite for dry-season flies. Further, partial utilization of proline and trehalose under heat hardening reflects their possible thermoprotective effects. Heat hardening elicited cross-protection to starvation stress. Stressor-specific accumulation or utilization as well as rates of metabolic change for each energy metabolite were significantly higher in wet-season flies than dry-season flies. Energy metabolite changes due to inter-related stressors (heat versus desiccation or starvation) resulted in possible maintenance of energetic homeostasis in wet- or dry-season flies. Thus, low or high humidity-induced plastic changes in energy metabolites can provide cross-protection to seasonally varying climatic stressors.

Cross-tolerance effects due to adult heat hardening, desiccation and starvation acclimation of tropical drosophilid-Zaprionus indianus.

Some insect taxa from polar or temperate habitats have shown cross-tolerance for multiple stressors but tropical insect taxa have received less attention. Accordingly, we considered adult flies of a tropical drosophilid-Zaprionus indianus for testing direct as well as cross-tolerance effects of rapid heat hardening (HH), desiccation acclimation (DA) and starvation acclimation (SA) after rearing under warmer and drier season specific simulated conditions. We observed significant direct acclimation effects of HH, DA and SA; and four cases of cross-tolerance effects but no cross-tolerance between desiccation and starvation. Cross-tolerance due to heat hardening on desiccation showed 20% higher effect than its reciprocal effect. There is greater reduction of water loss in heat hardened flies (due to increase in amount of cuticular lipids) as compared with desiccation acclimated flies. However, cross-tolerance effect of SA on heat knockdown was two times higher than its reciprocal. Heat hardened and desiccation acclimated adult flies showed substantial increase in the level of trehalose and proline while body lipids increased due to heat hardening or starvation acclimation. However, maximum increase in energy metabolites was stressor specific i.e. trehalose due to DA; proline due to HH and total body lipids due to SA. Rapid changes in energy metabolites due to heat hardening seem compensatory for possible depletion of trehalose and proline due to desiccation stress; and body lipids due to starvation stress. Thus, observed cross-tolerance effects in Z. indianus represent physiological changes to cope with multiple stressors related to warmer and drier subtropical habitats.

Cold and desiccation stress induced changes in the accumulation and utilization of proline and trehalose in seasonal populations of Drosophila immigrans.

Changes in the levels of energy metabolites can limit survival ability of Drosophila species under stressful conditions but this aspect has received less attention in wild populations collected in different seasons. We tested cold or desiccation triggered changes in the accumulation or utilization of two energy metabolites (trehalose and proline) in Drosophila immigrans flies reared under season specific environmental conditions. Such D.immigrans populations were subjected to different durations of cold (0°C) or desiccation stress (5% RH) or dual stress. We found stress induced effects of cold vs desiccation on the levels of trehalose as well as for proline. Different durations of cold stress led to accumulation of trehalose while desiccation stress durations revealed utilization of trehalose. In contrast, there was accumulation of proline under desiccation and utilization of proline with cold stress. Since accumulation levels were higher than utilization of each energy metabolite, the effects of dual stress showed additive effect. However, there was no utilization of total body lipids under cold or desiccation stress. We observed significant season specific differences in the amount of energy metabolites but the rate of metabolism did not vary across seasons. Stress triggered changes in trehalose and proline suggest possible link between desiccation and cold tolerance. Finally, stress specific (cold or desiccation) compensatory changes in the levels of trehalose and proline suggest possible energetic homeostasis in D.immigrans living under harsh climatic conditions of montane localities.

Effects of saturation deficit on desiccation resistance and water balance in seasonal populations of the tropical drosophilid Zaprionus indianus.

Seasonally varying populations of ectothermic insect taxa from a given locality are expected to cope with simultaneous changes in temperature and humidity through phenotypic plasticity. Accordingly, we investigated the effect of saturation deficit on resistance to desiccation in wild-caught flies from four seasons (spring, summer, rainy and autumn) and corresponding flies reared in the laboratory under season-specific simulated temperature and humidity growth conditions. Flies raised under summer conditions showed approximately three times higher desiccation resistance and increased levels of cuticular lipids compared with flies raised in rainy season conditions. In contrast, intermediate trends were observed for water balance-related traits in flies reared under spring or autumn conditions but trait values overlapped across these two seasons. Furthermore, a threefold difference in saturation deficit (an index of evaporative water loss due to a combined thermal and humidity effect) between summer (27.5 mB) and rainy (8.5 mB) seasons was associated with twofold differences in the rate of water loss. Higher dehydration stress due to a high saturation deficit in summer is compensated by storage of higher levels of energy metabolite (trehalose) and cuticular lipids, and these traits correlated positively with desiccation resistance. In Z. indianus, the observed changes in desiccation-related traits due to plastic effects of simulated growth conditions correspond to similar changes exhibited by seasonal wild-caught flies. Our results show that developmental plastic effects under ecologically relevant thermal and humidity conditions can explain seasonal adaptations for water balance-related traits in Z. indianus and are likely to be associated with its invasive potential.

Sex-specific divergence for body size and desiccation-related traits in Drosophila hydei from the western Himalayas.

Sex-specific-differences are a widespread source of genetic variation in various Drosophila species. In the present study, we have examined desiccation survival in males and females of Drosophila hydei from colder and drier montane conditions of the western Himalayas (altitudinal populations; 600-2202 m). In contrast with most other studies in drosophilids, D. hydei males exhibited comparatively higher desiccation resistance despite smaller body size compared to females. Accordingly, we tested the physiological basis of such adaptations in both sexes of D. hydei. Body size traits (wing length, wet weight and dry weight) were ~1.2 fold higher in females than males. However, desiccation resistance was 10 to 13 h higher in males than females. These differences matched enhanced storage of trehalose content (~1.2 fold), higher hemolymph content (~1.2 fold) and enhanced cuticular lipid mass (~1.5 fold) in males than females. Water loss before succumbing to death (dehydration tolerance) was much higher in males (~81%) than females (~64%). A greater loss of hemolymph water until death under desiccation stress was associated with higher desiccation resistance in males. Further, there were lacks of differences in the rate of water loss, rate of trehalose utilization and rate of hemolymph depletion between the sexes in D. hydei. Therefore, sex-specific differences in desiccation resistance of D. hydei were independent of body size as well as the exhaustion of metabolite reserves and rather were caused by the higher dehydration tolerance as well as higher acquisition of hemolymph and trehalose contents.

Developmental acclimation to low or high humidity conditions affect starvation and heat resistance of Drosophila melanogaster.

Several Drosophila species originating from tropical humid localities are more resistant to starvation and heat stress than populations from high latitudes but mechanistic bases of such physiological changes are largely unknown. In order to test whether humidity levels affect starvation and heat resistance, we investigated developmental acclimation effects of low to high humidity conditions on the storage and utilization of energy resources, body mass, starvation survival, heat knockdown and heat survival of D. melanogaster. Isofemale lines reared under higher humidity (85% RH) stored significantly higher level of lipids and showed greater starvation survival hours but smaller in body size. In contrast, lines reared at low humidity evidenced reduced levels of body lipids and starvation resistance. Starvation resistance and lipid storage level were higher in females than males. However, the rate of utilization of lipids under starvation stress was lower for lines reared under higher humidity. Adult flies of lines reared at 65% RH and acclimated under high or low humidity condition for 200 hours also showed changes in resistance to starvation and heat but such effects were significantly lower as compared with developmental acclimation. Isofemale lines reared under higher humidity showed greater heat knockdown time and heat-shock survival. These laboratory observations on developmental and adult acclimation effects of low versus high humidity conditions have helped in explaining seasonal changes in resistance to starvation and heat of the wild-caught flies of D. melanogaster. Thus, we may suggest that wet versus drier conditions significantly affect starvation and heat resistance of D. melanogaster.

Effects of body-size variation on flight-related traits in latitudinal populations of Drosophila melanogaster.

In the present study, we tested the hypothesis whether flight-related traits such as wing area, flight-muscle ratio, wing loading and dispersal yield evidence of geographical variation in nine wild-collected as well as laboratory-reared (at 21°C) latitudinal populations of Drosophila melanogaster from the Indian subcontinent. We observed positive clinal variation in the wing-thorax ratio, wing aspect ratio and wing area, along a latitudinal gradient for both the sexes. In contrast, geographical changes in three parameters of flight ability, i.e. flight-muscle ratio, wing loading and dispersal, showed negative correlation with latitude. On the basis of isofemale line variability, we observed positive correlation of wing loading with flight-muscle ratio as well as dispersal behaviour in both the sexes. We also found positive correlation between duration of development and wing area. Interestingly, southern populations of D. melanogaster from warm and humid habitats exhibited higher flight-muscle ratio as well as the higher wing loading than northern populations which occur in cooler and drier climatic conditions. Laboratory tests for dispersal-related walking behaviour showed significantly higher values for southern populations compared with northern populations of D. melanogaster. Multiple regression analysis of geographical changes in flight-muscle ratio, wing loading as well as walking behaviour as a function of average temperature and relative humidity of the origin of populations in wild-collected flies have suggested adaptive changes in flight-related traits in response to steeper gradients of climatic factors in the Indian subcontinent. Finally, adaptive latitudinal variations in flight-related traits in D. melanogaster are consistent with results of other studies from different continents despite differences due to specific climatic conditions in the Indian subcontinent.

Divergent strategies for adaptations to stress resistance in two tropical Drosophila species: effects of developmental acclimation in D. bipectinata and the invasive species D. malerkotliana.

Previous studies on two tropical Drosophila species (D. malerkotliana and D. bipectinata) have shown lower resistance to stress-related traits but the rapid colonization of D. malerkotliana in the past few decades is not consistent with its sensitivity to desiccation and cold stress. We tested the hypothesis that developmental acclimation at two growth temperatures (17 and 25°C) can confer adaptations to desiccation and thermal stresses. We found divergence in developmental plastic effects on cuticular traits, i.e. a significant increase of body melanisation (~2-fold) and of cuticular lipid mass (~3-fold) in D. malerkotliana but only 1.5-fold higher cuticular lipid mass in D. bipectinata when grown at 17°C compared with 25°C. A comparison of the water budget of these two species showed significantly higher effects of developmental acclimation on body water content, rate of water loss and dehydration tolerance resulting in higher desiccation resistance in D. malerkotliana than in D. bipectinata. When grown in cooler conditions (17°C), D. malerkotliana had greater resistance to cold as well as desiccation stress. In contrast, heat resistance of D. bipectinata was higher when grown at 25°C. These laboratory observations are supported by data on seasonally varying populations. Furthermore, adult D. malerkotliana acclimated to different stresses showed greater resistance to those stresses than D. bipectinata adults. Thus, significant increase in stress resistance of D. malerkotliana through developmental acclimation may be responsible for its invasion and ecological success on different continents compared with D. bipectinata.

Divergence of water balance mechanisms and acclimation potential in body color morphs of Drosophila ananassae.

Drosophila ananassae is a desiccation sensitive species, but the physiological basis of its abundance in the drier subtropical areas is largely unknown. We tested the hypothesis whether body color morphs of D. ananassae differ in the mechanistic basis of water conservation as well as desiccation acclimation potential, consistent with their distribution under dry or wet habitats. We observed reduced rate of water loss consistent with the greater desiccation potential of dark morph as compared with light morph, despite lack of quantitative differences in cuticular lipid mass between them. Dark morph evidenced greater wet and dry mass (∼1.17-fold) as well as higher hemolymph content (∼1.70-fold) and (∼17%) dehydration tolerance to sustain longer survival under desiccation stress (LT50 17.5 hr) as compared with light morph (LT50 4.3 hr). We found significant differences in the storage of energy metabolites in the body color morphs of D. ananassae, that is, carbohydrate content was significantly higher (∼0.18 mg/mg dry mass) in the dark morph as compared to light morph, but greater (∼0.05 mg/mg dry mass) body lipid content was evident in the light morph. Under desiccation stress, dark and light morphs utilized mainly carbohydrates but also lipids to a lesser extent. However, the rate of utilization of energy metabolites did not vary between dark and light morphs. Further, the dark morph consumed higher energy content derived from carbohydrates under desiccation stress as compared with the light morph. Finally, we found contrasting patterns of acclimation to desiccation stress in the two body color morphs, that is, increase in desiccation survival (4.7 hr), as well as in dehydration tolerance (∼6%) due to acclimation of the dark morph but no such effects were observed in the light morph. Thus, divergence in water balance mechanisms as well as acclimation potential reflects evolved physiological adaptations of the dark morph under drier but of the light morph to wet climatic conditions.

Sex-specific differences in desiccation resistance and the use of energy metabolites as osmolytes in Drosophila melanogaster flies acclimated to dehydration stress.

In the Indian subcontinent, there are significant between-population variations in desiccation resistance in Drosophila melanogaster, but the physiological basis of adult acclimation responses to ecologically relevant humidity conditions is largely unknown. We tested the hypothesis that increased desiccation resistance in acclimated flies is associated with changes in cuticular permeability and/or content of energy metabolites that act as osmolytes. Under an ecologically relevant humidity regime (~50 % relative humidity), both sexes showed desiccation acclimation which persisted for 2-3 days. However, only females responded to acclimation at ~5 % relative humidity (RH). Acclimated flies exhibited no changes in the rate of water loss, which is consistent with a lack of plastic changes in cuticular traits (body melanization, epicuticular lipid). Therefore, changes in cuticular permeability are unlikely in drought-acclimated adult flies of D. melanogaster. In acclimated flies, we found sex differences in changes in the content of osmolytes (trehalose in females versus glycogen in males). These sex-specific changes in osmolytes are rapid and reversible and match to corresponding changes in the increased desiccation resistance levels of acclimated flies. Further, the increased content of trehalose in females and glycogen in males support the bound-water hypothesis for water retention in acclimated flies. Thus, drought acclimation in adult flies of D. melanogaster involves inducible changes in osmolytes (trehalose and glycogen), while there is little support for changes in cuticular permeability.

Seasonal changes in humidity impact drought resistance in tropical Drosophila leontia: testing developmental effects of thermal versus humidity changes.

Drosophila leontia is native to highly humid equatorial tropical habitats but its desiccation sensitivity (~10h) is not consistent with its abundance during the drier autumn season in the subtropical regions. We have tested the effects of developmental acclimation on desiccation resistance and water balance related traits of D. leontia collected during rainy and autumn seasons. The isofemale lines of seasonal populations were reared under ecologically relevant growth temperatures (18 or 26 °C) or humidity conditions (35 or 85% RH) but tested at different times under identical experimental conditions. The larvae as well as flies reared under two thermal conditions (18 or 26 °C) showed no effect on desiccation related traits as well as storage and utilization of energy metabolites. In contrast, for D. leontia reared under low humidity (35% RH), significant changes at larval as well adult stages include increase in the desiccation resistance as well as cuticular lipid quantity, reduced levels of rate of body water loss, higher storage of carbohydrates but lower rate of utilization of carbohydrates as compared with flies reared at high humidity (85% RH). D. leontia has responded to rearing under low humidity conditions by increasing its desiccation resistance but not due to changes in the growth temperatures. These laboratory observations on seasonal populations highlight differences due to rearing conditions but not due to seasons. Further, direct analysis of wild-caught seasonal populations has shown trends similar to developmental acclimation effects. For wild caught flies, there are significant seasonal differences i.e. higher desiccation resistance as well as cuticular lipid quantity but reduced rate of water loss for autumn than rainy season flies. Thus, our laboratory observations are relevant for understanding seasonal adaptations of natural populations of tropical D. leontia to wet-dry conditions in the wild.

Sex-specific divergence for adaptations to dehydration stress in Drosophila kikkawai.

Several studies on diverse Drosophila species have reported higher desiccation resistance of females, but the physiological basis of such sex-specific differences has received less attention. We tested whether sex-specific differences in cuticular traits (melanic females and non-melanic males) of Drosophila kikkawai correspond with divergence in their water balance mechanisms. Our results are interesting in several respects. First, positive clinal variation in desiccation resistance was correlated with cuticular melanisation in females but with changes in cuticular lipid mass in males, despite a lack of differences between the sexes for the rate of water loss. Second, a comparative analysis of water budget showed that females of the northern population stored more body water as well as hemolymph content and exhibited greater dehydration tolerance than flies from the southern tropics. In contrast, we found no geographical variation in the males for water content and dehydration tolerance. Third, an ~10-fold increase in the rate of water loss after organic solvent treatment of male D. kikkawai suggested a role of cuticular lipids in cuticular transpiration, but had no effect in the females. Fourth, geographical differences in the storage of carbohydrate content (metabolic fuel) were observed in females but not in males. Interestingly, in females, the rate of utilization of carbohydrates did not vary geographically, but males from drier localities showed a 50% reduction compared with wetter localities. Thus, body melanisation, increased body water, hemolymph, carbohydrate content and greater dehydration tolerance confer greater desiccation resistance in females, but a reduced rate of water loss is the only possible mechanism to cope with drought stress in males. Finally, acclimated females showed a significant increase in drought resistance associated with higher trehalose content as well as dehydration tolerance, while males showed no acclimation response. Thus, sex-specific differences in desiccation resistance of D. kikkawai are associated with divergence in some water balance strategies, despite a lack of differences in the rate of water loss between the two sexes.

Rapid effects of humidity acclimation on stress resistance in Drosophila melanogaster.

We tested the hypothesis whether developmental acclimation at ecologically relevant humidity regimes (40% and 75% RH) affects desiccation resistance of pre-adults (3rd instar larvae) and adults of Drosophila melanogaster Meigen (Diptera: Drosophilidae). Additionally, we untangled whether drought (40% RH) acclimation affects cold-tolerance in the adults of D. melanogaster. We observed that low humidity (40% RH) acclimated individuals survived significantly longer (1.6-fold) under lethal levels of desiccation stress (0-5% RH) than their counter-replicates acclimated at 75% RH. In contrast to a faster duration of development of 1st and 2nd instar larvae, 3rd instar larvae showed a delayed development at 40% RH as compared to their counterparts grown at 75% RH. Rearing to low humidity conferred an increase in bulk water, hemolymph content and dehydration tolerance, consistent with increase in desiccation resistance for replicates grown at 40% as compared to their counterparts at 75% RH. Further, we found a trade-off between the levels of carbohydrates and body lipid reserves at 40% and 75% RH. Higher levels of carbohydrates sustained longer survival under desiccation stress for individuals developed at 40% RH than their congeners at 75% RH. However, the rate of carbohydrate utilization did not differ between the individuals reared at these contrasting humidity regimes. Interestingly, our results of accelerated failure time (AFT) models showed substantial decreased death rates at a series of low temperatures (0, -2, or -4°C) for replicates acclimated at 40% RH as compared to their counter-parts at 75% RH. Therefore, our findings indicate that development to low humidity conditions constrained on multiple physiological mechanisms of water-balance, and conferred cross-tolerance towards desiccation and cold stress in D. melanogaster. Finally, we suggest that the ability of generalist Drosophila species to tolerate fluctuations in humidity might aid in their existence and abundance under expected changes in moisture level in course of global climate change.

Invasion and adaptation of a warm-adapted species to montane localities: effect of acclimation potential.

Drosophila ananassae has successfully invaded the cold and dry montane localities of the Western Himalayas in recent years. The ability of this desiccation- and cold-sensitive tropical species to evolve in response to seasonal changes in montane localities is largely unknown. Here, we investigated how this sensitive species adapt to seasonally varying environmental conditions that are lethal to its survival. We observed change in the frequency of dark and light morphs of D. ananassae in five mid-altitude localities during the last decade (2000-2010). We document invasion of D. ananassae to montane localities and increase in frequency of the dark morph. The stress tolerance of morphs (dark and light) remained unaffected of developmental acclimation. However, adult acclimation has shown significant effects on tolerance to various environmental stresses in morphs and effect of this acclimation persist for long durations. Desiccation and cold stress tolerance was increased after adult acclimation for respective stress in the dark morph; while tolerance of the light morph was not affected. Further, heat tolerance of the light morph was increased after adult heat acclimation with no influence on heat tolerance of the dark morph. Our results suggest a possible role of adult acclimation in successful invasion and adaptation of D. ananassae to montane localities. Future experiments should be carried out to determine whether the survival in adverse conditions of low versus high temperature and humidity during seasonal changes is assisted by different acclimation abilities of the two morphs of D. ananassae.