Seema Ramniwas

Changes in body melanisation and desiccation resistance in highland vs. lowland populations of D. melanogaster.

Wild caught samples of Drosophila melanogaster from five highland localities showed parallel changes in melanisation and desiccation resistance in darker versus lighter phenotypes, i.e. darker flies (>45% melanisation) showed significantly higher desiccation resistance than lighter flies (<30% melanisation). In order to find an association between body melanisation and desiccation resistance, highland and lowland populations from tropical and subtropical regions (11.15-31.06 degrees N) of the Indian subcontinent were raised and investigated at 21 degrees C for four physiological traits, i.e. per cent body melanisation, desiccation resistance, rate of water loss and rate of water absorption. On the basis of mother-offspring regression, body melanisation and desiccation resistance showed higher heritability (0.58-0.68) and thus these traits are suitable for laboratory analyses. Significantly higher melanisation as well as desiccation resistance were observed in highland populations as compared with lowland populations. The rates of water loss as well as absorption were negatively correlated with body melanisation, i.e. darker flies from highlands showed a reduced rate of water loss as well as a lower rate of water absorption while the reverse trend was observed in lighter flies from lowlands. On the basis of multiple regressions, significant effects due to combined altitude and latitude were observed for all the four physiological traits. Local climatic conditions (i.e. annual average temperature and relative humidity) helped in explaining parallel changes in body melanisation and desiccation resistance in D. melanogaster.

Body melanization and its adaptive role in thermoregulation and tolerance against desiccating conditions in drosophilids

Melanism seems to have evolved independently through diverse mechanisms in various taxa and different ecological factors could be responsible for selective responses. Increased body melanization at higher altitudes as well as latitudes is generally considered to be adaptive for thermoregulation. Physiological traits such as body melanization and desiccation resistance have been investigated independently in diverse insect taxa at three levels: within populations, between populations and among species. A substantial number of Drosophila studies have reported clinal variations in both these traits along latitude. A possible link between these traits had remained unexplored in wild and laboratory populations of ectothermic insect taxa, including drosophilids, to date. Simultaneous analysis of these traits in assorted darker and lighter phenotypes in each population in the present study showed parallel changes for body melanization and desiccation resistance. The mechanistic basis of evolving desiccation resistance was explained on the basis of differential rates of water loss per hour in darker versus lighter phenotypes in six populations of Drosophila melanogaster from adjacent localities differing substantially in altitude all along the Indian subcontinent. Data on cuticular impermeability suggest a possible role of melanization in desiccation tolerance. However, substantial gaps remain in extending these results to other insect taxa and further exploring the physiological and molecular changes involved in melanization for conferring desiccation resistance.

Seasonal changes in humidity level in the tropics impact body color polymorphism and desiccation resistance in Drosophila jambulina - evidence for melanism-desiccation hypothesis.

Drosophila jambulina exhibits color dimorphism controlled by a single locus but its ecological significance is not clear. Dark and light morphs differ significantly in body melanisation, desiccation resistance, rate of water loss, mating activity and fecundity. Interestingly, this species lacks clinal variation for body size, desiccation resistance and life history traits. For body melanisation, lack of geographical variation as well as plastic effects is not in agreement with a thermal melanism hypothesis. However, based on field data, there are seasonal changes in phenotypic frequencies of dark and light body color morphs which correlate significantly with variation in humidity levels. Under short-term (8h) desiccation stress, we observed higher number of assortative matings, longer copulation period and increased fecundity for dark strains as compared with light strains. By contrast, both the morphs when exposed to high humid conditions exhibited higher assortative matings and fecundity for light strains as compared with dark strains. In tropical populations of D. jambulina, body color polymorphism seems to be maintained through humidity changes as opposed to thermal melanism. Thus, seasonal changes in the frequency of body color morphs in this tropical species supports melanism-desiccation hypothesis.

Impact of darker, intermediate and lighter phenotypes of body melanization on desiccation resistance in Drosophila melanogaster

Abstract A possible link between melanization and desiccation resistance can be inferred if within population differences in melanization find significant correlations with desiccation resistance and its mechanistic basis i.e. rate of water loss/hr. Accordingly, darker, intermediate and lighter phenotypes of body melanization were analyzed in wild and laboratory reared Drosophila melanogaster L. (Diptera: Clyclorrapha) populations from highland and lowland sites located in close proximity at five different latitudinal locations (11.15 °N to 31.06°N) within the Indian subcontinent. In large population samples, occurrence of significant within population variability made it possible to assort non-overlapping phenotypes of body coloration (i.e. lighter (< 25%), intermediate (30 to 40%) and darker (> 45%)) for all the populations which were further investigated for desiccation resistance and rate of water loss/hr. Significantly, higher desiccation resistance but much reduced rate of water loss/hr were observed in darker and intermediate phenotypes in all the populations. By contrast, lighter phenotypes exhibited lower desiccation tolerance but higher rate of water loss/hr. A regression analysis between traits provided similar slope values for wild and laboratory populations. For all three physiological traits, predicted trait values from multiple regression analysis as a simultaneous function of annual average temperature and relative humidity, matched the observed values. We infer that parallel changes in melanization and desiccation resistance may result from decreasing annual average temperature and relative humidity along increasing latitude as well as altitude on the Indian subcontinent.

Variations in body melanisation, ovariole number and fecundity in highland and lowland populations of Drosophila melanogaster from the Indian subcontinent

We investigated geographical variations in three fitness‐related traits (body melanisation, ovariole number and fecundity) in laboratory‐reared offspring of 10 populations of Drosophila melanogaster. The populations were collected from adjacent lowland and highland localities (∼80–100 km apart) in the tropical as well as subtropical regions (11.15–31.06 °N) covering a linear distance about 3 000 kilometers from south to north on the Indian subcontinent. Persistence of within‐as well as between‐population differences at 21 °C suggest that observed variations in fitness‐related traits have a genetic basis. Populations from higher altitudes showed consistently higher trait values (1.4‐fold increase) as compared with their corresponding lowland populations. By contrast, latitudinal variations were about two‐fold higher across the entire continent. Along latitude as well as altitude, population means showed higher correlation values (r > 0.98) between all the three fitness traits. However, on the basis of within‐population analysis (assorted darker and lighter flies), changes in body melanisation were significantly correlated with fecundity but not with ovariole number. Thus, analysis of within‐population trait variability should be preferred as compared with data on population means for adaptive significance of fitness‐related traits. In the present study, the role of climatic selection is evident from regression analysis with changes in annual average temperature of the sites of origin of populations along latitude as well as altitude.

Direct and correlated responses to laboratory selection for body melanisation in Drosophila melanogaster: support for the melanisation–desiccation resistance hypothesis

SUMMARY For Drosophila melanogaster, cuticular melanisation is a quantitative trait, varying from no melanin to completely dark. Variation in melanisation has been linked with stress resistance, especially desiccation, in D. melanogaster and other species. As melanism has a genetic component, we selected melanic and non-melanic phenotypes of D. melanogaster in order to confirm the association of desiccation resistance and rate of water loss with cuticular melanisation previously reported for this species. A bidirectional selection experiment for dark (D1–D4) and light (L1–L4) body colour in D. melanogaster was conducted for 60 generations. We observed a 1.6-fold increase in abdominal melanisation in selected dark strains and a 14-fold decrease in selected light strains compared with control populations. Desiccation resistance increased significantly in the dark-selected morphs as compared with controls. The observed increase in desiccation resistance appeared as a consequence of a decrease in cuticular permeability. Our results show that traits related to water balance were significantly correlated with abdominal melanisation and were simultaneously selected bidirectionally along with melanisation.

Fitness consequences of body melanization in Drosophila immigrans from montane habitats.

We investigated eight populations of Drosophila immigrans from low to high montane localities (600–2202 m) for altitudinal variations in abdominal melanization and fitness‐related traits (desiccation resistance, copulation duration, and fecundity). On the basis of common garden experiments, persistence between‐population differences at 21°C suggests that observed variations in fitness‐related traits have a genetic basis. Parent–offspring regression analyses showed higher heritability (h2= 0.77) for melanization patterns on all the abdominal tergites. All the traits showed significantly higher repeatability across generations. Under colder and drier environments in dispersed montane localities, abdominal melanization and desiccation resistance significantly increased (1.5–1.9 fold) along altitude. Thus, there are correlated effects of abdominal melanization on desiccation resistance. Genetic correlations, based on family means, were significantly high between abdominal melanization and other fitness traits. Furthermore, darker flies along increasing altitude resulted in a 35–40% increase in copulation duration as well as fecundity. There are significantly positive correlations of abdominal melanization with copulation duration as well as fecundity on the basis of within‐ as well as between‐population variations. Such observations are in agreement with the thermal budget hypothesis. Present data suggest that changes in body melanization impact fitness‐related traits in montane populations of Drosophila immigrans.

Divergence of desiccation-related traits in two Drosophila species of the takahashii subgroup from the western Himalayas

SUMMARY Drosophila nepalensis is more abundant under colder and drier montane habitats in the western Himalayas compared with Drosophila takahashii, but the mechanistic basis of such a climatic adaptation is largely unknown. We tested the hypothesis that divergence in the physiological basis of desiccation-related traits is consistent with species-specific adaptations to climatic conditions. Drosophila nepalensis showed approximately twofold higher desiccation resistance, hemolymph content as well as carbohydrate content than D. takahashii despite a modest difference in rate of water loss (0.3% h–1). Water loss before succumbing to death (dehydration tolerance) was much higher in D. nepalensis (82.32%) than in D. takahashii (∼50%). A greater loss of hemolymph water under desiccation stress until death is associated with higher desiccation resistance in D. nepalensis. In both species, carbohydrates were utilized under desiccation stress, but a higher level of stored carbohydrates was evident in D. nepalensis. Further, we found increased desiccation resistance in D. nepalensis through acclimation whereas D. takahashii lacked such a response. Thus, species-specific divergence in water-balance-related traits in these species is consistent with their adaptations to wet and dry habitats.

Climate change, boundary increase and elongation of a pre‐existing cline: A case study in Drosophila ananassae

During the past two to three decades, Drosophila ananassae, a warm adapted tropical species, has invaded low to mid altitude localities in the western Himalayas. Due to its cold sensitivity, this species had never been recorded from higher latitudes as well as altitudes in India to the 1960s. A latitudinal cline in this desiccation‐sensitive species corresponds with southern humid tropical localities rather than northern drier subtropical localities. An extension of its cline into lowland to midland montane localities has resulted due to global climatic change as well as local thermal effects through anthropogenic impact. However, D. ananassae populations at species borders are characterized by lower genetic variability for body melanization as well as for desiccation resistance. There is a lack of thermal plastic effects for body melanization, and the observed extended cline might represent evolutionary (genetic) response due to selection pressure imposed by drier habitats. A comparison of fecundity, hatchability and viability at three growth temperatures (17, 20 and 25°C) showed significant reduction in trait values at 17°C in D. ananassae. Thus, its recent range expansion into northern montane localities might involve genetic effects on stress‐related traits and plastic effects on life history traits. We suggest that D. ananassae could serve as an indicator species for analyzing range expansion under changing climatic conditions.

Divergent strategy for adaptation to drought stress in two sibling species of montium species subgroup: Drosophila kikkawai and Drosophila leontia

Drosophila leontia (warm adapted) has been considered as a sister species of Drosophila kikkawai (sub-cosmopolitan) with a very similar morphology. We found divergent strategies for coping with desiccation stress in these two species of montium subgroup. Interestingly, in contrast to clinal variation for body melanization in D. kikkawai, cuticular lipid mass showed a positive cline in D. leontia across a latitudinal transect. On the basis of isofemale line analysis, within population trait variability in cuticular lipid mass per fly is positively correlated with desiccation resistance and negatively correlated with cuticular water loss in D. leontia. A comparative analysis of water budget of these two species showed that higher abdominal melanization, reduced rate of water loss and greater dehydration tolerance confer higher desiccation resistance in D. kikkawai while the reduced rate of water loss is the only possible mechanism to enhance desiccation tolerance in D. leontia. The use of organic solvents has supported water proofing role of cuticular lipids in D. leontia but not in D. kikkawai. Thus, we may suggest that body melanization and cuticular lipids may represent alternative strategies for coping with dehydration stress in melanic versus non-melanic drosophilids. In both these species, carbohydrates were utilized under desiccation stress but a higher level of stored carbohydrates was evident in D. kikkawai. Further, we found increase desiccation resistance in D. kikkawai through acclimation while D. leontia lacks such a response. Thus, species specific divergence in water balance related traits in these species are consistent with their adaptations to wet and dry habitats.