Heat-induced hormesis in longevity is linked to heat-stress sensitivity across laboratory populations from diverse altitude of origin in Drosophila buzzatii.

Heat-induced hormesis in longevity is the increase in life span resulting from the previous exposure to a mild heat stress early in life. Here we examined heat-induced hormesis of Drosophila buzzatii in five mass-mating populations, which were derived from five wild populations along an elevation gradient from 202 to 1855 m above sea level in North-Western Argentina. Five day old flies were exposed to 37.5 °C for 90 min to induce hormesis and its possible variation across altitudinal populations. This heat treatment strongly extended longevity in lowland-derived flies from the most heat-resistant population only. Both heat-induced effects on longevity and heat-knockdown time (heat-stress sensitivity) were negatively correlated to altitude of population of origin. Hormesis was positively correlated to heat-knockdown time across populations. These results indicate that variation in heat-induced hormesis can not be considered as independent of heat-stress sensitivity (or heat-knockdown time) in populations of insects.

Mating success at elevated temperature is associated to thermal adaptation in a set of recombinant inbred lines of Drosophila melanogaster.

In insects, mating ability at elevated temperature can be relevant for adaptation to heat-stressed environments and global warming. Here, we examined copulation latency (T1), copulation duration (T2), and mating frequency (T3, an index of mating success) in two related sets of recombinant inbred lines (RIL) in Drosophila melanogaster at both elevated (33 °C) and benign (25 °C) temperatures. One of these RIL sets (RIL-SH2) was shown to be consistently more resistant in both heat knockdown and heat-shock survival assays than its related set (RIL-D48) in previous studies. Negative correlations across RILs were found between T1 and T3 in this study. Flies from the heat-resistant set of RIL (RIL-SH2) were better able to mate at elevated temperature than flies from the heat-susceptible set (RIL-D48). Quantitative trait locus (QTL) mapping identified temperature-dependent QTLs for all traits (T1, T2 and T3) on all the three major chromosomes. Mating success at elevated temperature was found to be influenced by multiple QTLs. At elevated temperature, several QTLs for mating traits co-localized with QTLs that were previously associated with thermotolerance. The genetic basis for T1, T2 and T3 at the elevated temperature was found to be largely different from the genetic basis controlling the variation for mating success at benign temperature, as there was only a very low (or even null) number of QTLs overlapping across temperatures.

Genetic variation in the heat-stress survival of embryos is largely decoupled from adult thermotolerance in an intercontinental set of recombinant lines of Drosophila melanogaster.

In insects, thermal adaptation works on the genetic variation for thermotolerance of not only larvae and adults but also of the immobile stages of the life cycle including eggs. In contrast to adults and larvae, the genetic basis for thermal adaptation in embryos (eggs) remains to be tested in the model insect Drosophila melanogaster. Quantitative-trait loci (QTL) for heat-stress resistance in embryos could largely differ from previously identified QTL for larvae and adults. Here we used an intercontinental set of recombinant inbred lines (RIL), which were previously used to identify thermotolerance-QTLs in adults and larvae because of their high variation segregating for adult thermotolerance. Eggs appeared to be more heat resistant than larvae and adults from previous studies on these RIL, though different heat-shock assays were used in previous studies. We found that variation in thermotolerance in embryos can be, at least partially, genetically decoupled from thermotolerance in the adult insect. Some RIL that are heat resistant in the adult and larvae can be heat susceptible in embryos. Only one small-effect QTL out of five autosomal QTL co-localized between embryo and other ontogenetic stages. These results suggest that selection for thermal adaptation in adult flies and larvae is predicted to have only a small impact on embryo thermotolerance. In addition, heat-stress tolerance of insects can be measured across ontogenetic stages including embryos in order to better predict thermal adaptive limits of populations and species.

Heat knockdown resistance and chill-coma recovery as correlated responses to selection on mating success at high temperature in Drosophila buzzatii.

Reproduction and related traits such as mating success are strongly affected by thermal stress. We tested direct and correlated responses to artificial selection in replicated lines of Drosophila buzzatii that were selected for mating success at high temperature. Knockdown resistance at high temperature (KRHT) and chill-coma recovery (CCR) were tested as correlated selection responses. Virgin flies were allowed to mate for four hours at 33°C in three replicated lines (S lines) to obtain the selected flies and then returned at 25°C to lay eggs. Other three replicated lines were maintained at 25°C without any selection as control (C lines). After 15 selection generations, KRHT and CCR were measured. Both traits were assessed in flies that did not receive any hardening pretreatments as well as in flies that were either heat or cold hardened. Thermotolerance traits showed significant correlated responses with higher KRHT in S than in C lines, both with a heat-hardening pretreatment and without a heat-hardening pretreatment. CCR time was longer in S than in C lines both with a cold-hardening pretreatment and without a cold-hardening pretreatment. Hardening treatments improved both KRHT and CCR in all cases excepting KRHT in C lines. Overall, KRHT and CCR showed an antagonistic pattern of correlated responses to our selection regime, suggesting either pleiotropy or tightly linked trait-specific genes partially affecting KRHT and CCR.

Negative genetic correlation between longevity and its hormetic extension by dietary restriction in Drosophila melanogaster.

Longevity is a highly malleable trait which is influenced by many genetic and environmental factors including nutrition. Mild stress of dietary restriction (DR) is often beneficial by extending longevity in many organisms. Here, DR-induced effects on longevity were tested for genetic variation in a set of recombinant inbred lines (RIL) in D. melanogaster. Genetic variability was significant in the longevity response following a DR-treatment across RIL, with detrimental effects in several RIL but beneficial effects in other RIL. One quantitative trait locus (QTL) was consistently significant in the middle of chromosome 2 for DR-induced changes in longevity, including hormesis (an increase in longevity by DR). Another QTL co-localized with a previously found QTL for starvation resistance in females. Several other QTL were also significant on most chromosomal arms. Longevity in controls was negatively correlated to DR effects across RIL for longevity in females, the sex showing higher DR-induced hormesis. This negative genetic correlation highlights the importance to further investigate the effects of genetic variation in the strength of DR-induced hormesis in longevity and its sex-specificity.

Elevated extension of longevity by cyclically heat stressing a set of recombinant inbred lines of Drosophila melanogaster throughout their adult life.

An extremely high (about 100 %) increase in longevity is reported for a subset of recombinant inbred lines (RILs) of Drosophila melanogaster subjected to a cyclic heat stress throughout the adult life. Previous work showed that both longevity and heat sensitivity highly differed among RILs. The novel heat stress treatment used in this study consisted of 5 min at 38 °C applicated approximately every 125 min throughout the adult life starting at the age of 2 days. In spite of the exceptionally high increase in longevity in a set of RILs, the same heat stress treatment reduced rather than increased longevity in other RILs, suggesting that heat-induced hormesis is dependent on the genotype and/or the genetic background. Further, one quantitative trait locus (QTL) was identified for heat-induced hormesis on chromosome 2 (bands 28A1-34D2) in one RIL panel (RIL-D48) but it was not significant in its reciprocal panel (RIL-SH2). The level of heat-induced hormesis showed a sexual dimorphism, with a higher number of lines exhibiting higher hormesis effects in males than in females. The new heat stress treatment in this study suggests that longevity can be further extended than previously suggested by applying a cyclic and mild stress throughout the life, depending on the genotype.

Effects of dietary composition on life span of Drosophila buzzatii and its short-lived sibling species D. koepferae.

Two sibling Drosophila species dramatically divergent in longevity, Drosophila buzzatii and D. koepferae, were examined for possible effects of both developmental culture medium and dietary composition (DC) on longevity. Longevity was greatly increased in the longer lived D. buzzatii when flies were reared and fed on a rich-in-nutrient and cactus-based culture (R-CBC) as compared to longevity in a poor nutrient culture (PNC). In D. buzzatii, life span was further increased by exposing flies to short periods of a poor-in-nutrient and cactus-based culture (P-CBC). In contrast, variation in the here used nutrient composition did not change life span in the shorter lived D. koepferae, as longevity in this species did not differ among R-CBC, P-CBC and PNC cultures. Hormesis is a plausible explanation for the beneficial biological effects against aging arising from brief exposure to a lowed calorie food source in D. buzzatii. This study shows that genetic variation between closely related species is substantial for dietary effects on longevity.

Survival of heat stress with and without heat hardening in Drosophila melanogaster: interactions with larval density.

Survival of a potentially lethal high temperature stress is a genetically variable thermal adaptation trait in many organisms. Organisms cope with heat stress by basal or induced thermoresistance. Here, we tested quantitative trait loci (QTL) for heat stress survival (HSS) in Drosophila melanogaster, with and without a cyclic heat-hardening pre-treatment, for flies that were reared at low (LD) or high (HD) density. Mapping populations were two panels of recombinant inbred lines (RIL), which were previously constructed from heat stress-selected stocks: RIL-D48 and RIL-SH2, derived from backcrosses to stocks of low and high heat resistance, respectively. HSS increased with heat hardening in both LD and HD flies. In addition, HSS increased consistently with density in non-hardened flies. There was a significant interaction between heat hardening and density effects in RIL-D48. Several QTL were significant for both density and hardening treatments. Many QTL overlapped with thermotolerance QTL identified for other traits in previous studies based on LD cultures only. However, three new QTL were found in HD only (cytological ranges: 12E-16F6; 30A3-34C2; 49C-50C). Previously found thermotolerance QTL were also significant for flies from HD cultures.

Quantitative trait loci for longevity in heat-stressed Drosophila melanogaster.

Longevity is a typical quantitative trait which is influenced by multiple genes. Here we explore the genetic variation in longevity of Drosophila melanogaster in both mildly heat-stressed and control flies. Quantitative trait loci (QTL) analysis for longevity was performed in a single-sex environment at 25°C with and without a mild heat-stress pre-treatment, using a previously reported set of recombinant inbred lines (RIL). QTL regions for longevity in heat-stressed flies overlapped with QTL for longevity in control flies. All longevity QTL co-localized with QTL for longevity identified in previous studies using very different sets of RIL in mixed sex environments, though the genome is nearly saturated with QTL for longevity when considering all previous studies. Heat stress decreased the number of significant QTL for longevity if compared to the control environment. Our mild heat-stress pre-treatment had a beneficial effect (hormesis) more often in shorter-lived than in longer-lived RIL.

Quantitative trait locus for starvation resistance in an intercontinental set of mapping populations of Drosophila melanogaster.

Starvation resistance (SR) is an important trait for survival of insects in the wild. We used recombinant inbred lines (RIL) to search for quantitative trait loci (QTL) in crosses between intercontinental inbred lines that were originally selected for heat-knockdown resistance. SR was measured as the time of survival under repeated events of starvation. SR was consistently higher in females than in males. Composite interval mapping identified one QTL region (cytological range 64D-66E2) on the left arm of chromosome 3 in males, and no QTL was found in females. Many candidate genes that were identified in previous studies of QTL for stress resistance are included within this QTL region. The QTL-allele that decreased SR was found in the line originating from the colder population (Denmark). We discuss our results with regard to multiple candidate genes, noncolocalization with thermotolerance QTL, and possible geographical variation.

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.

Developmental time and size-related traits in Drosophila buzzatii along an altitudinal gradient from Argentina.

Clinal analysis for fitness-related traits provides a well-known approach to investigate adaptive evolution. Several fitness-related traits (developmental time, thorax length, wing length and wing loading) were measured at two laboratory generations (G7 and G33) of D. buzzatii from an altitudinal gradient from northwestern Argentina, where significant thermal differences persist. Developmental time (DT) was positively correlated with altitude of origin of population. Further, DT was negatively correlated with maximal mean temperature at the site of origin of population, and this thermal variable decreases with altitude. Wing loading tended to be larger in highland than in lowland populations, suggesting that flight performance is subject to stronger selection pressure in highland populations. Developmental time showed a significant increase with laboratory generation number. There was no significant correlation between developmental time and body size across populations along the altitudinal cline of DT. This result illustrates that developmental time and body size do not always evolve in the same direction, even though both traits are often positively and genetically correlated in a well-known tradeoff in Drosophila.