Different laboratory strains of Drosophila melanogaster are reported to differ considerably in their physiology, behavior, and life-histories, due to their adaptations to different laboratory conditions. Recent advances in insect aging research have highlighted the importance of protein:carbohydrate (P:C) balance as a key dietary determinant of lifespan and other components of fitness, but it remains unexplored whether P:C balance affects the fitness-related traits of D. melanogaster in a strain-specific manner. The purpose of this study was to compare the life-history consequences of six different laboratory strains of D. melanogaster (three Canton-S substrains, w1118, yw, and Oregon-R) allocated to four synthetic diets differing in P:C ratio (1:16, 1:4, 1:1, or 4:1). Five components of fitness (lifespan, fecundity, larval viability, development time, and body mass) were recorded from flies maintained at 25oC under L:D 12:12 photoperiod. All strains exhibited qualitatively similar responses to dietary P:C balance, with the increase in P:C ratio being associated with shortened lifespan and improved egg production. In all strains, fly larvae confined to P:C 1:16 suffered high mortality, retarded growth, and reduced body size. As indicated by significant diet×strain interactions for all measured fitness components, the magnitude of such diet effect varied among different laboratory strains in D. melanogaster. Possible explanations for such strain differences are discussed.

Diet and temperature are the two most critical environmental factors affecting life-history traits in insects, but the combined effects of these factors have been rarely investigated. In this study, various life-history traits were recorded from adult and larval Drosophila melanogaster fed on one of eight synthetic diets differing in protein:carbohydrate ratio (P:C=1:16, 1:8, 1:4, 1:2, 1:1, 2:1, 4:1, or 8:1) under one of six ambient temperatures (13, 18, 23, 28, 31, or 33oC). The patterns of adult and larval life-history traits expressed across 48 diet-by-temperature combinations were visualized using thin-plate spline technique and the presence of any significant linear, quadratic, and correlational effects of diet and temperature on trait expressions was analyzed using a second-order polynomial multiple regression. Life-history traits exhibited qualitatively different responses to variations in both diet and temperature, with the maximal expression of each trait being achieved at a completely divergent region of the diet-temperature fitness landscape. In adult females, for example, lifespan was maximized at P:C 1:16 under 13oC, but fecundity was maximized at P:C 4:1 under 28oC. These results provide empirical support for the emerging notion that environmental factors, such as diet and temperature, can mediate life-history trade-offs in insects.

과일이나 농작물의 부패 및 발효 환경에서는 Methanol, Ethanol, Acetic acid을 비롯한 다양한 화학물질들이 생산된다. Drosophila melanogaster는 이러한 발효·부패 환경에 서식하면서 일정 농도 이상의 다양한 화학물질에 지속적으로 노출되어 생존하도록 적응되어온 것으로 생각된다. 다양한 화학물질이 포함한 환경에 안정적으로 서식하기 위해서는 D. melanogaster는 화학물질에 능동적으로 반응하여 해독 유전자나 대사 관련 유전자의 발현량을 변화 시킴으로써 발효·부패 환경에서 생성되는 화학물질에 대한 높은 내성을 가지고 있을 것으로 판단된다. 현재까지 유전자의 발현량 측정을 위해 real-time PCR를 이용하여 reference gene의 발현량을 기준으로 정량화하는 방법이 가장 널리 사용되고 있다. 그러나 조직별, 환경별, 발달단계를 비롯한 다양한 조건에서 안정적으로 발현되는 reference 유전자 선정이 필수적으로 선행되어야 하므로 본 연구에서는 발효·부패 환경에서 생산되는 두 화학물질인 Methanol과 Ethyl Acetate에 노출된 D. melanogaster에서 안정적으로 발현되는 reference gene을 찾는 연구를 실시하였다. 본 연구에서는 다양한 농도의 Methanol과 Ethyl Acetate을 D. melanogaster에 노출시킨 후 RNA 추출과 cDNA 합성을 실시였고, 5가지 후보 reference gene (hsp22, nd, rpL18, tbp and ef-1b)의 안정적 발현 여부를 qRT-PCR을 통해 조사하였으며, 유전자 발현의 안정성을 측정하는 3가지 프로그램(geNorm, NormFinder, BestKeeper)을 이용해 비교·분석하였다. 본 학회에서는 연구의 과정과 그 결과를 발표하고자 한다.

4,4’-dichlorodiphenyltrichloroethane (DDT) has been re-recommended by the World Health Organization for malaria mosquito control in Africa. Previous DDT use has resulted in predisposition of resistance, and with continued use resistance will increase further in terms of level and extent. Drosophila melanogaster is a model dipteran that has many available genetic tools, has been widely used for elucidating insecticide resistance mechanisms, and is related to malaria mosquitoes allowing for extrapolation. The 91-R strain of D. melanogaster is highly resistant to DDT (>1500-fold); however, there is no mechanistic scheme that accounts for this level of resistance. Recently, reduced penetration, increased detoxification, and direct excretion have been identified as resistance mechanisms in the 91-R strain. Their interactions, however, remain unclear. Use of Gal4/UAS-RNAi transgenic lines of D. melanogaster allowed for the targeted knockdown of genes putatively involved in DDT resistance and has identified the role of several cuticular proteins (Cyp4g1 and Lcp1), cytochrome P450 monooxygenases (Cyp6g1 and Cyp12d1), and ATP binding cassette transporters (mdr50, mdr65, and mrp1) in increased sensitivity to DDT. These findings have been further validated in 91-R flies using a nanoparticle-enhanced RNAi strategy, directly implication these genes in DDT resistance in 91-R flies.

Diets influence lifespan and reproduction in insects, but little is known how temperature modulates the impacts of diet on these two key fitness components. Here we examined the interactive effects of temperature and nutrient balance on lifespan and egg production rate in Drosophila melanogaster. Newly emerged adult D. melanogaster were allowed to feed ad libitum on one of eight chemically defined diets differing in P:C ratio (1:16, 1:8, 1:4, 1:2, 1:1, 2:1, 4:1, or 8:1) under one of six ambient temperatures (13, 18, 23, 28, 31, or 33℃). For both males and females, lifespan was longest for D. melanogaster fed on P:C 1:16 diet at 13℃ and shortened as both temperature and P:C ratio increased. As indicated by a significant temperature-by-diet interaction for lifespan, the diet effects on lifespan were more pronounced at lower temperatures than at higher temperatures. Egg production rate was maximized on P:C 4:1 diet at 28℃.

Protein and carbohydrate are two major macronutrients that exert profound influences over fitness in many insects, including Drosophila melanogaster. Until recently, most studies examining the impacts of these macronutrients on various life-history traits in this species have used semi-synthetic diets that are not nutritionally well-defined. Here we used chemically defined diets to examine the patterns of larval and adult traits expressed across 34 diets systematically varying in the ratio and concentration of protein and carbohydrate. The shapes of the nutritional landscapes plotted for all larval and adult traits differed significantly from one another. Diverging nutritional optima identified for these landscapes suggest that D. melanogaster cannot maximize the expression of all life-history traits simultaneously, thus leading them to face a nutrient-dependent life-history trade-off.

Temperature can modulate how insects respond to environmental stressors, such as starvation. In this study, we examine whether and how the effects of temperature on starvation resistance depend on nutritional condition and developmental stages in Drosophila melanogaster. Starvation resistance decreased as the temperature exposed during starvation rose from 18 to 28 ̊C, which was mainly caused by warming-induced increase in energy expenditure. When exposed to warm temperatures during feeding, D. melanogaster accumulated more energy reserves and thus become more starvation resistant. The temperature experienced during the larval stage also had a significant effect on starvation resistance at adult stages, with those larvae raised at cold temperatures developing into adult phenotypes with reduced resistance to starvation. This study suggests that the effects of temperature on starvation resistance are highly complex and context dependent in D. melanogaster.

Drosophila melanogaster and Drosophila suzukii are the species of the family Drosophilidae. Although these two fruit flies are taxonomically close species, D. suzukii is thought to be evolutionally adapted to the flesh or maturing fruits, whereas D. melanogaster is adapted to more fermented environments. According to the previous studies, several environmental toxins, such as acetic acid, ethanol, methanol and phenylacetate, ect., have been identified from rotten fruit and fermentation procedures. Considering the differences of distinct habitat between two flies, D. melanogaster is hypothesized to exhibit higher tolerance to the chemical toxins than D. suzukii. Therefore, in this study, we compared the tolerance and susceptibility of two fruit flies to three chemicals (acetic acid, ethanol, 2-phenlyethano).

Temperature can affect the ability of insects to tolerate prolonged period of food deprivation through altering the amountof energy storage, the speed of energy expenditure, or the threshold energy storage for survival. In this study, we examinedthe mechanistic basis of the temperature-dependence of starvation resistance in Drosophila melanogaster. Starvation resistancedecreased as the temperature experienced during starvation rose from 18 to 28 ̊C. This warming-mediated decrease instarvation resistance was due to accelerated energy expenditure. However, the threshold energy storage for survival wasnot affected by starvation temperature. Exposure to warm temperatures during feeding led D. melanogaster to accumulatemore energy reserves and thus to become more starvation resistant. This study highlights the important role played bytemperature in shaping the phenotypic responses of insects to starvation.

In many insects, female receives a large amount of sperm and ejaculates upon copulation, and keeps them in her uterus for some time, during which she stores sperm. After varying delays, the female removes unused sperm and ejaculates through sperm ejection behavior, which is known as a major mechanism for the post-copulatory sexual selection. In the fruitfly Drosophila melanogaster, timing of sperm ejection behavior is controlled by the diuretic hormone 44 (Dh44) pathway. To identify pathways that regulate Dh44 activities and sperm ejection, here we examine effects of olfactory cues. We will present our recent results suggesting that male odors can be one of upstream signaling pathway that modulates a brain neuropeptide pathway.

Acetylcholinesterase (AChE) is an enzyme for hydrolyzing neurotransmitter acetylcholine. Soluble form of AChE is generated via alternative splicing and functions as a bioscavenger in Dropsophila melanogaster. In this study, effects of ethanol and acetic acid on the soluble AChE expression were investigated. Treatment of ethanol and acetic acid results in over-expression of soluble AChE in the abdomen in a dose-dependent manner. However, no apparent change in AChE expression was observed in the head. Our finding suggests that the soluble AChE is involved in chemical defense against high concentration of ethanol, which is a common by-product from fermented food，and acetic acid, the main metabolite of ethanol. Thus, high level of ethanol and acetic acid resistance in D. melanogaster appears to be evolved via the induction mechanism of soluble AChE expression.

노랑초파리(Drosophila melanogaster)는 가장 연구가 많이 되어져 있는 모델동물로서 다양한 연구에 활용이 되어져 왔다. 하지만, 작물보호제에 대한 약제 저항성 모델로는 활용이 되어지지 않고 있다. 우리는 노랑초파리의 수컷에 Ethylene methane sulfonate를 처리하여 전체염색체에 돌연변이를 유도 한 후에 Chlorantraniliprole을 처리하여 약제에 대하여 저항성을 가지는 초파리 계통을 제작, 확립하였다. 저항성 계통에 대한 전체발현 유전체 분석과 micro-RNA분석을 수행하여 발현이 변화된 유전자를 밝혀내고 돌연변이 유전자를 발굴하는 연구를 수행하고 있다. 이러한 연구는 저항성 관련 기전을 밝혀내는 것과 동시에 내성에 관련된 기전을 밝혀내는데도 커다란 도움을 줄 수 있을 것이다. (본 연구는 농촌진흥청 협동연구사업 “초파리를 이용한 교차저항성 발달 모델 및 기작 구명(PJ01082103)”으로 수행되었음).

Acetylcholinesterase (AChE) is a hydrolase that hydrolyzes the neurotransmitter acetylcholine. Soluble form of AChE is generated via alternative splicing and functions as a bioscavenger in Dropsophila melanogaster. In this study, effects of acetic acid on the soluble AChE expression were investigated. Treatment of acetic acid resulted in over-expression of soluble AChE in the abdomen in a dose-dependent manner. The soluble AChE was determined to be expressed in the fat body. However, no apparent change in AChE expression was observed in the head. Our finding suggests that the soluble AChE is involved in chemical defense against high concentration of acetic acid, which is a common by-product in fermenting foods. The high level of acetic acid resistance in D. melanogaster, thus, appears to have been evolved via the induction mechanism of soluble AChE expression.

Recent studies have shown that mating can alter starvation resistance in female D. melanogaster, but little is known about the behavioral and physiological mechanisms underlying such mating-mediated changes in starvation resistance. In the present study, we first investigated whether the effect of mating on starvation resistance is sex-specific in D. melanogaster. As indicated by a significant sex × mating status interaction, mating increased starvation resistance in females but not in males. In female D. melanogaster, post-mating increase in starvation resistance was mainly attributed to increases in food intake and in the level of lipid storage relative to lean body weight. We then performed quantitative genetic analysis to estimate the proportion of the total phenotypic variance attributable to genetic differences (i.e., heritability) for starvation resistance in mated male and female D. melanogaster. The narrow-sense heritability (h2) of starvation resistance was 0.235 and 0.155 for males and females, respectively. Mated females were generally more resistant to starvation than males, but the degree of such sexual dimorphism varied substantially among genotypes, as indicated by a significant sex × genotype interaction for starvation resistance. Cross-sex genetic correlation was greater than 0 but less than l for starvation resistance, implying that the genetic architecture of this trait was partially shared between the two sexes. For both sexes, starvation resistance was positively correlated with longevity and lipid storage at genetic level. The present study suggests that sex differences in starvation resistance depend on mating status and have a genetic basis in D. melanogaster.

Macronutrient balance has a strong influence on fitness in insects. Previous studies have revealed that altering the concentrations of yeast and sugar in the semi-synthetic diet has a profound impact on lifespan and fecundity in Drosophila melanogaster, indicating the role of dietary protein:carbohydrate (P:C) balance in determining these two key components of fitness. However, since yeast contains not only proteins but also other macro- and micronutrients, this lifespan-determining role of dietary P:C balance needs to be corroborated using a chemically defined diet. In this study, the effects of dietary P:C balance on lifespan and fecundity were investigated in female D. melanogaster flies on one of eight isocaloric synthetic diets differing in P:C ratio (0:1, 1:16, 1:8, 1:4, 1:2, 1:1, 2:1 or 4:1). Lifespan and dietary P:C ratio were related in a convex manner, with lifespan increasing to a peak at the two intermediate P:C ratios (1:2 and 1:4) and falling at the imbalanced ratios (0:1 and 4:1). Ingesting nutritionally imbalanced diets caused flies to start ageing earlier and senesce faster. Egg production increased progressively as the dietary P:C ratio rose from 0:1 to 4:1. Long-lived flies at the intermediate P:C ratios(1:2 and 1:4) stored a greater amount of lipids than those short-lived ones at the two imbalanced ratios (0:1 and 4:1). These findings provide a strong support to the notion that dietary P:C balance is a critical determinant of lifespan and fecundity in D. melanogaster.

Internal sperm storage after mating is important for insect reproduction, because it permits delayed fertilisation, and post-copulatory mate choice in polyandrous females. The polyandry is common in many animal taxa including insects, because it increases female fitness by reducing the risk of infertility and providing opportunities for sperm competition and choice. The reproductive success of males, on the other hand, often depends upon avoidance of sperm competition by preventing mated females from copulating and receiving sperm from other male suitors. A widespread strategy used by males is the use of the male seminal fluid proteins (SFPs) that form the mating plug and alter female behaviors, for example by suppressing mating receptivity and elevating egg-laying. Under these circumstances, females are expected to evolve mechanism(s) to control exposure to the male SFPs in order to maximize fitness by balancing the positive and negative impacts of polyandry. Here, we discover that Drosophila melanogaster females eject male ejaculates 1-6 h after mating with a stereotypic behaviour, and that this is regulated by a brain neuropeptide pathway composed of diuretic hormone 44 (Dh44), and its receptor Dh44R1. We showed that suppressing Dh44 or Dh44R1 signals in the brain expedites sperm ejection, whereas enhancing Dh44 or Dh44R1 signals delays sperm ejection. This study uncovers a molecular mechanism by which females can influence sperm competition and selection, and counter actively the negative impact of polyandry.

Mating elicits a dramatic changes in physiology, behavior, and life-history traits in insects, but little is known about the relationship between mating and the capacity of insects to resist environmental stressors. Starvation is one of the most ubiquitous forms of environmental stress faced by all insects under natural conditions. Previous studies using Drosophila melanogaster flies has shown that mated females lived longer under starvation than did virgin females, but the mechanistic basis for such post-mating increase in starvation resistance remains largely unexplored. The objective of this study was to investigate the behavioral and physiological mechanisms of mating-induced alteration in starvation resistance and its heritable genetic variations in D. melanogaster. In the first experiment (Experiment 1), we compared starvation resistance (measured as starving time before death), body compositions, and food intake between mated and unmated flies of both sexes using a large outbred population. In the second experiment (Experiment 2), starvation resistance and body composition were quantified for mated male and female flies derived from each of 19 highly inbred genetic lines. Results from Experiment 1 showed that mated females were better able to resist starvation than virgin females and males because they ate more and thus laid down more fats in their body. Results from Experiment 2 revealed a significant heritable genetic variation in starvation resistance and its correlated body composition parameters for both sexes. Overall, females had a higher starvation resistance than males, but the magnitude of such intersexual difference varied among genetic lines, as suggested by a significant sex-by-line interaction. Cross-sex genetic correlations were highly significant and positive for starvation resistance, indicating that the genetic factors controlling the starvation resistance in D. melanogaster are shared between the two sexes.

Starvation resistance is an important fitness trait that is controlled by both environmental and heritable factors. The main objective of this study is to explore the genotype-by-nutrient interactions for starvation resistance and its correlating physiological traits in Drosophila melanogaster. In this study, we conducted a split-family quantitative genetic experiment, in which female adults of Drosophila from 19 isofemale genetic lines were allowed to ingest one of two synthetic diets that differed in protein-to-carbohydrate ratio (P:C = 4:1 or 1:16 with the P+C concentration of 120 g L-1) before they were assayed for starvation time and lipid storage. In all genetic lines, Drosophila flies that had fed carbohydrate-rich diet (P:C=1:16) resisted starvation better and stored more lipids than did those that had fed protein-rich diet (4:1). Importantly, the extent to which both starvation resistance and lipid reserves were affected by dietary P:C ratio varied greatly among different genetic lines of Drosophila, as indicated by significant genotypeby-nutrient interactions for these two traits. When the patterns of the bivariate reaction norm for body lipid and starvation resistance were compared across the genotypes, we found strong evidence for genetic variations in the pattern of energy storage and usage associated with maintaining survival under starvation in Drosophila.

Phytoncides are volatile substances diffused largely from trees to protect themselves against harmful factors. Many people are attracted to forest bathing and the effects of forest bathing involve the effect of protecting human dermal cell against reactive oxygen species (ROS), the activation of immune function and the reduction of stress hormones. Since phytoncides are released to prevent plants from rotting or being eaten by animals, we expect that phytoncides have negative effects on insects. However, there is almost no study to show the effects of phytoncides of Chamaecyparis obtusa on insects so far. Therefore, we examined the effects of phytoncides on insects using fruit fly, Drosophila melanogaster. Our results showed that the exposure to phytoncides scents reduced the lifespan of Drosophila in a dose-dependent manner. Moreover, development rate, locomotion and fecundity of fruit flies were also decreased with phytoncides exposure. In food preference test, fruit flies and house flies showed strong avoidance behavior to the food containing phytoncides in a dose dependent manner. Overall, these results suggest the possibility of phytoncides as human-friendly insect repellent.

Food limitation is the most common environmental challenge faced by animals and the capacity of animals to survive prolonged periods of starvation is linked to their diet and nutritional status. The objective of this study is to investigate the effects of nutrition on starvation resistance in Drosophila melanogaster. Experimental flies were given ad libitum access to artificial diets differing in concentrations and ratios of protein and carbohydrate for 5 days before they were assayed for starvation time, body composition and life-history parameters. Starvation resistance in Drosophila was greatly influenced by the dietary protein:carbohydrate (P:C) ratio, but neither by the caloric content of the diet nor by dietary carbohydrate alone. Starvation resistance was strongest at the lowest P:C ratio and declined with rising P:C ratio. While starving, Drosophila underwent a dramatic transition in the utilization of physiological fuels, switching from the early phase characterized by preferential consumption of non-lipid substrates to the next phase in which they began to mobilize lipids as fuels for enduring starvation. Our results highlight the importance of nutrition as a key factor determining starvation responses of Drosophila.