Drosophila Species Research Articles

Polytene Chromosome Banding Pattern three Drosophila species from Bangladesh

Polytene Chromosome Banding Pattern three Drosophila species from Bangladesh

Ancestral neural circuits potentiate the origin of a female sexual behavior in Drosophila

Courtship interactions are remarkably diverse in form and complexity among species. How neural circuits evolve to encode new behaviors that are functionally integrated into these dynamic social interactions is unknown. Here we report a recently originated female sexual behavior in the island endemic Drosophila species D. santomea, where females signal receptivity to male courtship songs by spreading their wings, which in turn promotes prolonged songs in courting males. Copulation success depends on this female signal and correlates with males’ ability to adjust his singing in such a social feedback loop. Functional comparison of sexual circuitry across species suggests that a pair of descending neurons, which integrates male song stimuli and female internal state to control a conserved female abdominal behavior, drives wing spreading in D. santomea. This co-option occurred through the refinement of a pre-existing, plastic circuit that can be optogenetically activated in an outgroup species. Combined, our results show that the ancestral potential of a socially-tuned key circuit node to engage the wing motor circuit facilitates the expression of a new female behavior in appropriate sensory and motivational contexts. More broadly, our work provides insights into the evolution of social behaviors, particularly female behaviors, and the underlying neural mechanisms.

Females drive postmating reproductive trait evolution across Drosophila species, but not via remating rate.

While traits that contribute to premating sexual interactions are known to be wildly diverse, much less is known about the diversity of postmating (especially female) reproductive traits and the mechanisms shaping this diversity. To assess the rate, pattern, and potential drivers of postmating reproductive trait evolution, we analyzed male and female traits across up to 30 Drosophila species within a phylogenetic comparative framework. In addition to postmating reproductive morphology (e.g., sperm length, reproductive tract length and mass), we also quantified mating behaviors including female remating rate-a common proxy for the strength of postmating sexual selection. We found evidence for strong coevolution between male and female postmating traits (specifically sperm length and sperm storage organ size). However, remating rate was not associated with the rate of evolution or exaggeration of either male or female postmating reproductive morphology, once phylogenetic relatedness was accounted for. We infer that female-mediated and intersexual selection predominantly drive the evolution of our postmating morphological traits, including via divergent male and female interests in controlling paternity. In comparison, remating rate has a complex and likely secondary role in shaping this evolution, in part because this trait can be both a driver and a product of postmating selection.

Classification, biology and entomopathogenic fungi-based management and their mode of action against Drosophila species (Diptera: Drosophilidae): a review.

This review provides a comprehensive analysis of the classification, biology, and management of Drosophila species (Diptera: Drosophilidae) with a focus on entomopathogenic fungi (EPF) as a biocontrol strategy. Drosophila species, particularly Drosophila suzukii, and Drosophila melanogaster have emerged as significant pests in various agricultural systems, causing extensive damage to fruit crops. Understanding their taxonomic classification and biological traits is crucial for developing effective management strategies. This review delves into the life cycle, behavior, and ecological interactions of Drosophila species, highlighting the challenges posed by their rapid reproduction and adaptability. The review further explores the potential of EPF as an eco-friendly alternative to chemical pesticides. The mode of action of EPF against Drosophila species is examined, including spore adhesion, germination, and penetration of the insect cuticle, leading to host death. Factors influencing the efficacy of EPF, such as environmental conditions, fungal virulence, and host specificity, are discussed in detail. By synthesizing current research, this review aims to provide valuable insights into the application of EPF and to identify future research directions for enhancing the effectiveness of EPF-based control measures against Drosophila species.

Adhesive and mechanical properties of the glue produced by 25 Drosophila species

Drosophila glue, a bioadhesive produced by fly larvae to attach themselves to a substrate for several days, has recently gained attention for its peculiar adhesive and mechanical properties. Although Drosophila glue production was described more than 50 years ago, a general survey of the adhesive and mechanical properties of this proteinaceous gel across Drosophila species is lacking. To measure adhesion, we present here a protocol that is robust to variations in protocol parameters, pupal age and calculation methods. We find that the glue, which covers the entire pupal surface, increases the animal rigidity and plasticity when bound to a glass slide. Our survey of pupal adhesion in 25 Drosophilidae species reveals a wide range of phenotypes, from species that produce no or little glue and adhere little, to species that produce high amounts of glue and adhere strongly. One species, D. hydei, stands out from the rest and emerges as a promising model for the development of future bioadhesives, as it has the highest detachment force per glue area and produces relatively large amounts of glue relative to its size. We also observe that species that invest more in glue tend to live in more windy and less rainy climates, suggesting that differences in pupal adhesion properties across species are shaped by ecological factors. Our present survey provides a basis for future biomimetic studies based on Drosophila glue.

Transposable elements in Drosophila montana from harsh cold environments

BackgroundSubstantial discoveries during the past century have revealed that transposable elements (TEs) can play a crucial role in genome evolution by affecting gene expression and inducing genetic rearrangements, among other molecular and structural effects. Yet, our knowledge on the role of TEs in adaptation to extreme climates is still at its infancy. The availability of long-read sequencing has opened up the possibility to identify and study potential functional effects of TEs with higher precision. In this work, we used Drosophila montana as a model for cold-adapted organisms to study the association between TEs and adaptation to harsh climates.ResultsUsing the PacBio long-read sequencing technique, we de novo identified and manually curated TE sequences in five Drosophila montana genomes from eco-geographically distinct populations. We identified 489 new TE consensus sequences which represented 92% of the total TE consensus in D. montana. Overall, 11–13% of the D. montana genome is occupied by TEs, which as expected are non-randomly distributed across the genome. We identified five potentially active TE families, most of them from the retrotransposon class of TEs. Additionally, we found TEs present in the five analyzed genomes that were located nearby previously identified cold tolerant genes. Some of these TEs contain promoter elements and transcription binding sites. Finally, we detected TEs nearby fixed and polymorphic inversion breakpoints.ConclusionsOur research revealed a significant number of newly identified TE consensus sequences in the genome of D. montana, suggesting that non-model species should be studied to get a comprehensive view of the TE repertoire in Drosophila species and beyond. Genome annotations with the new D. montana library allowed us to identify TEs located nearby cold tolerant genes, and present at high population frequencies, that contain regulatory regions and are thus good candidates to play a role in D. montana cold stress response. Finally, our annotations also allow us to identify for the first time TEs present in the breakpoints of three D. montana inversions.

The Ortholog Receptor Or67d in Drosophila Bipectinata is able to Detect Two Different Pheromones

Sex pheromones play a crucial role in species recognition and reproductive isolation. Despite being largely species-specific in drosophilids, the mechanisms underlying pheromone detection, production, and their influence on mating behavior remain poorly understood. Here, we compare the chemical profiles of Drosophila bipectinata and D. melanogaster, the mating behaviors in both species, as well as the tuning properties of Or67d receptors, which are expressed by neurons in antennal trichoid sensilla at1. Through single sensillum recordings, we demonstrate that the D. bipectinata Or67d-ortholog exhibits similar sensitivity to cis-vaccenyl acetate (cVA) as compared to D. melanogaster but in addition also responds uniquely to (Z)-11-eicosen-1-yl-acetate (Z11-20:Ac), a compound exclusively produced by D. bipectinata males. Through courtship behavior assays we found that, surprisingly, perfuming the flies with Z11-20:Ac did not reveal any aphrodisiacal or anti-aphrodisiacal effects in mating assays. The behavioral relevance of at1 neuron channels in D. bipectinata compared to D. melanogaster seems to be restricted to its formerly shown function as an aggregation pheromone. Moreover, the non-specific compound cVA affected copulation negatively in D. bipectinata and could potentially act as a premating isolation barrier. As both ligands of Or67d seem to govern different behaviors in D. bipectinata, additional neurons detecting at least one of those compounds might be involved. These results underscore the complexity of chemical signaling in species recognition and raise intriguing questions about the evolutionary implications of pheromone detection pathways in Drosophila species.

Divergent visual ecology of Drosophila species drives object-tracking strategies matched to landscape sparsity

Maintaining stable gaze while tracking moving objects is commonplace across animal taxa, yet how diverse ecological needs impact these processes is poorly understood. During flight, the fruit-eating fly Drosophila melanogaster maintains course by making smooth steering adjustments to fixate the image of the distant visual background on the retina, while executing body saccades to investigate nearby objects such as food sources. Cactophilic Drosophila mojavensis live where there is no canopy; rather, the flora forming visual "background" and "objects" are one and the same. We tested whether D.mojavensis have adapted their flight control strategies for a visually sparse landscape. We used a magnetic tether that allows free movement in the yaw axis. In response to a textured bar moving across a similarly textured stationary background, D.melanogaster fixates the background, thereby stabilizing gaze while integrating bar dynamics to trigger tracking saccades. By contrast, two mojavensis subspecies in the repleta subgroup and one species in the melanogaster subgroup steer to smoothly fixate the bar, seemingly ignoring the stationary surround. Desert flies execute frequent bar-tracking saccades, but theirs are triggered when rotational velocity lags the bar. Thus, D.melanogaster, which lives in visually cluttered cosmopolitan habitats, leverages the optical disparities between nearby objects and distant foliage for a hybrid control strategy: "ground-fixate, object-saccade." Flies in distant phylogenetic subgroups with similar visual ecology use a "fixate-and-saccade" strategy, which would be adaptive in a visually sparse environment where individual landscape features are both approached and used to maintain a straight course.

FlaHMM: unistrand flamenco-like piRNA cluster prediction in Drosophila species using hidden Markov models.

PIWI-interacting RNAs (piRNAs) are a class of small non-coding RNAs that are essential for transposon control in animal gonads. In Drosophila ovarian somatic cells, piRNAs are transcribed from large genomic regions called piRNA clusters, which are enriched for transposon fragments and act as a memory of past invasions. Despite being widely present across Drosophila species, somatic piRNA clusters are difficult to identify and study due to their lack of sequence conservation and limited synteny. Current identification methods rely on either extensive manual curation or availability of high-throughput small RNA sequencing data, limiting large-scale comparative studies. We now present FlaHMM, a hidden Markov model developed to automate genomic annotation of flamenco-like unistrand piRNA clusters in Drosophila species, requiring only a genome assembly and transposon annotations. FlaHMM uses transposable element content across 5- or 10-kb bins, which can be calculated from genome sequence alone, and is thus able to detect candidate piRNA clusters without the need to obtain flies and experimentally perform small RNA sequencing. We show that FlaHMM performs on par with piRNA-guided or manual methods, and thus provides a scalable and efficient approach to piRNA cluster annotation in new genome assemblies. FlaHMM is freely available at https://github.com/Hannon-lab/FlaHMM under an MIT licence.

Temperature-mediated dynamics: Unravelling the impact of temperature on cuticular hydrocarbon profiles, mating behaviour, and life history traits in three Drosophila species

In a world grappling with climate change, understanding the enduring impact of changes in temperatures on insect adult traits is crucial. It is proposed that cold- and warm-adapted species exhibit specialized behavioural and physiological responses to their respective temperature ranges. In contrast, generalist species maintain more stable metabolic and developmental rates across a broader range of temperatures, reflecting their ability to exploit diverse thermal niches. Here, we explored this intricate response to temperature exposure in three Drosophila species: Drosophila ezoana originating in Arctic regions, D. novamexicana in arid, hot environments, and in the cosmopolitan species D. virilis. Rearing these flies at 15, 20, 25, and 30 °C revealed striking variations in their cuticular hydrocarbon (CHC) profiles, known to mediate mate recognition and prevent water loss in insects. The cold-adapted D. ezoana consistently exhibited reduced CHC levels with increasing temperatures, while the warm-adapted D. novamexicana and the cosmopolitan D. virilis displayed more nuanced responses. Additionally, we observed a significant influence of rearing temperature on the mating behaviour of these flies, where those reared at the extreme temperatures, 15 and 30 °C, exhibiting reduced mating success. Consequently, this led to a decrease in the production of adult offspring. Also, these adult offspring underwent notable alterations in life history traits, reaching adulthood more rapidly at 25 and 30 °C but with lower weight and reduced longevity. Furthermore, among these offspring, those produced by the cold-adapted D. ezoana were more vulnerable to desiccation and starvation than those from the warm-adapted D. novamexicana and the cosmopolitan D. virilis. In summary, our research demonstrates that Drosophila species from diverse ecological regions exhibit distinct responses to temperature changes, as evidenced by variations in CHC profiles, mating behaviours, fertility, and life history traits. This provides valuable insights into how environmental conditions shape the biology and ecology of insects.

Protocol for dissection of Drosophila abdomens for fluorescent imaging

Fluorescent imaging in Drosophila species is indispensable for investigating dynamic biological processes and visualizing gene expression patterns with cellular precision. This technique leverages the transparency of Drosophila larvae and pupae, combined with advanced microscopy, to enable real-time observation of developmental events such as morphogenesis and organogenesis. Genetically encoded fluorescent proteins and dyes allow specific labeling of cells and proteins, facilitating detailed studies of spatial and temporal dynamics within intact tissues. Techniques like confocal and two-photon microscopy provide high resolution and depth penetration, essential for 3D reconstruction and quantitative analysis of complex biological structures. Fluorescent imaging in Drosophila supports disease modeling, drug screening, and therapeutic exploration, bridging insights from basic biology to potential clinical applications. It is therefore necessary to develop imaging techniques and protocols that accurately capture and profile gene expression patterns in a wide range of Drosophila tissues. In this study, we present a detailed protocol for preparing and imaging transgenic Drosophila abdomen, which will enable researchers investigate gene expression patterns underlying fundamental biological processes in the abdomen.

Response of fruit fly (Drosophila pseudoobscura) to diet manipulation of nutrient density

AbstractCaloric intake can greatly affect many aspects of an organism's life. A deficiency of calories can lead to stress resulting in decreased fecundity, insufficient calories to maintain tissues and increased lifespan. Conversely, increasing caloric density increases fecundity and decreases lifespan. Despite decades of work exploring food quality and quantity on physiology in the model species Drosophila melanogaster Meigan 1830 (Diptera: Drosophilidae) and the melanogaster group in general, relatively little work explores the physiological responses to diet manipulation in other Drosophila species, like the obscura species group. Here, we looked at the effects of five different caloric densities (0.5×, 0.75×, 1.0×, 1.5× and 3.0×) on food intake, body weight, body fat, fecundity and longevity in D. pseudoobscura Frolova & Astaurov, 1929 (Diptera: Drosophilidae). Comparing longevity and fecundity across diets, we found that heavy caloric concentration (3.0×) decreases lifespan and that calorie restriction (0.5× and 0.75×) led to significant decreases in fecundity and body weight. However, calorie concentration did not significantly increase D. pseudoobscura body fat. By expanding our understanding of the physiological responses to diet stress to D. pseudoobscura, we establish the framework for comparative work across Drosophila species. With this information, we can then identify which physiological responses to diet manipulation might be most conserved and comparable across species.

Evolution of sex-biased genes in Drosophila species with neo-sex chromosomes: Potential contribution to reducing the sexual conflict.

An advantage of sex chromosomes may be the potential to reduce sexual conflict because they provide a basis for selection to operate separately on females and males. However, evaluating the relationship between sex chromosomes and sexual conflict is challenging owing to the difficulty in measuring sexual conflict and substantial divergence between species with and without sex chromosomes. We therefore examined sex-biased gene expression as a proxy for sexual conflict in three sets of Drosophila species with and without young sex chromosomes, the so-called neo-sex chromosomes. In all sets, we detected more sex-biased genes in the species with neo-sex chromosomes than in the species without neo-sex chromosomes in larvae, pupae, and adult somatic tissues but not in gonads. In particular, many unbiased genes became either female- or male-biased after linkage to the neo-sex chromosomes in larvae, despite the low sexual dimorphism. For example, genes involved in metabolism, a key determinant for the rate of development in many animals, were enriched in the genes that acquired sex-biased expression on the neo-sex chromosomes at the larval stage. These genes may be targets of sexually antagonistic selection (i.e., large size and rapid development are selected for in females but selected against in males). These results indicate that acquiring neo-sex chromosomes may have contributed to a reduction in sexual conflict, particularly at the larval stage, in Drosophila..

Characterization of pre-diapause phase in the northern Drosophila species D. ezoana.

Drosophila ezoana is a virilis group Drosophila species inhabiting northern latitudes. The flies enter adult reproductive diapause to survive winter upon exposure to short photoperiod conditions (short-day) over several consecutive days. Insect pre-diapause phase - the duration between the beginning of exposure to short days and expression of diapause is thought to be comprised of two distinct phases - (a) photoperiodic time measurement that detects short-days, followed by (b) physiological events leading to the expression of diapause phenotype. A short-day dependent segment of the pre-diapause phase thus approximates the process of photoperiodic time measurement. Continuous darkness has been found to be a neutral condition with respect to diapause regulation in many insect species. The effect of variable number of short-days followed by continuous darkness on diapause incidence thus allows identification of short-day dependent segment of pre-diapause phase thereby mapping the process of photo-periodic time measurement. Although, few weeks of exposure to short-days in adult stage is known to be sufficient for the expression of diapause in D. ezoana, the number of short days required for the completion of photo-periodic time measurement has never been systematically analysed. Our experiments show that continuous darkness is a neutral condition for diapause regulation also in D. ezoana. We utilized the neutral nature of continuous darkness to map the process of photoperiodic time measurement in the D. ezoana strain 124OJ8 which showed that integration of short-day photic cues over the first 10 days of pre-diapause phase is essential for diapause induction.

No evidence for the melanin desiccation hypothesis in a larval Lepidopteran

Water regulation is an important physiological challenge for insects due to their small body sizes and large surface area to volume ratios. Adaptations for decreasing cuticular water loss, the largest avenue of loss, are especially important. The melanin desiccation hypothesis states that melanin molecules in the cuticle may help prevent water loss, thus offering protection from desiccation. This hypothesis has much empirical support in Drosophila species, but remains mostly untested in other taxa, including Lepidoptera. Because melanin has many other important functions in insects, its potential role in desiccation prevention is not always clear. In this study we investigated the role of melanin in desiccation prevention in the white-lined Sphinx moth, Hyles lineata (Lepidoptera, Sphingidae), which shows high plasticity in the degree of melanin pigmentation during the late larval instars. We took advantage of this plasticity and used density treatments to induce a wide range of cuticular melanization; solitary conditions induced low melanin pigmentation while crowded conditions induced high melanin pigmentation. We tested whether more melanic larvae from the crowded treatment were better protected from desiccation in three relevant responses: i) total water loss over a desiccation period, ii) change in hemolymph osmolality over a desiccation period, and iii) evaporation rate of water through the cuticle. We did not find support for the melanin desiccation hypothesis in this species. Although treatment influenced total water loss, this effect did not occur via degree of melanization. Interestingly, this implies that crowding, which was used to induce high melanin phenotypes, may have other physiological effects that influence water regulation. There were no differences between treatments in cuticular evaporative water loss or change in hemolymph osmolality. However, we conclude that osmolality may not sufficiently reflect water loss in this case. This study emphasizes the context dependency of melanin’s role in desiccation prevention and the importance of considering how it may vary across taxa. In lepidopteran larvae that are constantly feeding phytophagous insects with soft cuticles, melanin may not be necessary for preventing cuticular water loss.

Altered circadian rhythm, sleep, and rhodopsin 7–dependent shade preference during diapause in Drosophila melanogaster

To survive adverse environments, many animals enter a dormant state such as hibernation, dauer, or diapause. Various Drosophila species undergo adult reproductive diapause in response to cool temperatures and/or short day-length. While flies are less active during diapause, it is unclear how adverse environmental conditions affect circadian rhythms and sleep. Here we show that in diapause-inducing cool temperatures, Drosophila melanogaster exhibit altered circadian activity profiles, including severely reduced morning activity and an advanced evening activity peak. Consequently, the flies have a single activity peak at a time similar to when nondiapausing flies take a siesta. Temperatures ≤15 °C, rather than photoperiod, primarily drive this behavior. At cool temperatures, flies rapidly enter a deep-sleep state that lacks the sleep cycles of flies at higher temperatures and require high levels of stimulation for arousal. Furthermore, we show that at 25 °C, flies prefer to siesta in the shade, a preference that is virtually eliminated at 10 °C. Resting in the shade is driven by an aversion to blue light that is sensed by Rhodopsin 7 outside of the eyes. Flies at 10 °C show neuronal markers of elevated sleep pressure, including increased expression of Bruchpilot and elevated Ca2+ in the R5 ellipsoid body neurons. Therefore, sleep pressure might overcome blue light aversion. Thus, at the same temperatures that cause reproductive arrest, preserve germline stem cells, and extend lifespan, D. melanogaster are prone to deep sleep and exhibit dramatically altered, yet rhythmic, daily activity patterns.

Divergent evolution of sleep in Drosophila species

Living organisms synchronize their biological activities with the earth’s rotation through the circadian clock, a molecular mechanism that regulates biology and behavior daily. This synchronization factually maximizes positive activities (e.g., social interactions, feeding) during safe periods, and minimizes exposure to dangers (e.g., predation, darkness) typically at night. Beyond basic circadian regulation, some behaviors like sleep have an additional layer of homeostatic control, ensuring those essential activities are fulfilled. While sleep is predominantly governed by the circadian clock, a secondary homeostatic regulator, though not well-understood, ensures adherence to necessary sleep amounts and hints at a fundamental biological function of sleep beyond simple energy conservation and safety. Here we explore sleep regulation across seven Drosophila species with diverse ecological niches, revealing that while circadian-driven sleep aspects are consistent, homeostatic regulation varies significantly. The findings suggest that in Drosophilids, sleep evolved primarily for circadian purposes. The more complex, homeostatically regulated functions of sleep appear to have evolved independently in a species-specific manner, and are not universally conserved. This laboratory model may reproduce and recapitulate primordial sleep evolution.

Microsatellite polymorphism based genetic variation in two sympatric species of Drosophila: D.bipectinata and D. malerkotliana

Drosophila bipectinata and D. malerkotliana are found to be distributed all along the Oriental-Australian continents and have common phylogenetic descent. The genetic divergence of these two species have been studied by using genetic markers like chromosome inversions and allozyme variants to establish the extent of variation they have experienced since their speciation. In this study, microsatellite variations have been considered to observe the genetic differentiation among the seven natural populations of D. bipectinata and six populations of D. malerkotliana. Results pertaining to both the species indicate that the level of average heterozygosity increases from north to south. Distinct genetic differentiation has been observed between north and south populations of both the species giving an indication that north and south populations have undergone genetic structuring due to absence of gene flow between them and varying environmental conditions. Based on the frequencies of microsatellite variants of the two species, we have computed the level of variation between these two species. The analysis in this regard shows an almost similar trend of genetic variation being experienced by these two species which could be a kind of parallel evolution occurring due to similar ecological niches occupied by them and similar impact of evolutionary forces.

Gene Protein Sequence Evolution Can Predict the Rapid Divergence of Ovariole Numbers in the Drosophila melanogaster Subgroup.

Ovaries play key roles in fitness and evolution: they are essential female reproductive structures that develop and house the eggs in sexually reproducing animals. In Drosophila, the mature ovary contains multiple tubular egg-producing structures known as ovarioles. Ovarioles arise from somatic cellular structures in the larval ovary called terminal filaments (TFs), formed by TF cells and subsequently enclosed by sheath (SH) cells. As in many other insects, ovariole number per female varies extensively in Drosophila. At present, however, there is a striking gap of information on genetic mechanisms and evolutionary forces that shape the well-documented rapid interspecies divergence of ovariole numbers. To address this gap, here we studied genes associated with Drosophila melanogaster ovariole number or functions based on recent experimental and transcriptional datasets from larval ovaries, including TFs and SH cells, and assessed their rates and patterns of molecular evolution in five closely related species of the melanogaster subgroup that exhibit species-specific differences in ovariole numbers. From comprehensive analyses of protein sequence evolution (dN/dS), branch-site positive selection, expression specificity (tau), and phylogenetic regressions (phylogenetic generalized least squares), we report evidence of 42 genes that showed signs of playing roles in the genetic basis of interspecies evolutionary change of Drosophila ovariole number. These included the signaling genes upd2 and Ilp5 and extracellular matrix genes vkg and Col4a1, whose dN/dS predicted ovariole numbers among species. Together, we propose a model whereby a set of ovariole-involved gene proteins have an enhanced evolvability, including adaptive evolution, facilitating rapid shifts in ovariole number among Drosophila species.

Evolution of connectivity architecture in the Drosophila mushroom body

Brain evolution has primarily been studied at the macroscopic level by comparing the relative size of homologous brain centers between species. How neuronal circuits change at the cellular level over evolutionary time remains largely unanswered. Here, using a phylogenetically informed framework, we compare the olfactory circuits of three closely related Drosophila species that differ in their chemical ecology: the generalists Drosophila melanogaster and Drosophila simulans and Drosophila sechellia that specializes on ripe noni fruit. We examine a central part of the olfactory circuit that, to our knowledge, has not been investigated in these species—the connections between projection neurons and the Kenyon cells of the mushroom body—and identify species-specific connectivity patterns. We found that neurons encoding food odors connect more frequently with Kenyon cells, giving rise to species-specific biases in connectivity. These species-specific connectivity differences reflect two distinct neuronal phenotypes: in the number of projection neurons or in the number of presynaptic boutons formed by individual projection neurons. Finally, behavioral analyses suggest that such increased connectivity enhances learning performance in an associative task. Our study shows how fine-grained aspects of connectivity architecture in an associative brain center can change during evolution to reflect the chemical ecology of a species.