Chemical and Electrophysiological Characterisation of Headspace Volatiles from Yeasts Attractive to Drosophila suzukii.

Chemical control of Drosophila suzukii (Diptera: Drosophilidae) based on the use of insecticides is particularly challenging as the insect attacks ripening fruits shortly before harvest. An alternative strategy may rely on the use of yeasts as phagostimulants and baits, applied on canopy as attract-and-kill formulations. The aim of this research was to identify the most attractive among six yeast species for D. suzukii: Saccharomyces cerevisiae, Hanseniaspora uvarum, Clavispora santaluciae, Saccharomycopsis vini, Issatchenkia terricola, and Metschnikowia pulcherrima. The volatile profile of C. santaluciae was described for the first time. Behavioural experiments identified H. uvarum and S. vini as the most attractive yeasts. The characterization of yeast headspace volatiles using direct headspace (DHS) and solid-phase microextraction (SPME) revealed several strain-specific compounds. With DHS injection, 19 volatiles were characterised, while SPME revealed 71 compounds constituting the yeast headspace. Both analyses revealed terpenoids including ß-ocimene, citronellol, (Z)-geraniol (nerol), and geranial as distinct constituents of S. vini. H. uvarum and S. vini were further investigated using closed-loop stripping analysis (CSLA) and electroantennography. Out of 14 compounds quantified by CSLA, ethyl acetate, isoamyl acetate, ß-myrcene, benzaldehyde and linalool were detected by D. suzukii antennae and might generate the strong attractiveness of S. vini and H. uvarum. Our results highlight a strong attraction of D. suzukii to various yeasts associated with both the flies and their habitat and demonstrate how different sampling methods can impact the results of volatile compound characterization. It remains to be demonstrated whether the distinct attraction is based on special adaptations to certain yeasts and to what extent the metabolites causing attraction are interchangeable.

Microbiota-mediated competition between Drosophila species.

BACKGROUND: The influence of microbiota in ecological interactions, and in particular competition, is poorly known. We studied competition between two insect species, the invasive pest Drosophila suzukii and the model Drosophila melanogaster, whose larval ecological niches overlap in ripe, but not rotten, fruit. RESULTS: We discovered D. suzukii females prevent costly interspecific larval competition by avoiding oviposition on substrates previously visited by D. melanogaster. More precisely, D. melanogaster association with gut bacteria of the genus Lactobacillus triggered D. suzukii avoidance. However, D. suzukii avoidance behavior is condition-dependent, and D. suzukii females that themselves carry D. melanogaster bacteria stop avoiding sites visited by D. melanogaster. The adaptive significance of avoiding cues from the competitor's microbiota was revealed by experimentally reproducing in-fruit larval competition: reduced survival of D. suzukii larvae only occurred if the competitor had its normal microbiota. CONCLUSIONS: This study establishes microbiotas as potent mediators of interspecific competition and reveals a central role for context-dependent behaviors under bacterial influence. Video Abstract.

Political deafness may impede transition to biological control.

Biological control has developed into a realistic alternative to replace chemical pesticides. A long-awaited paradigm shift is now adopted by the European Commission through a proposed new Regulation on sustainable use of plant protection products. Unfortunately, the scientific framework underpinning biocontrol is seriously neglected, impeding transition to sustainable plant production.

Antennal and maxillary palp morphology, and sensillar equipment, of the spruce bark beetle predators, Medetera signaticornis and Medetera infumata (Diptera: Dolichopodidae).

Many long-legged Medetera flies are natural enemies of bark beetle pests, which they detect using olfactory cues, likely through olfactory sensilla on the antennae and maxillary palps. Morphological characterisation of olfactory sensilla among insects can provide a basis for future taxonomic, phylogenetic or electrophysiological studies. Scanning electron microscopy was used to describe the morphology of olfactory organs and sensillar equipment of Medetera signaticornis and M. infumata. Three different olfactory sensillum types were found in both fly species, sensilla trichodea, s. basiconica and grooved pegs. Based on size and wall structure, s. trichodea and s. basiconica were categorised into different subtypes. Sharp-tipped curved s. trichodea, and small, large and thin s. basiconica were found on the antennal postpedicel of M. signaticornis adults, while grooved s. basiconica were found in M. infumata. The density of sharp-tipped long s. trichodea was significantly higher in males compared to females, and in M. signaticornis compared to M. infumata. Long-grooved s. basiconica were found grouped in a small pit on the maxillary palps of both species. Comparison of our results with the limited available ecological data suggests that differences in numbers of specific sensillum types may reflect adaptations related to olfactory-driven behaviours such as host seeking.

Odors Attracting the Long-Legged Predator Medetera signaticornis Loew to Ips typographus L. Infested Norway Spruce Trees.

Predatory long-legged flies of the genus Medetera are important, but currently understudied, natural enemies of Scolytinae bark beetles such as Ips typographus. Medetera flies lay eggs on beetle-infested trees, where the developing larvae find their prey, but the chemical cues used by Medetera to locate infested trees are currently unknown. To identify odors attracting Medetera signaticornis, a species in Europe, headspace samples were collected at several time-points through different stages of I. typographus attacks on logs of Norway spruce (Picea abies). The headspace samples were analyzed using combined gas chromatography and mass spectrometry (GC-MS), and gas chromatography coupled with electroantennographic detection (GC-EAD) to determine compounds that stimulate M. signaticornis antennae. Antennae of M. signaticornis males and females were found to detect (-)-cis-verbenol, ( +)-trans-verbenol and myrtenol, which are known to be produced by bark beetles. Antennal responses were also observed for verbenene, isoterpinolene, α-pinene oxide, camphor, pinocamphone, terpinene-4-ol, myrtenal, borneol, α-terpineol, geranyl acetone, and verbenone, which are primarily produced by microorganisms, and α-pinene, α-fenchene, ß-pinene, camphene, 3-carene, limonene, γ-terpinene, and terpinolene, known spruce tree compounds. In field experiments testing two synthetic blends containing 18 antennal active and two additional compounds 2-methyl-3-buten-2-ol and ipsdienol we observed significant attraction of M. signaticornis within 24 h. These attractive blends can form the basis for development of Medetera monitoring lures for use in future forest and pest management.

Comparative transcriptomic assessment of the chemosensory receptor repertoire of Drosophila suzukii adult and larval olfactory organs.

The spotted wing Drosophila, Drosophila suzukii, has emerged within the past decade as an invasive species on a global scale, and is one of the most economically important pests in fruit and berry production in Europe and North America. Insect ecology, to a strong degree, depends on the chemosensory modalities of smell and taste. Extensive research on the sensory receptors of the olfactory and gustatory systems in Drosophila melanogaster provide an excellent frame of reference to better understand the fundamentals of the chemosensory systems of D. suzukii. This knowledge may enhance the development of semiochemicals for sustainable management of D. suzukii, which is urgently needed. Here, using a transcriptomic approach we report the chemosensory receptor expression profiles in D. suzukii female and male antennae, and for the first time, in larval heads including the dorsal organ that houses larval olfactory sensory neurons. In D. suzukii adults, we generally observed a lack of sexually dimorphic expression levels in male and female antennae. While there was generally conservation of antennal expression of odorant and ionotropic receptor orthologues for D. melanogaster and D. suzukii, gustatory receptors showed more distinct species-specific profiles. In larval head tissues, for all three receptor gene families, there was also a greater degree of species-specific gene expression patterns. Analysis of chemosensory receptor repertoires in the pest species, D. suzukii relative to those of the genetic model D. melanogaster enables comparative studies of the chemosensory, physiology, and ecology of D. suzukii.

The human odorant receptor OR10A6 is tuned to the pheromone of the commensal fruit fly Drosophila melanogaster.

All living things speak chemistry. The challenge is to reveal the vocabulary, the odorants that enable communication across phylogenies and to translate them to physiological, behavioral, and ecological function. Olfactory receptors (ORs) interface animals with airborne odorants. Expression in heterologous cells makes it possible to interrogate single ORs and to identify cognate ligands. The cosmopolitan, anthropophilic strain of the vinegar fly Drosophila melanogaster depends on human resources and housing for survival. Curiously, humans sense the pheromone (Z)-4-undecenal (Z4-11Al) released by single fly females. A screening of all human ORs shows that the most highly expressed OR10A6 is tuned to Z4-11Al. Females of an ancestral African fly strain release a blend of Z4-11Al and Z4-9Al that produces a different aroma, which is how we distinguish these fly strains by nose. That flies and humans sense Z4-11Al via dedicated ORs shows how convergent evolution shapes communication channels between vertebrate and invertebrate animals.

Pathogenic fungus uses volatiles to entice male flies into fatal matings with infected female cadavers.

To ensure dispersal, many parasites and pathogens behaviourally manipulate infected hosts. Other pathogens and certain insect-pollinated flowers use sexual mimicry and release deceptive mating signals. However, it is unusual for pathogens to rely on both behavioural host manipulation and sexual mimicry. Here, we show that the host-specific and behaviourally manipulating pathogenic fungus, Entomophthora muscae, generates a chemical blend of volatile sesquiterpenes and alters the profile of natural host cuticular hydrocarbons in infected female housefly (Musca domestica) cadavers. Healthy male houseflies respond to the fungal compounds and are enticed into mating with female cadavers. This is advantageous for the fungus as close proximity between host individuals leads to an increased probability of infection. The fungus exploits the willingness of male flies to mate and benefits from altering the behaviour of uninfected male host flies. The altered cuticular hydrocarbons and emitted volatiles thus underlie the evolution of an extended phenotypic trait.

Hanseniaspora uvarum Attracts Drosophila suzukii (Diptera: Drosophilidae) With High Specificity.

Since the early phase of the intercontinental dispersal of Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), fermentation baits have been used for monitoring. Self-made lures and commercial products are often based on wine and vinegar. From an ecological perspective, the formulation of these baits is expected to target especially vinegar flies associated with overripe fruit, such as Drosophila melanogaster (Meigen) (Diptera: Drosophilidae). Hanseniaspora uvarum (Niehaus) (Ascomycota: Saccharomyceta) is a yeast closely associated with D. suzukii and fruit, and furthermore attractive to the flies. Based on this relation, H. uvarum might represent a suitable substrate for the development of lures that are more specific than vinegar and wine. In the field, we therefore, compared H. uvarum to a commercial bait that was based on vinegar and wine with respect to the number of trapped D. suzukii relative to other drosophilids and arthropods. Trap captures were higher with the commercial bait but specificity for D. suzukii was greater with H. uvarum. Moreover, H. uvarum headspace extracts, as well as a synthetic blend of H. uvarum volatiles, were assayed for attraction of D suzukii in a wind tunnel and in the field. Headspace extracts and the synthetic blend induced strong upwind flight in the wind tunnel and confirmed attraction to H. uvarum volatiles. Furthermore, baited with H. uvarum headspace extract and a drowning solution of aqueous acetic acid and ethanol, 74% of field captured arthropods were D. suzukii. Our findings suggest that synthetic yeast headspace formulations might advance the development of more selective monitoring traps with reduced by-catch.

The female sex pheromone (Z)-4-undecenal mediates flight attraction and courtship in Drosophila melanogaster.

Specific mate communication and recognition underlies reproduction and hence speciation. Our study provides new insights in Drosophila melanogaster premating olfactory communication. Mate communication evolves during adaptation to ecological niches and makes use of social signals and habitat cues. Female-produced, species-specific volatile pheromone (Z)-4-undecenal (Z4-11Al) and male pheromone (Z)-11-octadecenyl acetate (cVA) interact with food odour in a sex-specific manner. Furthermore, Z4-11Al, which mediates upwind flight attraction in both sexes, also elicits courtship in experienced males. Two isoforms of the olfactory receptor Or69a are co-expressed in the same olfactory sensory neurons. Z4-11Al is perceived via Or69aB, while the food odorant (R)-linalool is a main ligand for the other variant, Or69aA. However, only Z4-11Al mediates courtship in experienced males, not (R)-linalool. Behavioural discrimination is reflected by calcium imaging of the antennal lobe, showing distinct glomerular activation patterns by these two compounds. Male sex pheromone cVA is known to affect male and female courtship at close range, but does not elicit upwind flight attraction as a single compound, in contrast to Z4-11Al. A blend of the food odour vinegar and cVA attracted females, while a blend of vinegar and female pheromone Z4-11Al attracted males, instead. Sex-specific upwind flight attraction to blends of food volatiles and male and female pheromone, respectively, adds a new element to Drosophila olfactory premating communication and is an unambiguous paradigm for identifying the behaviourally active components, towards a more complete concept of food-pheromone odour objects.

Behavioral manipulation of Drosophila suzukii for pest control: high attraction to yeast enhances insecticide efficacy when applied on leaves.

BACKGROUND: The invasive pest, Drosophila suzukii attacks fresh soft-skinned fruit. Broad-spectrum insecticides are implemented for control but there is a need to reduce environmental risks and insecticide residues on fruits. Hanseniaspora uvarum is a yeast frequently found on ripe fruits and associated with D. suzukii. We aim to exploit the ecological association and attraction of D. suzukii to H. uvarum by developing an attract-and-kill strategy, with spray-application on canopy but not fruit. We therefore investigated D. suzukii attraction, egg-laying and mortality when exposed to insecticidal yeast-based formulations. RESULTS: Hanseniaspora uvarum strongly attracted D. suzukii when applied on leaves of grapevine, Vitis vinifera. Notably, this attractiveness was competitive to ripe grape berries that were susceptible to D. suzukii infestation. Moreover, adding H. uvarum enhanced the efficacy of insecticidal formulations against D. suzukii. Flies exposed to leaves treated with yeast-insecticide formulations showed higher mortality and laid a lower number of eggs compared to flies exposed to insecticide alone. In a wind tunnel, all treatments containing H. uvarum alone or in combination with insecticides, caused similar upwind flight and landing at the odor source, which provides evidence that the addition of insecticide did not reduce D. suzukii attraction to yeast. CONCLUSION: Hanseniaspora uvarum can be used to manipulate the behavior of D. suzukii by attracting flies to insecticide formulations. Yeast attraction is competitive to grape berries and improves insecticide effectiveness, suggesting that sprays covering canopy only, could reduce residues on fruit without compromising management efficacy.

Field and greenhouse application of an attract-and-kill formulation based on the yeast Hanseniaspora uvarum and the insecticide spinosad to control Drosophila suzukii in grapes.

BACKGROUND: The invasive insect Drosophila suzukii (Matsumura) is an important pest of several red grape varieties. The yeast Hanseniaspora uvarum (Niehaus), which is associated with D. suzukii, strongly attracts flies and stimulates them to feed on yeast-laden food. In the present study, a formulation based on H. uvarum culture with spinosad insecticide was applied to the foliage of vineyards and control of D. suzukii was compared to applying spinosad to the whole plant. After successful H. uvarum and insecticide application in the vineyard, we tested additional H. uvarum-based formulations with spinosad in a greenhouse to determine their capacity to control D. suzukii. RESULTS: Application of the H. uvarum-spinosad formulation at 36.4 g of spinosad per hectare reduced the D. suzukii field infestation at the same rate as applying 120 g of spinosad per hectare and prevented spinosad residues on grapes. Leaves treated with H. uvarum and spinosad in the field and transferred to a laboratory assay caused high mortality to flies and reduced the number of eggs laid on fruits. Formulations with spinosad applied in the greenhouse showed that both H. uvarum culture and the yeast cell-free supernatant of a centrifuged culture increased fly mortality and reduced the number of eggs laid compared to the unsprayed control. CONCLUSION: In comparison to typical spinosad spray applications, the use of H. uvarum in combination with spinosad as an attract-and-kill formulation against D. suzukii reduces pesticide residues on the fruits by targeting the treatment to the canopy and decreasing the amount of insecticide per hectare without compromising control efficacy.

Stage-specific expression of an odorant receptor underlies olfactory behavioral plasticity in Spodoptera littoralis larvae.

BACKGROUND: The detection of environmental cues and signals via the sensory system directs behavioral choices in diverse organisms. Insect larvae rely on input from the chemosensory system, mainly olfaction, for locating food sources. In several lepidopteran species, foraging behavior and food preferences change across larval instars; however, the molecular mechanisms underlying such behavioral plasticity during larval development are not fully understood. Here, we hypothesize that expression patterns of odorant receptors (ORs) change during development, as a possible mechanism influencing instar-specific olfactory-guided behavior and food preferences. RESULTS: We investigated the expression patterns of ORs in larvae of the cotton leafworm Spodoptera littoralis between the first and fourth instar and revealed that some of the ORs show instar-specific expression. We functionally characterized one OR expressed in the first instar, SlitOR40, as responding to the plant volatile, ß-caryophyllene and its isomer α-humulene. In agreement with the proposed hypothesis, we showed that first but not fourth instar larvae responded behaviorally to ß-caryophyllene and α-humulene. Moreover, knocking out this odorant receptor via CRISPR-Cas9, we confirmed that instar-specific responses towards its cognate ligands rely on the expression of SlitOR40. CONCLUSION: Our results provide evidence that larvae of S. littoralis change their peripheral olfactory system during development. Furthermore, our data demonstrate an unprecedented instar-specific behavioral plasticity mediated by an OR, and knocking out this OR disrupts larval behavioral plasticity. The ecological relevance of such behavioral plasticity for S. littoralis remains to be elucidated, but our results demonstrate an olfactory mechanism underlying this plasticity in foraging behavior during larval development.

Insect chemical ecology: chemically mediated interactions and novel applications in agriculture.

Insect chemical ecology (ICE) evolved as a discipline concerned with plant-insect interactions, and also with a strong focus on intraspecific pheromone-mediated communication. Progress in this field has rendered a more complete picture of how insects exploit chemical information in their surroundings in order to survive and navigate their world successfully. Simultaneously, this progress has prompted new research questions about the evolution of insect chemosensation and related ecological adaptations, molecular mechanisms that mediate commonly observed behaviors, and the consequences of chemically mediated interactions in different ecosystems. Themed meetings, workshops, and summer schools are ideal platforms for discussing scientific advancements as well as identifying gaps and challenges within the discipline. From the 11th to the 22nd of June 2018, the 11th annual PhD course in ICE was held at the Swedish University of Agricultural Sciences (SLU) Alnarp, Sweden. The course was made up of 35 student participants from 22 nationalities (Fig. 1a) as well as 32 lecturers. Lectures and laboratory demonstrations were supported by literature seminars, and four broad research areas were covered: (1) multitrophic interactions and plant defenses, (2) chemical communication focusing on odor sensing, processing, and behavior, (3) disease vectors, and (4) applied aspects of basic ICE research in agriculture. This particular article contains a summary and brief synthesis of these main emergent themes and discussions from the ICE 2018 course. In addition, we also provide suggestions on teaching the next generation of ICE scientists, especially during unprecedented global situations.

Developmentally regulated volatiles geosmin and 2-methylisoborneol attract a soil arthropod to Streptomyces bacteria promoting spore dispersal.

Volatile compounds emitted by bacteria are often sensed by other organisms as odours, but their ecological roles are poorly understood1,2. Well-known examples are the soil-smelling terpenoids geosmin and 2-methylisoborneol (2-MIB)3,4, which humans and various animals sense at extremely low concentrations5,6. The conservation of geosmin biosynthesis genes among virtually all species of Streptomyces bacteria (and genes for the biosynthesis of 2-MIB in about 50%)7,8, suggests that the volatiles provide a selective advantage for these soil microbes. We show, in the present study, that these volatiles mediate interactions of apparent mutual benefit between streptomycetes and springtails (Collembola). In field experiments, springtails were attracted to odours emitted by Streptomyces colonies. Geosmin and 2-MIB in these odours induce electrophysiological responses in the antennae of the model springtail Folsomia candida, which is also attracted to both compounds. Moreover, the genes for geosmin and 2-MIB synthases are under the direct control of sporulation-specific transcription factors, constraining emission of the odorants to sporulating colonies. F. candida feeds on the Streptomyces colonies and disseminates spores both via faecal pellets and through adherence to its hydrophobic cuticle. The results indicate that geosmin and 2-MIB production is an integral part of the sporulation process, completing the Streptomyces life cycle by facilitating dispersal of spores by soil arthropods.

Leaf mineral content govern microbial community structure in the phyllosphere of spinach (Spinacia oleracea) and rocket (Diplotaxis tenuifolia).

The plant microbiome is an important factor for plant health and productivity. While the impact of nitrogen (N) availability for plant growth and development is well established, its influence on the microbial phyllosphere community structure is unknown. We hypothesize that nitrogen impacts the growth and abundance of several microorganisms on the leaf surface. The bacterial and fungal communities of baby leaf spinach (Spinacia oleracea), and rocket (Diplotaxis tenuifolia) were investigated in a field trial for two years in a commercial setting. Nitrogen fertilizer was tested in four doses (basic nitrogen, basic + suboptimal, basic + commercial, basic + excess) with six replicates in each. Culture-independent (Illumina sequencing) and culture-dependent (viable count and identification of bacterial isolates) community studies were combined with monitoring of plant physiology and site weather conditions. This study found that alpha diversity of bacterial communities decreased in response to increasing nitrogen fertilizer dose, whereas viable counts showed no differences. Correspondingly, fungal communities of the spinach phyllosphere showed a decreasing pattern, whereas the decreasing diversity of fungal communities of rocket was not significant. Plant species and effects of annual variations on microbiome structure were observed for bacterial and fungal communities on both spinach and rocket. This study provides novel insights on the impact of nitrogen fertilizer regime on a nutrient scarce habitat, the phyllosphere.

Functional transcriptome analyses of Drosophila suzukii antennae reveal mating-dependent olfaction plasticity in females.

Insect olfaction modulates basal behaviors and it is often influenced by the physiological condition of each individual such as the reproductive state. Olfactory plasticity can be achieved by modifications at both peripheral and central nervous system levels. Here we performed a genome-wide transcriptomic analysis of the main olfactory organ, the antenna, to investigate how gene expression varies with female mating status in Drosophila suzukii, a destructive and invasive soft fruit pest. We observed a wide mating-induced up-regulation of chemosensory-related genes in females, especially odorant receptor (Or) genes. We then used a candidate gene approach to define the comprehensive dataset of antenna-expressed chemosensory receptors and binding proteins, which showed many similarities with Drosophila melanogaster. Candidate gene approach was also used to finely quantify differential expression at Or isoform level, suggesting post-mating transcriptional modulation of genes involved in the peripheral olfactory system. We identified 27 up-regulated Or transcripts encoded by 25 genes, seven of them were duplications specific to D. suzukii lineage. Post-mating olfactory modulation was further supported by electroantennogram recordings that showed a differential response according to mating status to one out of eight odors tested (isoamyl-acetate). Our study characterizes the transcriptional mechanisms driven by mating in D. suzukii female antennae. Understanding the role of genes differentially expressed in virgin or mated females will be crucial to better understand host finding and the crop-damaging oviposition behavior of this species.

Wild African Drosophila melanogaster Are Seasonal Specialists on Marula Fruit.

Although the vinegar fly Drosophila melanogaster is arguably the most studied organism on the planet, fundamental aspects of this species' natural ecology have remained enigmatic [1]. We have here investigated a wild population of D. melanogaster from a mopane forest in Zimbabwe. We find that these flies are closely associated with marula fruit (Sclerocarya birrea) and propose that this seasonally abundant and predominantly Southern African fruit is a key ancestral host of D. melanogaster. Moreover, when fruiting, marula is nearly exclusively used by D. melanogaster, suggesting that these forest-dwelling D. melanogaster are seasonal specialists, in a similar manner to, e.g., Drosophila erecta on screw pine cones [2]. We further demonstrate that the main chemicals released by marula activate odorant receptors that mediate species-specific host choice (Or22a) [3, 4] and oviposition site selection (Or19a) [5]. The Or22a-expressing neurons-ab3A-respond strongly to the marula ester ethyl isovalerate, a volatile rarely encountered in high amounts in other fruit. We also show that Or22a differs among African populations sampled from a wide range of habitats, in line with a function associated with host fruit usage. Flies from Southern Africa, most of which carry a distinct allele at the Or22a/Or22b locus, have ab3A neurons that are more sensitive to ethyl isovalerate than, e.g., European flies. Finally, we discuss the possibility that marula, which is also a culturally and nutritionally important resource to humans, may have helped the transition to commensalism in D. melanogaster.

The Scent of the Fly.

(Z)-4-undecenal (Z4-11Al) is the volatile pheromone produced by females of the vinegar fly Drosophila melanogaster. Female flies emit Z4-11Al for species-specific communication and mate-finding. A sensory panel finds that synthetic Z4-11Al has a characteristic flavour, which can be perceived even at the small amounts produced by a single female fly. Since only females produce Z4-11Al, and not males, we can reliably distinguish between single D. melanogaster males and females, according to their scent. Females release Z4-11Al at 2.4 ng/h and we readily sense 1 ng synthetic Z4-11Al in a glass of wine (0.03 nmol/L), while a tenfold concentration is perceived as a loud off-flavour. This corroborates the observation that a glass of wine is spoilt by a single D. melanogaster fly falling into it, which we here show is caused by Z4-11Al. The biological role of Z4-11Al or structurally related aldehydes in humans and the basis for this semiochemical convergence remains yet unclear.

Chemical signaling and insect attraction is a conserved trait in yeasts.

Yeast volatiles attract insects, which apparently is of mutual benefit, for both yeasts and insects. However, it is unknown whether biosynthesis of metabolites that attract insects is a basic and general trait, or if it is specific for yeasts that live in close association with insects. Our goal was to study chemical insect attractants produced by yeasts that span more than 250 million years of evolutionary history and vastly differ in their metabolism and lifestyle. We bioassayed attraction of the vinegar fly Drosophila melanogaster to odors of phylogenetically and ecologically distinct yeasts grown under controlled conditions. Baker's yeast Saccharomyces cerevisiae, the insect-associated species Candida californica, Pichia kluyveri and Metschnikowia andauensis, wine yeast Dekkera bruxellensis, milk yeast Kluyveromyces lactis, the vertebrate pathogens Candida albicans and Candida glabrata, and oleophilic Yarrowia lipolytica were screened for fly attraction in a wind tunnel. Yeast headspace was chemically analyzed, and co-occurrence of insect attractants in yeasts and flowering plants was investigated through a database search. In yeasts with known genomes, we investigated the occurrence of genes involved in the synthesis of key aroma compounds. Flies were attracted to all nine yeasts studied. The behavioral response to baker's yeast was independent of its growth stage. In addition to Drosophila, we tested the basal hexapod Folsomia candida (Collembola) in a Y-tube assay to the most ancient yeast, Y. lipolytica, which proved that early yeast signals also function on clades older than neopteran insects. Behavioral and chemical data and a search for selected genes of volatile metabolites underline that biosynthesis of chemical signals is found throughout the yeast clade and has been conserved during the evolution of yeast lifestyles. Literature and database reviews corroborate that yeast signals mediate mutualistic interactions between insects and yeasts. Moreover, volatiles emitted by yeasts are commonly found also in flowers and attract many insect species. The collective evidence suggests that the release of volatile signals by yeasts is a widespread and phylogenetically ancient trait, and that insect-yeast communication evolved prior to the emergence of flowering plants. Co-occurrence of the same attractant signals in yeast and flowers suggests that yeast-insect communication may have contributed to the evolution of insect-mediated pollination in flowers.