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Grosse-Wilde, E.

Publications and source records attributed to Grosse-Wilde, E..

9 recordsLinked to original sources

Identification of the trail-following pheromone receptor in termites

Chemical communication is the cornerstone of eusocial insect societies since it mediates the social hierarchy, division of labor, and concerted activities of colony members. The chemistry of social insect pheromones received considerable attention in both major groups of social insects, the eusocial Hymenoptera and termites. By contrast, current knowledge on molecular mechanisms of social insect pheromone detection by odorant receptors (ORs) is limited to hymenopteran social insects and no OR was yet functionally characterized in termites, the oldest eusocial insect clade. Here, we present the first OR deorphanization in termites. We selected four OR sequences from the previously annotated antennal transcriptome of the termite Prorhinotermes simplex (Rhinotermitidae), expressed them in Empty Neuron Drosophila, and functionally characterized using single sensillum recording (SSR) and a panel of termite semiochemicals. In one of the selected ORs, PsimOR14, we succeeded in obtaining strong and reliable responses to the main component of P. simplex trail-following pheromone, the monocyclic diterpene neocembrene. PsimOR14 showed a narrow tuning to neocembrene; only one additional compound out of 67 tested (geranylgeraniol) generated non-negligible responses. Subsequently, we used SSR in P. simplex workers and identified the olfactory sensillum specifically responding to neocembrene, thus likely expressing PsimOR14. We report on homology-based modelling of neocembrene binding by PsimOR14 and show how different ligands impact the receptor dynamicity using molecular dynamics simulations. Finally, we demonstrate that PsimOR14 is significantly more expressed in worker antennae compared to soldiers, which correlates with higher sensitivity of workers to neocembrene.

animal behavior and cognition↗

Sensing volatiles throughout the body: Geographic and tissue-specific olfactory receptor expression in the fig wasp Ceratosolen fusciceps

An essential adaptive strategy in insects is the evolution of olfactory receptors (ORs) to recognize important volatile environmental chemical cues. Our model species, Ceratosolen fusciceps, a specialist wasp pollinator of Ficus racemosa, likely possesses an OR repertoire that allows it to distinguish fig-specific volatiles in highly variable environments. Using a newly assembled genome-guided transcriptome, we annotated 63 ORs in the species and reconstructed the phylogeny of Ceratosolen ORs in conjunction with other hymenopteran species. Expression analysis showed that though ORs were mainly expressed in the antennae, 20 percent were also expressed in non-antennal tissues such as the head, thorax, abdomen, legs, wings, and ovipositor. Specific upregulated expression was observed in OR30C in the head and OR60C in the wings. We identified OR expression from all major body parts of C. fusciceps, suggesting novel roles of ORs throughout the body. Further examination of OR expression of C. fusciceps in widely separated geographical locations, i.e., south (urban) and northeast (rural) India, revealed distinct OR expression levels in different locations. This discrepancy likely parallels the observed variation in fig volatiles between these regions and provides new insights into the evolution of insect ORs and their expression across geographical locations and tissues.

genomics↗

Odorant receptor orthologues in conifer-feeding beetles display conserved responses to ecologically relevant odors

Insects are able to detect a plethora of olfactory cues using a divergent family of odorant receptors (ORs). Despite the divergent nature of this family, related species frequently express several evolutionarily conserved OR orthologues. In the largest order of insects, Coleoptera, it remains unknown whether OR orthologues have conserved or divergent functions in different species. Using HEK293 cells, we addressed this question through functional characterization of two groups of OR orthologues in three species of the Curculionidae (weevil) family, the conifer-feeding bark beetles Ips typographus L. ( Ityp) and Dendroctonus ponderosae Hopkins ( Dpon) (Scolytinae), and the pine weevil Hylobius abietis L. ( Habi; Molytinae). The ORs of H. abietis were annotated from antennal transcriptomes. Results show highly conserved response specificities, with one group of orthologues (HabiOR3/DponOR8/ItypOR6) responding exclusively to 2-phenylethanol (2-PE), and the other group (HabiOR4/DponOR9/ItypOR5) responding to angiosperm green leaf volatiles (GLVs). Both groups of orthologues belong to the coleopteran OR subfamily 2B, and share a common ancestor with OR5 in the cerambycid Megacyllene caryae, also tuned to 2-PE, suggesting a shared evolutionary history of 2-PE receptors across two beetle superfamilies. The detected compounds are ecologically relevant for conifer-feeding curculionids, and are probably linked to fitness, with GLVs being used to avoid angiosperm non-host plants, and 2-PE being important for intraspecific communication and/or playing a putative role in beetle-microbe symbioses. To our knowledge, this study is the first to reveal evolutionary conservation of OR functions across several beetle species and hence sheds new light on the functional evolution of insect ORs.

neuroscience↗

Changes in antennal gene expression underlying sensory system maturation in Rhodnius prolixus

BackgroundTriatomine bugs are the blood feeding insect vectors transmitting Chagas disease to humans, a neglected tropical disease that affects over 8 million people, mainly in Latin America. The behavioral responses to host cues and bug signals in Rhodnius prolixus are state dependent, i.e., they vary as a function of post-ecdysis age. At the molecular level, these changes in behavior are probably due to a modulation of peripheral and central processes. In the present study, we report a significant modulation of the expression of a large set of sensory-related genes. Results were generated by means of antennal transcriptomes of 5th instar larvae along the first week (days 0, 2, 4, 6 and 8) after ecdysis sequenced using the Illumina platform. ResultsAge induced significant changes in transcript abundance were established in more than 6,120 genes (54,7 % of 11,186 genes expressed) in the R. prolixus antenna. This was especially true between the first two days after ecdysis when more than 2,500 genes had their expression significantly altered. In contrast, expression profiles were almost identical between day 6 and 8, with only a few genes showing significant modulation of their expression. A total of 86 sensory receptors, odorant carriers and odorant degrading enzymes were significantly modulated across age points and clustered into three distinct expression profiles. ConclusionsThe set of sensory genes whose expression increased with age (profile 3) may include candidates underlying the increased responsiveness to host cues shown by R. prolixus during the first days after molting. For the first time, we describe the maturation process undergone at the molecular level by the peripheral sensory system is described in an hemimetabolous insect.

molecular biology↗

A highly contiguous genome assembly of a major forest pest, the Eurasian spruce bark beetle Ips typographus

The Eurasian spruce bark beetle (Ips typographus [L.]), is a major killer of spruce forests across the Palearctic. During epidemics, it can destroy over 100 million cubic meters of spruce trees in a single year. Here we report a 236 Mb, highly contiguous I. typographus genome assembly using PacBio long-read sequencing. The final phased assembly had a contig N50 of 6.65 Mb in 272 contigs and was predicted to contain 23,923 protein-coding genes. Comparative genomic analysis revealed expanded gene families associated with plant cell wall degradation, including pectinases, aspartyl proteases, and glycosyl hydrolases. In todays forests, increasingly stressed by global warming, this resource can assist in mitigating bark beetle outbreaks by developing novel pest control strategies. Further, this first whole-genome sequence from the genus Ips provides timely resources to address important questions about the evolutionary biology and ecology of Curculionidae, the true weevils, one of the largest animal families.

genomics↗

Putative ligand binding sites of two functionally characterized bark beetle odorant receptors

AO_SCPLOWBSTRACTC_SCPLOWBark beetle behavior is to a large extent mediated via olfaction. Targeting the odorant receptors (ORs) may thus provide avenues towards improved pest control during outbreaks. Such an approach requires information on the function of receptors and their interactions with ligands. Hence, we annotated 73 ORs from an antennal transcriptome of the spruce bark beetle Ips typographus and report the functional characterization of two ORs (ItypOR46 and ItypOR49), which are selective for single enantiomers of the common bark beetle pheromone compounds ipsenol and ipsdienol, respectively. We use homology modeling and molecular docking to predict their binding sites. The importance of residues Tyr84 and Thr205 in ItypOR46 in the activation by ipsenol is experimentally supported, and hydrogen bonding appears key in pheromone binding. The biological significance of the characterized ORs positions them as prime targets for pest control and use in biosensors to detect bark beetle infestations.

neuroscience↗

The olfactory co-receptor IR8a governs larval-frass mediated competition avoidance in a hawkmoth

Finding a suitable oviposition site is a challenging task for a gravid female moth. At the same time, it is of paramount importance considering the limited capability of most caterpillars to relocate to alternative host plants. The hawkmoth, Manduca sexta (Sphingidae), oviposits on solanaceous plants. Larvae hatching on a plant that is already attacked by conspecific caterpillars can face food competition, as well as an increased exposure to predators and induced plant defenses. Here, we show that frass from conspecific caterpillars is sufficient to deter a female M. sexta from ovipositing on a plant and that this deterrence is based on the frass-emitted carboxylic acids 3-methylpentanoic acid and hexanoic acid. Using a combination of genome editing (CRISPR/Cas9), electrophysiological recordings, calcium imaging and behavioral analyses we demonstrate that the ionotropic co-receptor IR8a is essential for acid-mediated frass avoidance in ovipositing hawkmoths.

neuroscience↗

Transcriptome Surveys in Silverfish Suggest a Multistep Origin of the Insect Odorant Receptor Gene Family

The insect odorant receptor (Or) gene family is among the largest multigene families in insect genomes, but its evolutionary origin and mode of expansion is still a matter of debate. We performed transcriptomic surveys of two wingless insect species, the silverfish Lepisma saccharina and Tricholepidion gertschi, and identified multiple Or gene family members in both species. A phylogenetic analysis suggests that the silverfish Ors do not fall into the clade comprised of more derived flying insect ligand-binding Ors, but, along with bristletail, firebrat and some mayfly Ors, are consistently resolved as a distinct set of genes that may constitute an evolutionary intermediate between gustatory receptors and the more derived Ors of flying insects. We propose to consider these \"primitive Ors\" separately from higher insect Ors until their cellular expression patterns and function are resolved and suggest a multistep evolutionary scenario ultimately leading to the highly sensitive, rapidly evolving and physiologically diverse Or gene family observed in higher insects.

evolutionary biology↗

Mutagenesis of orco impairs foraging but not oviposition in the hawkmoth Manduca sexta

Plant volatile detection through olfaction plays a crucial role in insect behaviors. In vivo, the odorant receptor co-receptor orco is an obligatory component for the function of odorant receptors (ORs), a major receptor family involved in insect olfaction. We used CRISPR-Cas9 targeted mutagenesis to knock-out (KO) orco in a neurophysiological model species, the hawkmoth Manduca sexta. M. sexta and its host, the Sacred Datura (Datura wrightii) share a model insect-plant relationship based on mutualistic and antagonistic life history traits. D. wrightii is the innately preferred nectar-source and oviposition host for M. sexta. Hence, the hawkmoth is an important pollinator while the M. sexta larvae are specialized herbivores of the plant. We generated an orco KO through CRISPR-Cas9 to test the consequences of a loss of OR-mediated olfaction in this insect-plant relationship. Neurophysiological characterization revealed severely reduced antennal and antennal lobe responses to representative odorants emitted by D. wrightii. In a wind-tunnel setting with a flowering plant, orco KO hawkmoths showed disrupted flight orientation and an ablated proboscis extension response to the natural stimulus. However, when testing the oviposition behavior of mated females encountering a non-flowering plant, there was no difference between orco KO and wild type females regarding upwind flight orientation and number of eggs laid. Overall, OR-mediated olfaction is essential for foraging and pollination behaviors, but plant-seeking and oviposition behaviors appear largely unaffected. Significance statementInsects detect plant volatiles mainly through the expression of ORs and IRs on the antennal olfactory sensory neurons (OSNs). In vivo, Orco is an obligate partner for OR, but not IR function and ORs mediate a vast spectrum of olfactory perception. We applied CRISPR-Cas9 in M. sexta to mutate the orco gene and determine the physiological and behavioral implication of a loss of Orco receptor function in a semi-ecological interaction with D. wrightii. We found that while behaviors related to foraging were largely disrupted, other sensory modalities outside Orco function determine the relationship between an ovipositing female and its plant host. These results have implications toward understanding the olfactory basis of insect-plant interactions shaping our ecological and agricultural landscapes.

neuroscience↗