Identification and analysis of chemosensory genes encoding odorant-binding proteins, chemosensory proteins and sensory neuron membrane proteins in the antennae of Lissorhoptrus oryzophilus

The rice water weevil, Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae), is a destructive pest that causes damage to rice crops worldwide. The olfactory system is critical for host or mate location by weevils, but only limited information about the molecular mechanism of olfaction-related behaviour has been reported in this insect. In this study, we conducted SMRT-seq transcriptome analysis and obtained 54,378 transcripts, 38,706 of which were annotated. Based on these annotations, we identified 40 candidate chemosensory genes, including 31 odorant-binding proteins (OBPs), six chemosensory proteins (CSPs) and three sensory neuron membrane proteins (SNMPs). Phylogenetic analysis showed that LoryOBPs, LoryCSPs and LorySNMPs were distributed in various clades. The results of tissue expression patterns indicated that LoryOBPs were highly abundant in the antennae, whereas LoryCSPs were highly abundant not only in the antennae but also in the abdomen, head and wings. Our findings substantially expand the gene database of L. oryzophilus and may serve as a basis for identifying novel targets to disrupt key olfactory genes, potentially providing an eco-friendly strategy to control this pest in the future.

Long- and short-range bimodal signals mediate mate location and recognition in yellow fever mosquitoes

As recently reported, light flashes of incident sunlight reflecting off the wings of in-flight dipterans serve as mate recognition signals. Mate location and mate selection behavior in the yellow fever mosquito, Aedes aegypti, take place in mating swarms but the mechanisms underlying swarm formation and long-range detection of females by males remain largely unexplored. Here we show that swarm formation and mate recognition are mediated, in part, by light flash signals and wingbeat sound signals that operate at long and short range, respectively. To test for range-dependent effects of these signals, we presented ‘mating swarms’ in form of two paired 8-LED assemblies that were fitted with micro-speakers and placed either well separated in a large space or side-by-side in a small space. In the large but not the small space, the LED assembly flashing light at the wingbeat frequency of females (665 Hz), and emitting their wingbeat sound (665 Hz), attracted and prompted 5.8-times more alightings by males than the LED assembly emitting constant light and wingbeat sound. In the small space, the LED assembly flashing light and emitting wingbeat sound induced 5.0-times more alightings by males than the LED assembly flashing light without wingbeat sound. Females responded to light flash signals of males, but males failed to respond to the synthetic female pheromone component ketoisophorone added to the bimodal complex of light and sound signals. The attractiveness of light flash signals to males increased with increasing numbers of signals but did not vary according to their wavelengths (UV or blue). As predicted by the sensory drive theory, light flashes had no signal function for crepuscular house mosquitoes, Culex pipiens.

Antennal Transcriptome Analysis and Identification of Candidate Chemosensory Genes of the Harlequin Ladybird Beetle, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae)

In predatory ladybirds (Coleoptera: Coccinellidae), antennae are important for chemosensory reception used during food and mate location, and for finding a suitable oviposition habitat. Based on NextSeq 550 Illumina sequencing, we assembled the antennal transcriptome of mated Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) males and females and described the first chemosensory gene repertoire expressed in this species. We annotated candidate chemosensory sequences encoding 26 odorant receptors (including the coreceptor, Orco), 17 gustatory receptors, 27 ionotropic receptors, 31 odorant-binding proteins, 12 chemosensory proteins, and 4 sensory neuron membrane proteins. Maximum-likelihood phylogenetic analyses allowed to assign candidate H. axyridis chemosensory genes to previously described groups in each of these families. Differential expression analysis between males and females revealed low variability between sexes, possibly reflecting the known absence of relevant sexual dimorphism in the structure of the antennae and in the distribution and abundance of the sensilla. However, we revealed significant differences in expression of three chemosensory genes, namely two male-biased odorant-binding proteins and one male-biased odorant receptor, suggesting their possible involvement in pheromone detection. Our data pave the way for improving the understanding of the molecular basis of chemosensory reception in Coccinellidae.

Mate Choice Behavior of Female Field Crickets Is Not Affected by Exposure to Heterospecific Calling Songs

Many animals produce acoustic signals to mark territories and attract mates. When different species produce acoustic signals simultaneously, the signals create a noisy environment, with potential acoustic interference between species. Theoretical studies suggest that such reproductive interference may have strong effects on species interaction. For example, the inferior resource competitor can survive if its disadvantage is counterbalanced by superiority in reproductive interference. Two field cricket species, Teleogryllus occipitalis (Audinet-Serville) (Orthoptera: Gryllidae) and Loxoblemmus equestris Saussure (Orthoptera: Gryllidae), cooccur in the same habitat. A previous study has shown that L. equestris is an inferior species to T. occipitalis in terms of resource competition. Therefore, we predicted that mate location and choice behavior of female T. occipitalis would be negatively affected by the acoustic signals of L. equestris and tested this with a series of playback experiments. The mate choice behavior of female T. occipitalis was not significantly affected by the calling song of L. equestris. Our results suggest that the acoustic interference does not explain the cooccurrence of the two species in the same habitat.

The Role of (E)-2-octenyl Acetate as a Pheromone of Bagrada hilaris (Burmeister): Laboratory and Field Evaluation

The pentatomid bug Bagrada hilaris is a key pest of brassicaceous crops in several areas of the world. Previous studies suggest that mate location of this species is mediated by volatile chemicals produced by males, among which the main compound is (E)-2-octenyl acetate. However, the possible attraction of males, females, and nymphs to this compound has not yet been specifically tested. In this study, we tested the response of B. hilaris females, males, and nymphs to (E)-2-octenyl acetate using an electroantennogram (EAG) and olfactometer in the presence or absence of a host plant. Moreover, (E)-2-octenyl acetate as an attractant lure in field trap bioassays was evaluated. EAG recordings showed that this compound evokes antennal responses in B. hilaris females. Olfactometer behavioral responses showed that females and nymphs were attracted to (E)-2-octenyl acetate, while males showed no attraction. In the field trap bioassays, captures were obtained in traps baited with 5 and 10 mg of (E)-2-octenyl acetate, while in traps loaded with 2 mg and control traps, there were no recorded catches. These results suggest the involvement of (E)-2-octenyl acetate in intraspecific interactions of this species.

Multiple Mating in the Citrophilous Mealybug Pseudococcus calceolariae: Implications for Mating Disruption

The citrophilous mealybug Pseudococcus calceolariae (Maskell) (Hemiptera, Pseudococcidae) is a primary pest of various crops, including grapevines. The use of insecticides against this species is difficult in most cases because its life cycle includes an extended duration of eggs, juveniles, and adults under the bark and on the roots. Pheromone-based control strategies can present new eco-friendly opportunities to manage this species, as in the case of Planococcus ficus (Signoret) and Planococcus citri (Risso). With this aim it is critical to understand behavioral aspects that may influence pheromone-based control strategies. Herein, the capability of males to fertilize multiple females was investigated, trying to understand whether this behavior could negatively impact the efficacy of mass trapping, mating disruption, or the lure and kill technique. Results showed that a P. calceolariae male can successfully mate and fertilize up to 13 females. The copulation time in subsequent mating events and the time between copulations did not change over time but the number of matings per day significantly decreased. In a further experiment, we investigated the mate location strategy of P. calceolariae males, testing the attractiveness of different loadings of sex pheromone on males in a flight tunnel. Males constantly exposed to 16 rubber septa loaded with the sex pheromone showed a significant decrease in female detection at 1 and 30 μg loadings (0.18 and 0.74 visits per female for each visit per septum, respectively), whereas in the control about 9.2-fold more of the released males successfully detected the female in the center of the array of 16 septa without pheromone. Male location of females in the control (45%) was significantly higher than in the arrays with surrounding pheromone (5% and 20% at 1 and 30 μg loadings, respectively). Mating only occurred in the control arrays (45%). This study represents a useful first step to developing pheromone-based strategies for the control of citrophilous mealybugs.

Different male mate location behaviour of the Glanville fritillary butterfly in different landscapes in the Tianshan Mountains, northwestern China

In a previous study most males of the Glanville fritillary butterfly (Melitaea cinxia) were caught in valleys, whereas almost all females were distributed on slopes in the Tianshan Mountains, northwestern China. To help understand this phenomenon, male mate location behaviours were observed in different landscapes of the Tianshan Mountains. In valleys, males exhibited perching behaviour. On slopes, spatial distribution of males showed patrolling behaviour on meadows, but intermediate behaviour between perching and patrolling at forest edge. The temporal distribution of males also varied, being found on slopes from 7:00 to 18:00, but in valleys from 8:00 to 13:00 each day. Ambient temperatures were higher on slopes than those in valleys between 8:00 to 13:00. Males exhibited lower tolerance to high temperature than females, leading to the conclusion that valleys are more likely to be used by males as thermoregulation sites, rather than for mating.