Pollinator Preferences for Floral Volatiles Emitted by Dimorphic Anthers of a Buzz-Pollinated Herb.

Floral scents attract pollinators to plant rewards; in nectarless flowers, pollen grains are the only reward. Thus, pollen not only fertilizes ovules, but also feeds pollinators. This dilemma is resolved by specialization of anthers (i.e., heteranthery): feeding anthers that feed pollinators and pollinating anthers for fertilization. We hypothesized that the chemical composition of floral volatiles differs between the two types of anther and influences pollination preference for feeding anthers. We used Solanum rostratum as a study model because its heterantherous flowers produce a floral scent that suggests a close association with their pollinators. The main aim of this study was to determine the chemical composition of the two types of anther and to investigate how they influence foraging behaviour of pollinators. To characterize this composition, we used solid phase microextraction and hexane extraction followed by gas chromatography-mass spectrometry. We registered 12 volatile compounds in S. rostratum floral extracts, mainly aromatic and sesquiterpene compounds. The proportion of these compounds differed between feeding and pollinating anthers. Some of these compounds were probably emitted by osmophores located in both anther types. Also, we used electroantennography to investigate Melipona solani antennal response to floral volatiles. The M. solani antennae are receptive to the highest floral extract dose tested. Finally, we conducted two behavioural bioassays to test bee attraction for each type of floral extract: a) multiple-choice in a feeding arena using M. solani and b) Y-olfactometer bioassay using Bombus impatiens. Both bee species preferred feeding anthers in bioassays. In conclusion, heteranthery involves chemical differentiation (i.e., proportion of volatiles compounds) in anther specialization that influences bee preference for feeding anthers over pollinating anthers.

Behavioral and Antennal Responses of Drosophila suzukii (Diptera: Drosophilidae) to Volatiles From Fruit Extracts.

Native to Southeast Asia, the spotted wing drosophila, Drosophila suzukii Matsumura (Diptera: Drosophilidae), has become a serious pest of soft-skinned fruit crops since its introduction into North America and Europe in 2008. Current monitoring strategies use baits based on fermentation products; however, to date, no fruit-based volatile blends attractive to this fly have been identified. This is particularly important because females are able to cut into the epicarp of ripening fruit for oviposition. Thus, we conducted studies to: 1) investigate the behavioral responses of adult D. suzukii to volatiles from blueberry, cherry, raspberry, and strawberry fruit extracts; 2) identify the antennally active compounds from the most attractive among the tested extracts (raspberry) using gas chromatography (GC)-mass spectrometry and coupled gas chromatography -electroantennographic detection (GC-EAD); and 3) test a synthetic blend containing the EAD-active compounds identified from raspberry extract on adult attraction. In olfactometer studies, both female and male D. suzukii were attracted to all four fruit extracts. The attractiveness of the fruit extracts ranks as: raspberry ≥ strawberry > blueberry ≥ cherry. GC analyses showed that the fruit extracts emit distinct volatile compounds. In GC-EAD experiments, 11 raspberry extract volatiles consistently elicited antennal responses in D. suzukii. In choice test bioassays, a synthetic EAD-active blend attracted more D. suzukii than a blank control, but was not as attractive as the raspberry extract. To our knowledge, this is the first report of a behaviorally and antennally active blend of host fruit volatiles attractive to D. suzukii, offering promising opportunities for the development of improved monitoring and behaviourally based management tools.

Behavioral and electroantennogram responses of plum curculio, Conotrachelus nenuphar, to selected noxious plant extracts and insecticides.

Behavioral and electroantennogram responses of plum curculio, Conotrachelus nenuphar (Herbst) (Coleoptera: Curculionidae), adults were tested for several methanolic plant extracts and organically approved insecticides. Plant extracts were evaluated for their potential as antifeedants or oviposition deterrents. These extract responses were also compared to those elicited by the non-neurotoxic, organic irritant-insecticide kaolin clay. Both sexes of plum curculio exhibited antennal response as measured by electroantennogram, which ranged from 0.2 to 1.1 mV, to plant extracts and the organic irritant/insecticide, with the greatest response to the extract of rough cocklebur, Xanthium strumarium L. (1.1 mV). No choice tests were conducted to compare feeding and oviposition by plum curculio on untreated apples or on apples treated with one of the extracts or the insecticide. The insecticide pyrethrum and extracts of X. strumarium and greater burdock, Arctium lappa L., significantly reduced feeding. Also, pyrethrum, A. lappa, Humulus lupulus L. (common hop), X. strumarium, and Verbascum songaricum Schrenk extracts completely inhibited egg deposition. In no-choice assays, the effects of kaolin clay with incorporated plant extracts on plum curculio feeding and oviposition were monitored as complementary tests. A. lappa-kaolin, H. lupulus-kaolin, and X. strumarium-kaolin mixtures significantly reduced the feeding of plum curculio compared to the control or kaolin clay alone. Each of the plant extract-kaolin mixtures evaluated, with the exception of Bifora radians Bieberstein (wild bishop), completely inhibited plum curculio oviposition as compared to controls.

EAG responses increase of Spodoptera littoralis antennae after a single pheromone pulse.

Increased behavioral sensitivity to the pheromone after brief exposure of the whole insect to the sex pheromone has been documented in antennal lobe neurons of Spodoptera littoralis. We investigated whether a brief stimulus of the major component of the pheromone on naïve antenna separated from the head increased the electroantennographic responses after successive stimulations at different times. The response increase was clear 30 min after the first stimulation, and this effect lasted at least 60 min, the average life time of the antenna. Our results suggest that the olfactory receptor neurons, and not only the neurons in the antennal lobe, may be involved in the increased antennal response after a single pheromone pulse.

Chemical composition, larvicidal action, and adult repellency of Thymus magnus against Aedes albopictus.

Thymus magnus, an endemic species, is found in the Republic of Korea. The volatile compounds extracted by SPME from T. magnus were investigated for their chemical composition and electrophysiological response against the Asian tiger mosquito, Aedes albopictus. The volatile compounds of T. magnus as determined by gas chromatography mass spectrometry were gamma-terpinene (33.0%), thymol (29.9%), beta-bisabolene (8.9%), p-cymene (8.3%), alpha-terpinene (5.0%), myrcene (4.7%), beta-caryophyllene (4.0%), alpha-thujene (2.7%), camphene (1.3%), carvacrol (1.2%), and alpha-pinene (1.1%). Among these candidates, thymol exhibited complete (100%) repellent activity against female Ae. albopictus, an effect that was confirmed through evaluating the electrophysiological response on the antenna of Ae. albopictus. The effectiveness of a binary 1:2 mixture of thymol and vanillin (0.05:0.1 microl per cm2) was found to be significantly more effective than thymol alone for a period of 120 min. In addition, thymol, alpha-terpinene, and carvacrol showed high larvicidal activity against on the third-stage larvae with LC50 values of 0.9 microl per 100 ml.

Host plant volatiles induce oriented flight behaviour in male European grapevine moths, Lobesia botrana.

The European grapevine moth Lobesia botrana relies on a female produced sex pheromone for long-distance mate finding. Grapevine moth males compete heavily during limited time windows for females. The aim of this study was to investigate the perception of host plant volatiles by grapevine moth males and whether such compounds elicit upwind oriented flights. We compared five host plant headspace extracts by means of gas chromatography linked electroantennogram (EAG) recording. We identified 12 common host plant volatiles (aliphatic esters, aldehydes, and alcohols, aromatic compounds and terpenes) that elicit EAG responses from grapevine moth males and that occur in at least three of the host plant volatile headspace extracts tested. Subsequently the behavioural response of grapevine moth males to four these compounds presented singly and in mixtures (1-hexanol, 1-octen-3-ol, (Z)-3-hexenyl acetate and (E)-ß-caryophyllene) was recorded in a wind tunnel. Grapevine moth males engaged in upwind flights to all of four compounds when released singly at 10,000 pg/min and to all, except 1-octen-3-ol, when released at 100 pg/min. A blend of the four host plant volatiles released at 10,000 pg/min and mixed at a ratio based on the analysis of Vitis vinifera cv. Solaris volatile emissions attracted significantly more males than any single compound. Grapevine moth males perceive and respond to host plant volatiles at biologically relevant levels indicating that host plant volatiles figure as olfactory cues and that L. botrana males can discern places where the likelihood of encountering females is higher.

Switching attraction to inhibition: mating-induced reversed role of sex pheromone in an insect.

In the moth, Agrotis ipsilon, newly mated males cease to be attracted to the female-produced sex pheromone, preventing them from re-mating until the next night, by which time they would have refilled their reproductive glands for a potential new ejaculate. The behavioural plasticity is accompanied by a decrease in neuron sensitivity within the primary olfactory centre, the antennal lobe (AL). However, it was not clear whether the lack of the sexually guided behaviour results from the absence of sex pheromone detection in the ALs, or if they ignore it in spite of detection, or if the sex pheromone itself inhibits attraction behaviour after mating. To test these hypotheses, we performed behavioural tests and intracellular recordings of AL neurons to non-pheromonal odours (flower volatiles), different doses of sex pheromone and their mixtures in virgin and newly mated males. Our results show that, although the behavioural and AL neuron responses to flower volatiles alone were similar between virgin and mated males, the behavioural response of mated males to flower odours was inhibited by adding pheromone doses above the detection threshold of central neurons. Moreover, we show that the sex pheromone becomes inhibitory by differential central processing: below a specific threshold, it is not detected within the AL; above this threshold, it becomes inhibitory, preventing newly mated males from responding even to plant odours. Mated male moths have thus evolved a strategy based on transient odour-selective central processing, which allows them to avoid the risk-taking, energy-consuming search for females and delay re-mating until the next night for a potential new ejaculate.