A comparison of color-, shape- and pattern-learning by the hymenopteran parasitoid Microplitis croceipes

Visual learning by the larval parasitoid, Microplitis croceipes Cresson (Hymenoptera: Braconidae) was investigated in flight chamber studies. During conditioning, free-ranging parasitoids were given a choice between two visual alternatives, only one of which offered a host larva. By using alternatives that differed in either color, shape or pattern, parasitoids could be conditioned to distinguish host sites on the basis of each of these visual cues. Tests during which no reward was offered were conducted following six rewards (ovipositions) at one of the two alternative stimuli. The test results reveal that M. croceipes learned to distinguish between shapes more readily than between colors or patterns. This high rate of shape learning in this parasitoid is in strong contrast to the learning capacity of honey bees, which have been shown to learn color better than pattern and pattern better than shapes. It is argued that the difference in learning capacities between M. croceipes and the honey bee may reflect the different selection pressures imposed on these two species by their natural ecological needs. The high rate of shape learning in  M. croceipes may be adaptive in dealing with the homochromatic but multishaped environment in which parasitoids have to locate their herbivorous hosts. Accepted: 30 January 1999     

Synergism of pheromone and host‐plant volatile blends in the attraction of Grapholita molesta males

Control of Grapholita molesta (Busck) (Lepidoptera: Tortricidae), a major pest of stone and pome fruits, is successfully achieved by mating disruption. Under these conditions, tools other than conventional pheromone dispensers are needed for flight monitoring. The objective of the present work was to determine whether plant volatiles synergize male G. molesta attraction to a suboptimal dose of synthetic sex pheromone. The plant blend (referred to as 5VB), a mixture of three green leaf volatiles [(Z)‐3‐hexenyl acetate, (Z)‐3‐hexenol, and (E)‐2‐hexenal] and two aromatics [benzaldehyde (BZA) and benzonitrile (BZN)], was added to the suboptimal pheromone dose (2 ng on filter paper) in log steps (up to 10 000× the pheromone dose) to test synergism of pheromone and plant blends. In addition, the effect of individual plant volatiles on male responses was investigated by adding to the suboptimal pheromone dose each of the four‐compound plant‐volatile blends, resulting from eliminating one volatile from the 5VB at a time, or each plant volatile alone. Flight behaviour and the time to reach the source were recorded. The 5VB alone was not attractive to G. molesta males, but at a ratio of 1:1 000 (Ph:5VB) or higher, the attractiveness of the suboptimal pheromone dose increased, to a level similar to that of the optimal pheromone dose (10 ng). All tested plant volatiles, except BZA, synergized the response to the pheromone when added individually, but only (Z)‐3‐hexenol and BZN did so to a level not significantly different from the Ph:5VB blend. Aromatics had a stronger effect than green leaf volatiles (GLVs), because their removal, but not the removal of GLVs, decreased landing responses. The addition of the 5VB decreased significantly the time males needed to reach the odour source. The observed enhanced male attraction to mixtures of pheromone and plant volatiles will facilitate the development of lures for G. molesta adult flight monitoring.     

Variability in response specificity of apple, hawthorn, and flowering dogwood-infesting Rhagoletis flies to host fruit volatile blends: implications for sympatric host shifts

Rhagoletis pomonella Walsh (Diptera: Tephritidae) originating from domesticated apple (Malus pumila), hawthorn (Crataegus mollis) (Rosaceae), and flowering dogwood (Cornus florida) (Cornaceae) were tested sequentially in flight‐tunnel assays to volatile blends previously identified from the three fruit types. The majority of flies flew to odor sources containing their natal blend (68–83%). Some flies from each fruit type also flew to non‐natal fruit blends (11–39%), but of these non‐natal responders the vast majority were flies that responded to their natal blend as well. The results indicate that individual flies within R. pomonella populations infesting different host types have different degrees of specificity with respect to discriminating among fruit volatile blends, and that a moderate proportion of apple, hawthorn, and dogwood flies (10–30%) are broad responders, with the capacity to recognize and orient to more than one blend. The observed variability in response specificity could facilitate sympatric shifts to new host plants.     

Response to host plant odors and aggregation pheromone by larvae of the Colorado potato beetle on a servosphere

Using a servosphere (locomotion compensator), locomotory behavior of Colorado potato beetle Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae) larvae was measured in detail in response to pulsed and non-pulsed odors of hostplant and conspecific pheromone. Second instars showed decreased straightness of movement, and all larvae showed decreased speed, in response to potato odor. Change in straightness by 2nd instars was also significantly affected by the interaction of pheromone and pulsing treatments. Fourth instars showed increased positive anemotaxis in response to the combined hostplant and pheromone odors. To our knowledge, this is the first demonstration of larval behavioral responses to adult pheromone in a holometabolous insect.     

Body temperature of the parasitic wasp Pimpla turionellae (Hymenoptera) during host location by vibrational sounding

Abstract.  The pupal parasitoid Pimpla turionellae (L.) uses self-produced vibrations transmitted on the plant substrate, so-called vibrational sounding, to locate immobile concealed pupal hosts. The wasps are able to use vibrational sounding reliably over a broad range of ambient temperatures and even show an increased signal frequency and intensity at low temperatures. The present study investigates how control of body temperature in the wasps by endothermic mechanisms may facilitate host location under changing thermal environments. Insect body temperature is measured with real-time IR thermography on plant-stem models at temperature treatments of 10, 18, 26 and 30 °C, whereas behaviour is recorded with respect to vibrational host location. The results reveal a low-level endothermy that likely interferes with vibrational sound production because it occurs only in nonsearching females. At the lowest temperature of 10 °C, the thoracic temperature is 1.15 °C warmer than the ambient surface temperature whereas, at the high temperatures of 26 and 30 ° C, the wasps cool down their thorax by 0.29 and 0.47 °C, respectively, and their head by 0.45 and 0.61 °C below ambient surface temperature. By contrast, regardless of ambient temperature, searching females always have a slightly elevated body temperature of at most 0.30 °C above the ambient surface temperature. Behavioural observations indicate that searching females interrupt host location more frequently at suboptimal temperatures, presumably due to the requirements of thermoregulation. It is assumed that both mechanisms, producing vibrations for host location and low-level endothermy, are located in the thorax. Endothermy by thoracic muscle work probably disturbs signal structure of vibrational sounding, so the processes cannot be used at the same time.     

Parasitoid exploitation of the seasonal variation in host plant volatile emission for herbivore location

Volatile compounds from the apple, Malus domestica Borkh. (Rosaceae), change considerably as the season progresses, and this is successfully exploited by the female codling moth Cydia pomonella L. (Lepidoptera, Tortricidae), as it searches for oviposition sites. In this study, we investigated the effect of seasonal emissions of apple fruit volatiles on the host location behaviour of a parasitoid of the codling moth larvae, Hyssopus pallidus (Askew) (Hymenoptera: Eulophidae). In dual choice olfactory bioassays, the behaviour of the parasitoid in response to apple cues was observed over the complete 2003 growing season. Our results show that codling moth infested apples evoked a strong response from the parasitoid at the beginning of the season, until July. Then, attraction dropped drastically, increasing again at the beginning of August. At the end of the growing season, just prior to harvest, infested apples hardly evoked any behavioural response. Interestingly enough, mid‐season emissions of healthy apples were per se attractive to the parasitoid, and even preferred over volatiles from infested apples. Simultaneous volatile collections from healthy apples on twigs in the field were analysed throughout the season, showing that the overall quantity of headspace volatiles peaks at the beginning of June and mid‐August. The seasonal volatile emission is correlated with the behaviour of the parasitoid during the fruit ripening stage. The results are discussed in relation to the use of H. pallidus as a potential biocontrol agent, in order to enhance current integrated pest management (IPM) programs.     

Identification of volatile compounds released by roots of an invasive plant, bitou bush (Chrysanthemoides monilifera spp. rotundata), and their inhibition of native seedling growth

Allelopathy has been suggested as a mechanism promoting the monoculture formation of some invasive exotic plants. Previous studies have shown that hydrophobic extracts of the roots and soil of exotic bitou bush (Chrysanthemoides monilifera spp. rotundata (DC.) T. Norl.) inhibited the seedling growth of five Australian native plants, including the dominant acacia (Acacia longifolia var. sophorae (Labill.) F. Muell.). Based on this finding, we compared the hydrophobic root and soil chemical profiles of bitou bush and acacia to determine whether bitou bush roots release allelopathic compounds that are novel to the invaded system. We detected three compounds that were exclusive to the bitou bush root and soil, and seven compounds that were common to the bitou bush and acacia roots but only present in the bitou bush soil. The compounds unique to the bitou bush invaded soil were all sesqui- and diterpenes. Several of these compounds were found to inhibit the seedling growth of a native sedge, Isolepis nodosa (Rott.) R. Br. Of particular interest are the sesquiterpenes: β-maaliene, α-isocomene, β-isocomene, δ-cadinene, 5-hydroxycalamenene and 5-methoxycalamenene which were found in high concentrations in the bitou bush root and soil extracts and exhibited phytotoxic activity. Therefore, we present evidence to suggest that bitou bush exudes low molecular weight volatile compounds into the soil which inhibit native plant seedling growth. The reduced establishment of native plants via allelopathy is likely to create space and contribute to the invasion of bitou bush on the eastern Australian coast.     

Between plant and diurnal variation in quantities and ratios of volatile compounds emitted by Vicia faba plants

Ratios of volatile phytochemicals potentially offer a means for insects to recognise their host-plant species. However, for this to occur ratios of volatiles would need to be sufficiently consistent between plants and over time to constitute a host-characteristic cue. In this context we collected headspace samples from Vicia faba plants to determine how consistent ratios of key volatile phytochemicals used in host location by one of its insect pests, the black bean aphid, Aphis fabae, were. These were (E)-2-hexenal, (Z)-3-hexen-1-ol, 1-hexanol, benzaldehyde, 6-methyl-5-hepten-2-one, octanal, (Z)-3-hexen-1-yl acetate, (R)-linalool, methyl salicylate, decanal, undecanal, (E)-caryophyllene, (E)-β-farnesene, (S)-germacrene D, and (E, E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene, which had previously been found to be electrophysiologically and behaviourally active to A. fabae. Although the quantities of volatiles produced by V. faba showed large between plant and diurnal variation, correlations between quantities of compounds indicated that the ratios of certain pairs of volatiles were very consistent. This suggests that there is a host-characteristic cue available to A. fabae in the form of ratios of volatiles.     

Deer browsing reduces leaf damage by herbivorous insects through an induced response of the host plant

We aimed to demonstrate an indirect relationship between a mammalian herbivore (sika deer) and herbivorous insects on the induced responses of a shared host plant, Viburnum dilatatum. Field studies were conducted at three sites (i.e. two islands and one mainland) and within a deer exclusion area. One island, Kinkazan (Kz) Island, harbored a high density of deer while the other sites (controls) had no deer or very low densities of deer. The deer exclusion area had been established approximately 10years earlier on Kz. We collected leaves above the browsing line of the deer and measured leaf hardness and tannin concentration. Leaf damage by insects was used as a measure of insect abundance. Leaves collected at Kz were harder than those from one of the control sites and from inside the deer exclusion area, while no difference was detected among the other controls and inside the exclusion area. In contrast, the tannin concentration of leaves from Kz was lower than in leaves from the control site. Leaf damage by herbivorous insects was lower in Kz than the other study sites. In addition, hole-type leaf damage tended to be higher inside, rather than outside, the exclusion area. These results suggest the possibility that deer browsing increased leaf hardness, which exerted an indirect negative effect on the herbivorous insects utilizing the common host plant. To our knowledge, this is the first study to provide evidence of indirect negative effects between mammalian herbivores and herbivorous insects sharing a host plant.     

Representation of a mixture of pheromone and host plant odor by antennal lobe projection neurons of the silkmoth Bombyx mori

Pheromone-source orientation behavior can be modified by coexisting plant volatiles. Some host plant volatiles enhance the pheromonal responses of olfactory receptor neurons and increase the sensitivity of orientation behavior in the Lepidoptera species. Although many electrophysiological studies have focused on the pheromonal response of olfactory interneurons, the response to the mixture of pheromone and plant odor is not yet known. Using the silkmoth, Bombyx mori, we investigated the physiology of interneurons in the antennal lobe (AL), the primary olfactory center in the insect brain, in response to a mixture of the primary pheromone component bombykol and cis-3-hexen-1-ol, a mulberry leaf volatile. Application of the mixture enhanced the pheromonal responses of projection neurons innervating the macroglomerular complex in the AL. In contrast, the mixture of pheromone and cis-3-hexen-1-ol had little influence on the responses of projection neurons innervating the ordinary glomeruli whereas other plant odors dynamically modified the response. Together this suggests moths can process plant odor information under conditions of simultaneous exposure to sex pheromone.     

The role of host tree condition in attack of white oaks by the twolined chestnut borer,Agrilus bilineatus (Weber) (Coleoptera: Buprestidae)

Summary The twolined chestnut borer, Agrilus bilineatus (Weber) (Coleoptera: Buprestidae), attacks stressed oaks (Quercus spp.) and is associated with extensive mortality of trees in the eastern deciduous forests of North America. We examined host location by the insect and subsequent host mortality in experimentally stressed trees. A. bilineatus adults were able to rapidly and specifically locate stressed oak trees. Up to 160 beetles per week were captured on sticky band traps on the trunks of stressed trees, while beetles rarely landed on unstressed control trees. This suggests that adult borers have an acute perception of host tree quality, and that this perception is from a distance. One mechanism of host location may be detection of volatile compounds produced by stressed trees.The condition of the host tree appears to regulate both beetle attraction and successful colonization. Mortally wounded (xylem-girdled) trees attracted beetles only until the cambium died. Xylem-girdled trees were attacked early in the beetle flight season, but larvae did not survive to emerge as adults from these trees. In contrast, phloemgirdled trees continued to attract beetles throughout the flight period. Phloem-girdled trees which were heavily attacked by A. bilineatus died late in the season in which they were attacked. Lightly attacked trees survived until the following growing season, and were then heavily attacked and killed. In one stand, phloem-girdled trees were not attacked, healed over the girdling wounds and were still alive three years after girdling. These results indicate that oak trees are only attractive to A. bilineatus within a narrow range of physiological conditions following stress but prior to mortality. A. bilineatus appears to be a proximate agent of mortality in stressed oaks in eastern North America.This paper (85-7-8-216) resulted from a project of the Kentucky Agricultural Experiment Station and is published with the approval of the Director     

In situ modification of herbivore-induced plant odors: a novel approach to study the attractiveness of volatile organic compounds to parasitic wasps

Many parasitic wasps (parasitoids) exploit volatile organic compounds (VOCs) emitted by herbivore-infested plants in order to locate their hosts, but it remains largely unknown which specific compounds within the volatile blends elicit the attractiveness to parasitoids. One way of studying the importance of specific VOCs is to test the attractiveness of odor blends from which certain compounds have been emitted. We used this approach by testing the attraction of naive and experienced females of the two parasitoids Cotesia marginiventris and Microplitis rufiventris to partially altered volatile blends of maize seedlings (Zea mays var. Delprim) infested with Spodoptera littoralis larvae. Adsorbing filter tubes containing carbotrap-C or silica were installed in a four-arm olfactometer between the odor source vessels and the arms of the olfactometer. The blends breaking through were tested for chemical composition and attractiveness to the wasps. Carbotrap-C adsorbed most of the sesquiterpenes, but the breakthrough blend remained attractive to naive C. marginiventris females. Silica adsorbed only some of the more polar VOCs, but this essentially eliminated all attractiveness to naive C. marginiventris, implying that among the adsorbed compounds there are some that play key roles in the attraction. Unlike C. marginiventris, M. rufiventris was still attracted to the latter blend, showing that parasitoids with a comparable biology may employ different strategies in their use of plant-provided cues to locate hosts. Results from similar experiments with modified odor blends of caterpillar-infested cowpea (Vigna unguiculata) indicate that key VOCs in different plant species vary greatly in quality and/or quantity. Finally, experienced wasps were more strongly attracted to a specific blend after they perceived the blend while ovipositing in a host. Considering the high number of distinct adsorbing materials available today, this in situ modification of complex volatile blends provides a new and promising approach pinpointing on key attractants within these blends. Advantages and disadvantages compared to other approaches are discussed.     

A tritrophic effect of host plant on susceptibility of western flower thrips to the entomopathogenic fungus Beauveria bassiana

Adult female western flower thrips (Frankliniella occidentalis) were exposed 12-24h to bean (Phaseolus vulgaris) and impatiens (Impatiens wallerana) leaf disks treated with Beauveria bassiana conidia and then transferred to clean bean or impatiens at various times post-treatment. Significantly greater levels of fungal infection were observed when thrips were treated on bean versus impatiens, but exposure to impatiens following treatment had no effect on fungal infection (percent mortality). This result, combined with observations of no inhibition of germination of conidia exposed to intact or macerated impatiens foliage, indicated that the negative effect of the impatiens host plant was not due to plant chemical compounds (antibiosis). Further observations revealed that insects acquired (picked-up) 75% more conidia from treated bean disks than from treated impatiens disks. This difference in dose acquisition was determined to account for the observed difference in percent mortality (15%) following treatment on the two host plants. Median lethal doses (LD(50)) of B. bassiana were not significantly different on the two host plants, but median lethal concentrations were nearly 7-fold greater on impatiens. This difference was explained by disproportionate rates of conidial acquisition at measured rates of conidial deposition (an inverse relationship was observed between application rate expressed as conidia/mm(2) and the number of conidia acquired). The mechanism underlying the differential rates of conidial acquisition from bean versus impatiens was not determined.     

Alterations of the Plasma Membrane Caused by Murine Polyomavirus Proliferation: An Electrorotation Study

In this report we investigate the alterations of the dielectric properties of the plasma membrane caused by the infection of cultured fibroblasts with murine polyomavirus. The approach consists in a well-established dielectric spectroscopy technique, electrorotation, which has been successfully used in our laboratory to study the alterations of the plasma membrane of cells exposed to various forms of stress. The response to viral proliferation was time dependent as shown by evaluation of the de novo synthesis of viral DNA. This response was paralleled by gradual damage of the membrane evidenced by alteration of the dielectric parameters, specific capacitance and conductance. The electrorotation results show a reduced effect on the dielectric properties of the membrane when infection is carried out in the presence of a natural oil (MEX). In this case a drastic reduction in viral DNA synthesis was also monitored, thus indicating an antiviral action of this product.     

Analysis of Foraging Behaviour of the Whitefly Parasitoid Encarsia formosa on a plant: A Simulation Study

The foraging behaviour of Encarsia formosa was analyzed using a stochastic simulation model of the parasitoid's behaviour. Parasitoids were allowed to search during a day on a tomato plant infested with immatures of the greenhouse whitefly, Trialeurodes vaporariorum. The model simulates searching, host selection, host handling and patch leaving behaviour, and the physiological state of the parasitoid. The outputs of the model are the number of visited leaflets and the number of hosts encountered, parasitized or killed by host feeding. The simulation results agreed well with observations of parasitoids foraging on tomato plants. The number of encounters and ovipositions on the plant increased with host density according to a type II functional response. At a clustered host distribution over leaflets and low host densities, the most important parameters affecting the number of ovipositions were the leaf area, the parasitoid's walking speed and walking activity, the probability of oviposition after encountering a host, the initial egg load and the ratio of search times on both leaf sides. At high densities, the maximum egg load and the giving-up time on a leaflet since latest host encounter were the most essential parameters.     

Deadwood substrate and species-species interactions determine the release of volatile organic compounds by wood-decaying fungi

Wood-decaying fungi in the phylum Basidiomycota play a significant role in the global carbon cycle, as they decompose deadwood effectively. Fungi may compete for utilizable substrate and growth space by producing soluble metabolites and by releasing volatile organic compounds (VOCs). We determined the role of wood substrate (Scots pine or Norway spruce) on the generation of hyphal biomass, secreted metabolites and enzyme activities, wood decomposition rate, and fungal species-species interactions on VOC release. We studied one brown-rot species (Fomitopsis pinicola) and two white-rot species (Phlebia radiata and Trichaptum abietinum) cultivated individually or in combinations. Wood substrate quality influences VOC release by the wood-decaying fungi, with signature differences caused by the decomposition trait (brown rot or white rot) and species-species interactions. VOC release was higher in the cultures of Basidiomycota than in uncolonized sawdust. Fungal biomass, decomposition activity, iron reduction, enzyme activities, oxalate anion content, and oxalic acid production explained VOC release from decaying wood.     

Perception of breath components by the tropical bont tick,Amblyomma variegatum Fabricius (Ixodidae)

Wall-pore olfactory sensilla located in the capsule of Haller's organ on the tarsus of Amblyomma variegatum ticks bear cells responding to vertebrate breath: one of these sensilla contains a CO2-excited receptor and a second sensillum has a CO2-inhibited receptor. Each of these antagonistic CO2-receptors, which display typical phasic-tonic responses, monitors a different CO2-concentration range. The CO2-inhibited receptor is very sensitive to small concentration changes between 0 and ca. 0.2%, but variations of 0.01% around ambient (ca. 0.04%) induce the strongest frequency modulation of this receptor. An increase of just 0.001-0.002% (10-20 ppm) above a zero CO2-level already inhibits this receptor. By contrast, the CO2-excited receptor is not so sensitive to small CO2 shifts around ambient, but best monitors changes in CO2 concentrations above 0.1%. This receptor is characterized by a steep dose-response curve and a fast inactivation even at high CO2-concentrations (greater than 2%). In a wind-tunnel, Amblyomma variegatum is activated from the resting state and attracted by CO2 concentrations of 0.04 to ca. 1%, which corresponds to the sensitivity range of its CO2-receptors. The task of perceiving the whole concentration range to which this tick is attracted would thus appear to be divided between two receptors, one sensitive to small changes around ambient and the other sensitive to the higher concentrations normally encountered when approaching a vertebrate host.     

Proton Transfer Reaction Mass Spectrometry detects rapid changes in volatile metabolite emission by Mycobacterium smegmatis after the addition of specific antimicrobial agents

The metabolic activity of plants, animals or microbes can be monitored by gas headspace analysis. This can be achieved using Proton Transfer Reaction Mass Spectrometry (PTR-MS), a highly sensitive detection method for trace gas analysis. PTR-MS is rapid and can detect metabolic responses on-line as they occur. Here, we study the headspace of actively growing cultures of paired ciprofloxacin sensitive and resistant bacterial strains (Mycobacterium smegmatis in Middlebrook M7H9 liquid media) after the addition of the antibiotics ciprofloxacin and gentamicin in real time. Following the emission patterns of the mycobacteria over time allowed volatile markers specific for the bacterial response to each antibiotic to be detected. A proportion of the measured responses were very rapid, occurring within three hours after the addition of the compounds and varied between isolates with different resistance phenotypes. Specifically, we observed a two fold increase of m73 (unidentified C4 compound) within 10 h after the addition of ciprofloxacin and a threefold increase of m45 (acetaldehyde) within 4 h after the addition of gentamicin as compared to values before the addition. Monitoring the emission of specific volatiles into the culture headspace thus has the potential for rapid drug susceptibility testing. Moreover, these and other differences in the measured responses to the two tested compounds provide evidence that monitoring multiple compounds may also give an indication of the mechanism of action of the compound added.     

The role of ecological availability and host plant characteristics in determining host use by the bogus yucca moth Prodoxus decipiens

1. Specialisation in host plant use is strongly correlated with speciation in many plant‐feeding insects. Specialised taxa, however, could be restricted in host range due to limits in ecological availability of host plant species rather than trade‐offs in using alternate host species. 2. Moths in the genus Prodoxus are extreme specialists on Yucca and speciation is closely tied to host plant shifts. However, many Yucca ranges are allopatric. This study examined whether the bogus yucca moth Prodoxus decipiens is limited in host range because of biogeographic factors or due to differences in the characteristics of host plant species. 3. In a common garden, local P. decipiens moths that use Yucca filamentosa were exposed to individuals of five Yucca species, two that are known hosts of P. decipiens in other parts of its range and three that are used by its sister species, Prodoxus quinquepunctellus. 4. Local moths were attracted to flowers of all Yucca species and females attempted oviposition in the flowering stalks of all species. However, larvae successfully completed development to diapause in only one of the five host plant species. Larval development on non‐natal Yucca species was significantly reduced compared with the local host. 5. The results suggest that differences in host plant characteristics among Yucca species would result in strong natural selection during a host shift. Thus, specialisation in host plant use is probably due to trade‐offs involved with using novel host plant species as well as ecological availability.