Heritable differences in the response of the braconid waspMicroplitis croceipes to volatile allelochemicals

The heritable nature of differential responses by Microplitis croceipes(Cresson) (Hymenoptera: Braconidae) to airborne allelochemicals was investigated. Four isofemale lines were tested for three generations in a flight tunnel. Flight response was found to be strongly dependent on the familial origin, with two highly responsive lines being clearly differentiated from two less responsive ones. Each isofemale line could also be recognized by typical behaviors that suggested that M. croceipesfemales may inherit independent characters of sensory acuity to plant and to host chemicals and of learning ability.     

Behavioral time budget and periodicity exhibited byMicroplitis croceipes in field cages withHeliothis virescens on spring host plants

Microplitis croceipes (Cresson) females were observed between 06:00 and 20:00 hours in cages containing spring host plants infested withHeliothis virescens (F.), tobacco budworm, larvae, Cages containedGeranium dissectum L. andTrifolium incarnatum L. in plantings of equal area, each infested with 50H. virescens larvae. A time budget for the various behaviors exhibited by the wasps was obtained. In order of decreasing time spent,M. croceipes females were observed to exhibit standing still, searching on plant surface, preening, oriented flight, general flight, and walking. The proportion of time spent preening increased over the day. Oriented flight, general flight, searching, and walking peaked in the morning between 09:00 and 12:00 hours. Oriented flight, general flight, and walking showed minor increases in the afternoon.Microplitis croceipes females spent significantly more time preening, orienting to, and walking and searching on clover than on geranium. However, there was no difference in time spent standing still on geranium or clover. Significance of these findings for development of effective augmentative release strategies is discussed.     

The role of herbivore‐ and plant‐related experiences in intraspecific host preference of a relatively specialized parasitoid

Parasitoids use odor cues from infested plants and herbivore hosts to locate their hosts. Specialist parasitoids of generalist herbivores are predicted to rely more on herbivorederived cues than plant-derived cues. Microplitis croceipes (Cresson)(Hymenoptera: Braconidae) is a relatively specialized larval endoparasitoid of Heliothis virescens (F.)(Lepidoptera: Noctuidae), which is a generalist herbivore on several crops including cotton and soybean. Using M. croceipes/H. virescens as a model system, we tested the following predictions about specialist parasitoids of generalist herbivores:(i) naive parasitoids will show innate responses to herbivore-emitted kairomones, regardless of host plant identity and (ii) herbivore-related experience will have a greater influence on intraspecific oviposition preference than plant-related experience. Inexperienced (naive) female M. croceipes did not discriminate between cotton-fed and soybean-fed H. virescens in oviposition choice tests, supporting our first prediction. Oviposition experience alone with either host group influenced subsequent oviposition preference while experience with infested plants alone did not elicit preference in M. croceipes, supporting our second prediction. Furthermore, associative learning of oviposition with host-damaged plants facilitated host location. I terestingly, naive parasitoids attacked more soybeathan cotton-fed host larvae in two-choice tests when a background of host-infested cotton odor was supplied, and vice versa. This suggests that plant volatiles may have created an olfactory contrast effect. We discussed ecological significance of the results and concluded that both plant- and herbivore-related experiences play important role in parasitoid host foraging.     

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     

Disruptive Sublethal Effects of Insecticides on Biological Control: Altered Foraging Ability and Life Span of a Parasitoid after Feeding on Extrafloral Nectar of Cotton Treated with Systemic Insecticides

Predictions in integrated pest management on the compatibility of an insecticide with biological control often are based on incomplete screening tests. While measuring levels of mortality from direct insecticide exposure is a very common screening method, possible sublethal effects as a result of either direct or indirect insecticide exposure remain relatively unknown. The impact of sublethal effects on the success of biological control can be as deleterious as mortality. Here, we report the reduced host foraging ability and longevity of the parasitoid Microplitis croceipes Cresson (Hymenoptera: Braconidae) after feeding on extrafloral nectar from cotton (Gossypium hirsutum L., Malvaceae) plants that were treated with systemic insecticides. The insecticides used in this study are regularly applied in cotton-growing areas in the United States. For all tested insecticides, longevity of M. croceipes females that fed on nectar from cotton was affected for at least 10 days after plants were treated with insecticides. Moreover, the parasitoid's host foraging ability was severely affected for periods ranging from 2 days (imidacloprid) to 18 days (aldicarb) after insecticide application. The consequences of these sublethal effects on the success of biological control are discussed.     

Toxic and behavioral effects of selected insecticides on theHeliothis parasitoidMicroplitis croceipes

Toxic and behavior-modifying actions of several formulated insecticides were determined forMicroplitis croceipes (Cresson), a braconid parasitoid ofHeliothis spp. Exposure of adult parasitoids to cotton plants sprayed at recommended field rates with a pyrethroid/formamidine mixture (fenvalerate/chlordimeform) resulted in significantly higher mortality rates (10.4 to 22.6 percent) than in controls. Exposure to the carbamate thiodicarb resulted in similar rates of mortality. Only methomyl, a type of carbamate different from thiodicarb, caused mortality significantly higher than all other treatments, ranging to about 70%. Flight activity was measured by attraction to cotton in a laboratory wind tunnel bioassay. Females sprayed directly with a fenvalerate/chlordimeform mixture had significantly decreased flight activity up to 20 h post-treatment. Alternatively, attraction to cotton sprayed with either the fenvalerate/chlordimeform mixture or with methomyl to unsprayed females was significantly decreased, compared to plants sprayed with water only. These results suggest that the actions of insecticides, other than those of direct toxicity, may be important on beneficial parasitoids. Behaviors, such as flight activity and foraging, may be altered by even relatively non-toxic insecticides, thus potentially modifying the effectiveness of natural enemies. This article reports the results of research only. Mention of a proprietary product does not constitute endorsement or a recommendation for its use by the U.S. Department of Agriculture.     

Flight responses in a wind tunnel of the parasitoid Microplitis croceipes to three spring and three summer host plants of Heliothis spp.

Flight responses of the parasitoid, Microplitis croceipes (Cresson) (Hymenoptera: Braconidae) to two phenological stages of spring host plants (crimson clover, Trifolium incarnatum L., hairy vetch, Vicia villosa Corbiere, and cutleaf geranium, Geranium dissectum L.) and summer host plants (cotton, Gossypium hirsutum L., sorghum, Sorghum bicolor L., and soybean Glycine max (L.) Merr.) of Heliothis virescens (F.), were measured in a dual choice situation in a wind tunnel. Preflowering clover was more attractive than preflowering geranium, followed by preflowering vetch. Clover was preferred over geranium in flowering stages as well. No within species differences were detected for the two phenological stages. Preflowering and flowering cotton and soybean were preferred over preflowering and flowering sorghum. The findings could be important in pre-determining release sites for M. croceipes in the field.     

Responses of Microplitis croceipes to host and nonhost plants of Heliothis virescens in a wind tunnel

The relative attractiveness of velvet leaf (Abutilon theophrasti Medicus), cotton (Gossypium hirsutum L.) (host plants) and groundcherry (Physalis angulata L.) (nonhost plant), and cotton plants with or without nectaries and with or without glands to Microplitis croceipes Cresson (Hymenoptera: Braconidae) was determined in a wind tunnel. Female parasitoids flew significantly more to glandless than to glanded cotton; response to nectaried and nectariless cotton was similar. Velvet leaf and cotton were favored significantly over groundcherry; parasitoids being equally responsive to both host plant species. Addition of larval frass alone or in combination with host larvae significantly improved the attractivity of the nonhost plant (groundcherry) to the parasitoids. There was no difference in attractiveness of groundcherry terminals with or without host larvae. Parasitoid search time was significantly increased with addition of larval frass. In the presence of cotton, however, kairomone-treated groundcherry remained unattractive.

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Résumé L'influence des nectaires et des glandes du coton, et l'attractivité des feuilles veloutées du coton (plante hôte) et de Physalis angulata sur Microplitis croceipes, ont été déterminées dans des expériences avec tunnel à vent. Les résultats ont montré que les parasitoïdes femelles sont significativement plus attirés par les cotons sans glandes que par les cotons glanduleux, tandis que la présence ou l'absence de nectaires ne modifie pas l'attractivité du coton. Elles réagissent de la même façon aux feuilles veloutées. P. angulata traité avec des crottes de chenilles, présenté seul ou en combinaison avec du coton, attire plus de femelles que des pousses de P. angulata non traitées. L'addition de chenilles hôtes seules n'améliore pas significativement l'attractivité de la plante. P. angulata traité avec des kairomones est demeuré inattractif quand il était proposé en même temps que du coton.

     

Learning of host searching cues by the larval parasitoid Microplitis croceipes

We conducted laboratory experiments using a wind tunnel to determine the effects of prior experience on the learning and retention of learned responses in the larval parasitoid Microplitis croceipes (Cresson) (Hymenoptera: Braconidae). Although most females that had either antennated host frass or oviposited in a host in the presence of vanilla odor, made oriented flight to the odor from downwind in the wind tunnel at 30 min after experience, only those that had oviposited in a host with the odor responded to the odor at 24 h after experience. Females that had oviposited in the non‐natural host beet armyworm (BAW) larva with or without the odor did not respond to the odor at 30 min after experience. These results indicate that an oviposition in the host in the presence of odors strongly affects associative learning and the persistence of learned response to the odors. When females were allowed to antennate host frass in the presence of vanilla and to subsequently oviposit in a host within an interval of 5 min or less, their learned response to vanilla also persisted for 24 h. Similarly, when females were conditioned to link vanilla with host frass and then allowed to make an ovipositor contact with host hemolymph, their learned response persisted for 24 h. However, antennal contact with hemolymph after such conditioning of vanilla with host frass did not improve the persistence of learned response to the odor. These results indicate that ovipositor contact with host hemolymph during oviposition is partially responsible for an increased retention of learned response. Females responded to vanilla 48% of the time at 30 min after antennating host frass without the odor, but their response to the odor significantly decreased after oviposition in a BAW larva subsequent to the antennation of host frass. This result indicates that oviposition in a BAW larva decreases subsequent response to general odors. Based on the results, we discuss the foraging behavior of M. croceipes dependent on learning and subsequent experiences.     

Nitrogen and water affect direct and indirect plant systemic induced defense in cotton

Plants have direct and indirect constitutively produced and inducible defenses against herbivores and pathogens, which can substantially aid in their ability to defend themselves. However, very little is known about the influence of agronomic factors on such defenses. Here, we tested the effects of nitrogen levels and water availability on the ability of cotton plants to deter feeding by Spodoptera exigua through induction of anti-feedants, and to attract Microplitis croceipes through systemic induction of volatile emission. Cotton plants were grown with various nitrogen levels and were either exposed to water stress or normal water before being exposed to S. exigua for 48 h for induction of defenses. Dual choices of various nitrogen and water treatments were provided to M. croceipes in flight tunnel bioassays. Dual choices of leaf tissue from the various nitrogen and water treatments were provided to S. exigua larvae. Both water stress and nitrogen levels under and over the recommended levels increased leaf tissue consumption and decreased attraction of M. croceipes to the plants. Analyses of induced volatiles released from herbivore damaged plants indicate that their concentrations differ among the nitrogen levels tested with plants receiving no nitrogen or twice the recommended dose having amounts much lower than plants receiving the recommended dose. Because both direct and indirect plant defense mechanisms are negatively affected by improper nitrogen and insufficient water, we argue that these factors should be considered for a better natural control of pests in cotton and most probably in other crops.     

Host recognition by the specialist endoparasitoidMicroplitis croceipes (Hymenoptera: Braconidae): Role of host- and plant-related volatiles

The specialist parasitoidMicroplitis croceipes Cresson can parasitize only noctuid larvae in the generaHelicoverpa andHeliothis. To be successful in their search for hosts, the ability to distinguish hosts from nonhosts feeding on the same plant is beneficial. In flight tunnel experiments, we found that prior to landing on the odor sourceM. croceipes were able to distinguish volatiles released from frass of host larvae(Helicoverpa zea Boddie) and nonhost larvae (Spodoptera exigua Hübner andSpodoptera frugiperda J. E. Smith) fed on cotton. However, an initial contact experience with frass of cotton-fed host larvae appeared to be critical for this ability. Wasps that had antennated frass of host larvae fed pinto bean diet were equally attracted to frass of host and nonhost larvae fed on pinto bean diet. In short-range walking experiments, wasps located cotton-fed host larvae faster than diet-fed larvae, regardless of their experience. Wasps that had antennated frass of cotton-fed host larvae were less attracted to cotton-fed nonhost larvae, compared to host larvae, and preferred to sting host larvae. Plant-related volatiles in host frass and larvae appear to play a major role in the successful location of host larvae.     

Influence of handling and conditioning protocol on learning and memory of Microplitis croceipes

Microplitis croceipes (Cresson) (Hymenoptera: Braconidae) learns odors in association with both hosts and food. The food‐associated ‘seeking’ behavior of M. croceipes was investigated under various training protocols utilizing the conditioning odor, 3‐octanone. We investigated the effects of odor training, or its lack, training duration, training frequency, time elapsed after training, wasp hunger state, and training reinforcement, on the food‐seeking responses of M. croceipes females. We found that odor‐trained females show strong food seeking responses, whereas non‐odor‐trained females do not respond to the odor, and that a single 10 s association with the odor whilst feeding on sugar water subsequently conditioned the wasps to exhibiting significant responses to it. Increases in training time to more than 10 s did not improve their responses. Repetition of the food–odor associations increased a wasp's recall, as well as its response over time, compared to a single exposure. Repeated exposure to the learned odor in the absence of a food reward decreased the responses of less hungry individuals. However, the level of response increased significantly following a single reinforcement with the food–odor association. Understanding the factors that influence learning in parasitoids can enhance our ability to predict their foraging behavior, and opens up avenues for the development of effective biological detectors.     

Conditioning of appetitive behavior in the Hymenopteran parasitoid Microplitis croceipes

Work on insect learning has made ample use of conditioned behaviors in single learning tasks. Parasitic wasps are particularly suited to study more complex learning processes, as they alternate between host searching and foraging for sugar sources. We here describe a set of behaviors that unambiguously represents sugar feeding in the Hymenopteran parasitoid Microplitis croceipes (Cresson) (Hymenoptera: Braconidae). Furthermore, we demonstrate that this parasitoid can be conditioned to exhibit these behaviors in response to odor stimuli previously associated with feeding. In conjunction with a previously described ovipositor probing response in this species, our findings constitute the first instance in which multiple conditioned behaviors representing feeding and reproduction have been identified for one insect species. This provides a valuable tool for the study of multiple task learning.     

Analyses of Two Parasitoids with Convergent Foraging Strategies

We compared the foraging strategies of two key braconid endoparasitoids of the tobacco budworm (Heliothis virescens Fab.), Cardiochiles nigriceps Vier. and Microplitis croceipes Cresson, that differ in host and habitat range but otherwise share comparable, overlapping niches. The most important host-location cues by far for both species were materials associated with damaged plants. Both species demonstrated a significant preference for volatiles released from plants damaged by H. virescens larvae over those released from undamaged tobacco and cotton plants. In choice experiments with damaged tobacco versus cotton, M. croceipes showed a significant preference for cotton plants. In contrast, C. nigriceps preferred damaged tobacco plants. Plant compounds provoked a strong response even when released from systemically induced plants (from which damaged leaves, host, and host by-products were removed). C. nigriceps appears to have a much keener ability to locate hosts over long distances than M. croceipes. This observation may be related to the highly specialized nature of this parasitoid. The possible adaptive significance of the foraging behaviors of these two parasitoids is discussed.     

Intrinsic and extrinsic competitive interactions between two larval parasitoids of Heliothis virescens

1. Competition between parasitoid species may be a key factor in the community dynamics of plant–herbivore-parasitoid systems and is an important consideration in the selection and management of effective biological control agents. 2. Interspecific competition can occur between adult parasitoids searching for hosts (extrinsic competition) and between multiple parasitoid larvae developing within a single host individual (intrinsic competition). A model system comprising the lepidopteran pest Heliothis virescens and two key hymenopteran endoparasitoids, Microplitis croceipes and Cardiochiles nigriceps, was employed to explore parasitoid host-location strategies and the consequences of intrinsic and extrinsic competitive interactions between parasitoid species. 3. The less specialised of the two parasitoids, M. croceipes, was found to have a shorter hatching time and to dominate intrinsic competition, except when its oviposition followed that of the more specialised parasitoid, C. nigriceps, by 16 h or more. This interval corresponded to the differential in hatching time between the two species. 4. Cardiochiles nigriceps, however, displayed superior host-searching efficiency that may compensate for its disadvantage in intrinsic competition. This parasitoid was more effective at detecting host infestation sites via airborne odours and at locating and attacking early instar host larvae than was M. croceipes.     

Influence of an endophytic fungus on host plant selection by a polyphagous moth via volatile spectrum changes

Recently the role of micro-organisms as mediators of plant-herbivore interactions has been increasingly acknowledged in ecological research. We investigated the interaction between an unspecialized root fungal endophyte (Acremonium strictum) and the polyphagous moth Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), in greenhouse and laboratory bioassays. Specifically we examined in tomato (Lycopersicon esculentum Mill.) the systemic effects of the endophytic fungus on the host selection behaviour of female moths for oviposition and the volatile profiles of host plants to understand the mechanisms acting in this multi-trophic model system. Both laboratory and field strains of H. armigera moths oviposited more on leaves of A. strictum inoculated plants as compared to endophyte-free plants, both in free flight cages and in tethered moth laboratory experiments; the moth’s preferences were significant between 10 and 18 days after inoculation. The analysis of volatile profiles showed strong quantitative differences between treatments. Endophyte inoculated plants emitted diverse terpenes and sesquiterpenes at significantly lower amounts as compared to endophyte free-plants, except for α-terpinene, which did not differ between the treatments, and trans-β-caryophyllene, which was emitted in significantly higher amounts on inoculated plants. β-Thujene and α-phellandrene accounted for 73.3 and 12.0% of total amounts of volatiles emitted from endophyte-free and inoculated plants, respectively. Our findings demonstrate that A. strictum is able to systemically influence the host selection of H. armigera moths for oviposition; conceivably mediated by the induced changes in volatile emissions (and probably additional biochemical parameters of the host plants, which have not been analysed so far). We argue for a more detailed assessment of micro-organisms invisibly colonizing plants, when studying plant-herbivore or multitrophic interactions.     

Discrimination of previously searched,host-free sites byMicroplitis croceipes (Hymenoptera: Braconidae)

The ability of free- ranging parasitoids to discriminate between previously visited and unvisited sites containing host kairomone (caterpillar frass) but not hosts was tested. Females of Microplitis croceipes,a host specialist and plant generalist larval parasitoid of Helicoverpa (Heliothis) zea,were allowed to fly freely in a simulated plant patch in a flight chamber. Wasps spent less time searching frass sites previously searched by themselves or by conspecifics than unsearched frass sites. In addition to chemical marking, spatial memory of visual cues was implicated as a mechanism for discriminating against self-visited, host-free sites.     

Effect of Adult Experience on in-Flight Orientation to Plant and Plant–Host Complex Volatiles inAphidius erviHaliday (Hymenoptera, Braconidae)

The effect of adult experience on in-flight orientation to plant–host complex volatiles byAphidius erviHaliday was studied in a wind tunnel bioassay, usingAcyrthosiphon pisum(Harris), maintained on broad bean plants (Vicia faba) as host. A short oviposition experience (15 s) on the plant–host complex (PHC) was sufficient to induce a drastic decrease of flight propensity and stimulated a foraging behavior characterized by intense walking activity. However, flight activity resumed to normal levels 1 h after the oviposition experience on the PHC occurred. For parasitoids conditioned on the PHC for at least 1 min the recorded proportion making oriented flights to the PHC was significantly higher than that for naive females. In contrast, oviposition experience in the absence of plant material did not influence theA. erviflight response. Oviposition attempts on aphid dummies without egg release did not reduce flight activity. WhenA. ervifemales were exposed to glass beads coated withAc. pisumcornicle secretion, a priming effect was observed, resulting, compared with naive females, in a significantly higher rate of oriented flights to the PHC. In contrast, oviposition attempts visually induced by colored aphid dummies did not influence flight behavior. A strong reaction to volatile cues from uninfested plants was induced by oviposition experience on newly infested broad bean plants. This appears to be a case of associative learning. In fact, uninfested broad bean plants are basically unattractive to naiveA. ervifemales. The results demonstrate that adult experience has a considerable influence onA. ervibehavior and may have important implications for biological control of natural pest aphid populations.     

Influence of maternal age and host deprivation on egg production and parasitization byMicroplitis croceipes [Hym.: Braconidae]

The effect of maternal age and hostHeliothis virescens (F.) deprivation on egg production and allocation byMicroplitis croceipes (Cresson) was determined in the laboratory on 0.5, 1.0, 3.0, 6.0, 9.0, and 12.0 days old parasitoids. In host-deprived parasitoids, newly-emerged (0.5-day old) females produced significantly fewer eggs than older ones; egg production peaked 3 to 6 days after emergence and declined thereafter. However, the number of eggs found in undeprived parasitoids at various ages suggested a cyclical pattern of egg production. The rate of parasitization by host-deprivedM. croceipes was higher in 1.0 to 6.0 days old than in newly-emerged or older females and was correlated strongly with egg production (r=0.93). The first observation on premature larval hatching inM. croceipes was documented.      

Comparative flight behavior of parasitoids Campoletis sonorensis and Microplitis croceipes

Flight behavior of parasitoids Microsplitis croceipes Cresson (Braconidae) and Campoletis sonorensis (Cameron) (Ichneumonidae) was examined in a wind tunnel. Both species exhibited odor directed, oriented flights to cotton, although flight frequency was related to variation in wind velocity or differential cotton volatile emission with changes in wind velocity, or both. Flight frequency at constant wind velocity was affected by illumination. Microplitis croceipes was attracted to the wind borne odor of Heliothis virescens (F.) frass and larvae, whereas C. sonorensis was not. Possible interaction of the wind borne odors from the habitat and host is suggested for parasitoids.

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Résumé Le comportement de vol de M. croceipes Cres. (Hym. Braconidae) et de C. sonorensis Cam. (Hym. Icheumonidae) a été étudié en olfactométrie dans un tunnel à vent. Les femelles des deux espèces, et non les mâles, ont un vol orienté par l'odeur des feuilles fraîches de coton (Gossypium hirsutum L.). La fréquence d'envol est liée aux changements de vitesse du vent, aux variations d'émissions, par le cotonnier, des substances volatiles, en fonction de la vitesse du vent, ou aux deux. La fréquence d'envol à vitesse de vent constante dépend de l'éclairage. La fréquence d'envol de C. sonorensis augmente avec des intensités lumineuses relativement fortes et diminue avec des intensités relativement faibles. L'inverse est observé avec M. croceipes. Le vent chargé d'odeurs de larves ou d'extréments d'Heliothis virescens F., attire M. croceipes et non C. sonorensis. Nous suggérons une éventuelle interaction des odeurs portées par le vent et provenant de l'habitat et de l'hôte.