Laure Kaiser

Olfactory responses of Trichogramma maidis Pint, et Voeg.: Effects of chemical cues and behavioral plasticity

Oriented responses of Trichogramma maidisPint, et Voeg. to airborne odors were observed in a four-armed olfactometer. Experiments were carried out with odors of host eggs, the sex pheromone of Ostrinia nubilalisHbn, and maize extract, offered singly or in combination, both to naive wasps and to wasps previously exposed to the tested odor during an oviposition experience. The exploratory behavior in the olfactometer was quantified by means of a computer program which performed a space-time analysis of the insect s movements. Whereas the naive wasps did not respond to the odor of the eggs, the synthetic sex pheromone of O. nubilalis,or the maize extract presented singly, they did react to a mixture of these three volatile cues. Prior oviposition in the odor of maize extract or in the combination of odors induced an increased preference toward the conditioning scent. This phenomenon did not occur when the wasps were conditioned to egg odor or sex pheromone alone. These results suggest that females can learn to associate some olfactory cues with the presence of the host. Immediately following the presentation of the combination of odors, a strong attraction of experienced wasps occurred; it decreased as the experiment progressed and finally reached the level presented by naive insects. Adult conditioning to the combination of odors also resulted in reduced variability in the behavioral responses.

Parametric and genetic analysis of Drosophila appetitive long-term memory and sugar motivation.

Distinct forms of memory can be highlighted using different training protocols. In Drosophila olfactory aversive learning, one conditioning session triggers memory formation independently of protein synthesis, while five spaced conditioning sessions lead to the formation of long‐term memory (LTM), a long‐lasting memory dependent on de novo protein synthesis. In contrast, one session of odour–sugar association appeared sufficient for the fly to form LTM. We designed and tuned an apparatus that facilitates repeated discriminative conditioning by alternate presentations of two odours, one being associated with sugar, as well as a new paradigm to test sugar responsiveness (SR). Our results show that both SR and short‐term memory (STM) scores increase with starvation length before conditioning. The protein dependency of appetitive LTM is independent of the repetition and the spacing of training sessions, on the starvation duration and on the strength of the unconditioned stimulus. In contrast to a recent report, our test measures an abnormal SR of radish mutant flies, which might initiate their STM and LTM phenotypes. In addition, our work shows that crammer and tequila mutants, which are deficient for aversive LTM, present both an SR and an appetitive STM defect. Using the MB247‐P[switch] system, we further show that tequila is required in the adult mushroom bodies for normal sugar motivation.

Odor learning byLeptopilina boulardi, a specialist parasitoid (Hymenoptera: Eucoilidae)

We studied odor learning in Leptopilina boulardi,a specialist larval parasitoid of Drosophila melanogaster.The behavioral responses of differently experienced females to an artificial odor (Must de Cartier, Paris) were analyzed using a fourarmed airflow olfactometer. The responses of females with an oviposition experience in the presence of the perfume were compared with those of four control groups. As controls we used naive females, females with an oviposition experience in the absence of odor, females which had been previously exposed to perfume but without an oviposition experience, and females with an oviposition experience which also had been exposed to perfume but not at the same time. The results demonstrate that a specialist such as L. boulardican learn very well to respond to an artificial odor by associating this odor with a reward, i.e., an oviposition. The four control groups responded more or less in a similar way.

Social Facilitation of Long-Lasting Memory Retrieval in Drosophila

Recent studies demonstrate that social interactions can have a profound influence on Drosophila melanogaster behavior and cuticular pheromone patterns. Olfactory memory performance has mostly been investigated in groups, and previous studies have reported that grouped flies do not interact with each other and behave in the same way as individual flies during short-term memory retrieval. However, the influence of social effects on the two known forms of Drosophila long-lasting associative memory, anesthesia-resistant memory (ARM) and long-term memory (LTM), has never been reported. We show here that ARM is displayed by individual flies but is socially facilitated; flies trained for ARM interact within a group to improve their conditioned performance. In contrast, testing shows LTM improvement in individual flies rather than in a group. We show that the social facilitation of ARM during group testing is independent of the social context of training and does not involve nonspecific aggregation. Furthermore, we demonstrate that social interactions facilitate ARM retrieval. We also show that social interactions necessary for this facilitation are specifically generated by trained flies: when single flies trained for ARM are mixed with groups of naive flies, they display poor retrieval, whereas mixing with groups trained either for ARM or LTM enhances performance.

The influence of plant species on attraction and host acceptance inCotesia glomerata (Hymenoptera: Braconidae)

Females of the larval parasitoidCotesia glomerata (L.) use plant-associated cues to locate their lepidopteran host,Pieris rapae L. In this study we investigated the influence of four host plant species,Brassica oleracea var.acephala (‘Vates’ kale),Tropaeolum majus (nasturtium),Lunaria annua (honesty), andCleome spinosa (spider flower), on two components of the host selection process inC. glomerata, namely, attraction and host acceptance. Choice tests in a flight tunnel showed that parasitoids were attracted to some host plant species more than to others in the absence of host larvae.B. oleracea was the most attractive plant species, followed byL. annua, T. majus, andC. spinosa. In previous studies it was shown thatB. oleracea carries highly suitable hosts forC. glomerata and that, in the field, parasitization rates on this plant were the highest. When host larvae were reared on the four host plant species and then transferred to a common substrate (B. oleracea var.capitata, cabbage), plant species that had served as diet for the hosts did not have a significant effect on acceptance for parasitization. Thus, parasitoids were attracted to host plant species differentially, but they did not discriminate among host larvae based on the dietary history of their hosts. ForC. glomerata, it appears that phytochemistry mediates host selection more by influencing parasitoid attraction than it does by affecting host acceptance.

Parasitism of canola aphids in France in autumn

In light of a recent increase in the aphid populations on young canola (Brassica napus) in autumn in Northwestern Europe, we carried out a survey of their parasitoid species during this season. The study was done in France from 1998 to 2001 using different sampling methods. Results highlighted the presence of two main species of Aphidiinae, Aphidius matricariae and Diaeretiella rapae (and to a lesser extent Aphidius ervi) on Myzus persicae, and of D. rapae on Brevicoryne brassicae. Nine other Aphidiinae species were found occasionally as well as some Aphelinidae parasitoids and hyperparasitoids. There was no difference in parasitoid species pattern between fields in Northern and Southern France. The principal parasitoid species found in the fields were reared in the laboratory to confirm their ability to develop on the canola aphids. Aphidius matricariae and D. rapae were reared successfully on M. persicae, but a low parasitism rate was obtained for A. ervi on this aphid. This study showed that A. matricariae and D. rapae could develop on aphids on canola and that they are present naturally in canola in autumn. However, the parasitism rate is low in autumn, so the options could be the use of these parasitoids in augmentative release biological control programs or in an IPM project on canola fields during this time of the year.

Induction of odor preference in a specialist insect parasitoid

This paper deals with the effect of associative and nonassociative odor conditioning on odor preference in a specialist larval parasitic wasp ofDrosophila melanogaster, Leptopilina boulardi. Differently experienced females were offered a choice between banana and strawberry odors in a four-armed olfactometer. Wasps that were allowed to oviposit in an odorless airflow before testing exhibited no significant response to the odors, whereas females with an oviposition experience in the presence of an odor clearly preferred the scent that they had experienced. Furthermore, exposure to strawberry odor after oviposition could also induce a preference for strawberry or banana odor, depending on the concentrations of both scents used for conditioning and testing. This effect was independent of the time elapsed between oviposition and strawberry exposure, suggesting that it was caused by nonassociative conditioning and not by backward associative learning. Exposure to banana odor after oviposition had no effect on females’ responses. Variations in odor concentrations also influenced the expression of associative learning, although to a much lesser extent.

Ongoing ecological speciation in Cotesia sesamiae, a biological control agent of cereal stem borers

To develop efficient and safe biological control, we need to reliably identify natural enemy species, determine their host range, and understand the mechanisms that drive host range evolution. We investigated these points in Cotesia sesamiae, an African parasitic wasp of cereal stem borers. Phylogenetic analyses of 74 individual wasps, based on six mitochondrial and nuclear genes, revealed three lineages. We then investigated the ecological status (host plant and host insect ranges in the field, and host insect suitability tests) and the biological status (cross‐mating tests) of the three lineages. We found that one highly supported lineage showed all the hallmarks of a cryptic species. It is associated with one host insect, Sesamia nonagrioides, and is reproductively isolated from the other two lineages by pre‐ and postmating barriers. The other two lineages had a more variable phylogenetic support, depending on the set of genes; they exhibited an overlapping and diversified range of host species and are not reproductively isolated from one another. We discuss the ecological conditions and mechanisms that likely generated this ongoing speciation and the relevance of this new specialist taxon in the genus Cotesia for biological control.

Computer analysis of the exploratory behavior of insects and mites in an olfactometer.

A method of quantification of the exploratory behavior of small animals stimulated by an odorant in a four-choice olfactometer, taking into account the interindividual variability of responses, was developed: individual tracks were time sampled according to the animals walking speed and its positions were recorded according to the X-Y coordinates of the grid set underneath the device, the mesh of the grid suiting the animals body size. A software, written in BASIC APPLESOFT on an APPLE IIe computer, allowed us to analyze the coordinates either of a single individual or of an experimental sample, leading to: a) the quantification of the insect distribution all over the experimental chamber, expressed in a table numbered according to the grid, where the percentage of position per square either for a given time fraction or the total observation period were reported, b) a graphic representation of the data according to several levels of greys, expressing the frequentation for each square for a given duration of observation. An analysis per time fraction allowed the chronological setup of events to appreciate. c) The collection of the positions among each flow field of the olfactometer for each individual of the experimental sample, for a given duration, was translated as the percentage of time spent in each flow field. Data files gathered these percentages for further statistical treatments. This computer method, which requires little equipment and appears to be easily adaptable to the study of biological models of various size and speed such as honeybees, trichogrammas and varroas mites, is a powerful tool for behavioral studies of small organisms tested in restricted areas.

Behavioral characterization of individual olfactory memory retrieval in Drosophila melanogaster

Memory performance depends not only on effective learning and storage of information, but also on its efficient retrieval. In Drosophila, aversive olfactory conditioning generates qualitatively different forms of memory depending on the number and spacing of conditioning trials. However, it is not known how these differences are reflected at the retrieval level, in the behavior of individual flies during testing. We analyzed conditioned behaviors after one conditioning trial and after massed and spaced repeated trials. The single conditioning produces an early memory that was tested at 1.5 h. Tested at 24 h after training, the spaced and the massed protocols generate two different forms of consolidated memory, dependent, or independent of de novo protein-synthesis. We found clearly distinct patterns of locomotor activity in flies trained with either spaced or massed conditioning protocols. Spaced-trained flies exhibited immediate and dynamic choices between punished and unpunished odors during the test, whereas massed-trained flies made a delayed choice and showed earlier disappearance of the conditioned response. Flies trained with single and spaced trials responded to the punished odor by decreasing their resting time, but not massed-trained flies. These findings demonstrate that genetically and pharmacologically distinct forms of memory drive characteristically different forms of locomotor behavior during retrieval, and they may shed light on our previous observation that memory retrieval in massed-trained flies is socially facilitated. Social interactions would enhance exploratory activity, and then reduce the latency of their conditioned choice and delay its extinction.