Résultat de la recherche
Tritrophic interactions on cultivated maize and its wild ancestor "teosinte"
Modern maize plants (Zea mays ssp. mays, Poaceae) are characterized by large cobs that contain juicy grains, although they have not always had these characteristics. Approximately 9000 years ago, maize was domesticated from teosinte (Z. mays ssp. parviglumis), its closest wild ancestor, which produces much less and much smaller seeds. Teosinte still grows in the wild in Mexico, while maize is produced all over the world. Continuous selection for improved yield and quality has had a cost for the plant in terms of the loss or alteration of other potentially useful traits, such as resistance to pathogens and herbivorous insects. This thesis focuses on the resistance of maize and teosinte against insect pests, in particular with respect to the emission of herbivore-induced volatiles. This feature is considered an indirect defense trait, as the volatiles can betray the presence of prey or hosts to predators and parasitoids, natural enemies of herbivorous insects. Among these natural enemies are parasitoid wasps, of which females have an ovipositor with which they can lay single or multiple eggs in individual hosts. When the wasp larvae develop inside the host they will eventually kill it, potentially benefitting the plant.
At first, we assessed in nature which insects occur on teosinte. Fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), is reported as an important maize pest in the Americas and is frequently attacked by many species of parasitoids. However, little is known about the presence of this herbivore and its associated parasitoids on teosinte.
In a laboratory setting, we then assessed whether maize and teosinte emit a similar blend of volatiles when induced by lepidopteran herbivores. We also evaluated the attractiveness of the odor blends to females of two species of parasitoid wasps. Although the odor blends appeared similar upon gas chromatography analysis, the foraging parasitoids responded differentially to them. We obtained similar results when testing real plants and extracts of collected odors, indicating that these extracts can be used to identify the key compounds that are responsible for parasitoid attraction.
There is surprisingly little field evidence for fitness benefits for plants due to the action of natural enemies. This is still an important point in the discussion on the possible indirect defense role of inducible plant volatiles, especially in the case of parasitoids that, unlike predators, do not directly kill their hosts. Therefore, we evaluated how parasitoid wasps can affect plant performance in a semi-natural setting in Mexico. The presence of parasitoid wasps in field tents containing teosinte plants and fall armyworm significantly reduced herbivore damage, which, for the smallest plants, resulted in a reduction in plant mortality. These findings support the notion that plants may benefit from the presence of parasitoids and may help to resolve the current debate on the defensive function of herbivore-induced volatiles.
To further explore the importance of volatiles for the attraction of parasitoids under field conditions, we studied maize lox10 mutants, impaired in the biosynthesis of green leaf volatiles (GLVs). These volatiles, responsible for the smell of cut grass, are commonly emitted by plants when wounded or attacked by herbivores. Previously, laboratory studies have shown that GLVs can be used as foraging cues by predators and parasitoids. However, our results imply that GLVs are not of key importance for parasitoid attraction in the field.
Collectively, these studies provide novel insights into the importance of herbivore-induced volatiles for the attraction of parasitoid wasps, and their indirect importance for plant growth and survival. We identified differences between maize and its wild ancestor, teosinte, with respect to the attraction of parasitoids that could possibly be exploited for ecologically sound methods to better protect maize against insect pests.
Fungal Infection Reduces Herbivore-Induced Plant Volatiles of Maize but does not Affect Naïve Parasitoids
Plants attacked by insects release volatile compounds that attract the herbivores' natural enemies. This so-called indirect defense is plastic and may be affected by an array of biotic and abiotic factors. We investigated the effect of fungal infection as a biotic stress agent on the emission of herbivore-induced volatiles and the possible consequences for the attraction of two parasitoid species. Maize seedlings that were simultaneously attacked by the fungus Setosphaeria turcica and larvae of Spodoptera littoralis emitted a blend of volatiles that was qualitatively similar to the blend emitted by maize that was damaged by only the herbivore, but there was a clear quantitative difference. When simultaneously challenged by fungus and herbivore, the maize plants emitted in total 47% less of the volatiles. Emissions of green leaf volatiles were unaffected. In a six-arm olfactometer, the parasitoids Cotesia marginiventris and Microplitis rufiventris responded equally well to odors of herbivore-damaged and fungus- and herbivore-damaged maize plants. Healthy and fungus-infected plants were not attractive. An additional experiment showed that the performance of S. littoralis caterpillars was not affected by the presence of the pathogen, nor was there an effect on larvae of M. rufiventris developing inside the caterpillars. Our results confirm previous indications that naïve wasps may respond primarily to the green leaf volatiles.
Strong Attraction of the Parasitoid Cotesia marginiventris Towards Minor Volatile Compounds of Maize
Plants infested with herbivorous arthropods emit complex blends of volatile compounds, which are used by several natural enemies as foraging cues. Despite detailed knowledge on the composition and amount of the emitted volatiles in many plant-herbivore systems, it remains largely unknown which compounds are essential for the attraction of natural enemies. In this study, we used a combination of different fractionation methods and olfactometer bioassays in order to examine the attractiveness of different compositions of volatile blends to females of the parasitoid Cotesia marginiventris. In a first step, we passed a volatile blend emitted by Spodoptera littoralis infested maize seedlings over a silica-containing filter tube and subsequently desorbed the volatiles that were retained by the silica filter (silica extract). The volatiles that broke through the silica filter were collected on and subsequently desorbed from a SuperQ filter (breakthrough). The silica extract was highly attractive to the wasps, whereas the breakthrough volatiles were not attractive. The silica extract was even more attractive than the extract that contained all herbivore-induced maize volatiles. Subsequently, we fractioned the silica extract by preparative gas-chromatography (GC) and by separating more polar from less polar compounds. In general, C. marginiventris preferred polar over non-polar compounds, but several fractions were attractive to the wasp, including one that contained compounds emitted in quantities below the detection threshold of the GC analysis. These results imply that the attractiveness of the volatile blend emitted by Spodoptera-infested maize seedlings to C. marginiventris females is determined by a specific combination of attractive and repellent/masking compounds, including some that are emitted in very small amounts. Manipulating the emission of such minor compounds has the potential to greatly improve the attraction of certain parasitoids and enhance biological control of specific insect pests.
The Role of Indole and Other Shikimic Acid Derived Maize Volatiles in the Attraction of Two Parasitic Wasps
After herbivore attack, plants release a plethora of different volatile organic compounds (VOCs), which results in odor blends that are attractive to predators and parasitoids of these herbivores. VOCs in the odor blends emitted by maize plants (Zea mays) infested by lepidopteran larvae are well characterized. They are derived from at least three different biochemical pathways, but the relative importance of each pathway for the production of VOCs that attract parasitic wasps is unknown. Here, we studied the importance of shikimic acid derived VOCs for the attraction of females of the parasitoids Cotesia marginiventris and Microplitis rufiventris. By incubating caterpillar-infested maize plants in glyphosate, an inhibitor of the 5-enolpyruvylshikimate-3-phospate (EPSP) synthase, we obtained induced odor blends with only minute amounts of shikimic acid derived VOCs. In olfactometer bioassays, the inhibited plants were as attractive to naive C. marginiventris females as control plants that released normal amounts of shikimic acid derived VOCs, whereas naive M. rufiventris females preferred inhibited plants to control plants. By adding back synthetic indole, the quantitatively most important shikimic acid derived VOC in induced maize odors, to inhibited plants, we showed that indole had no effect on the attraction of C. marginiventris and that M. rufiventris preferred blends without synthetic indole. Exposing C. marginiventris females either to odor blends of inhibited or control plants during oviposition experiences shifted their preference in subsequent olfactometer tests in favor of the experienced odor. Further learning experiments with synthetic indole showed that C. marginiventris can learn to respond to this compound, but that this does not affect its choices between natural induced blends with or without indole. We hypothesize that for naïve wasps the attractiveness of an herbivore-induced odor blend is reduced due to masking by nonattractive compounds, and that during oviposition experiences in the presence of complex odor blends, parasitoids strongly associate some compounds, whereas others are largely ignored.
Induction of systemic acquired resistance in Zea mays also enhances the plant’s attractiveness to parasitoids
Plants under attack by caterpillars emit volatile compounds that attract the herbivore’s natural enemies. In maize, the caterpillar-induced production of volatiles involves the phytohormone jasmonic acid (JA). In contrast, pathogen attack usually up-regulates the salicylic acid (SA)-pathway and results in systemic acquired resistance (SAR) against plant diseases. Activation of the SA-pathway has often been found to repress JA-dependent direct defenses, but little is known about the effects of SAR induction on indirect defenses such as volatile emission and parasitoid attraction. We examined if induction of SAR in maize, by chemical elicitation with the SA-mimic benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH), attenuates the emission of volatiles induced by Spodoptera littoralis or exogenously applied JA. In addition, we determined how these treatments affected the attractiveness of the plants to the parasitoid Microplitis rufiventris in a six-arm-olfactometer. BTH treatment alone resulted in significant systemic resistance of maize seedlings against the pathogen Setosphaeria turcica, but had no detectable effect on volatile emissions. Induction of SAR significantly reduced the emission rates of two compounds (indole and (E)-β-caryophyllene) in JA-treated plants, whereas no such negative cross-talk was found in caterpillar-damaged plants. Surprisingly, however, BTH treatment prior to caterpillar-feeding made the plants far more attractive to the parasitoid than plants that were only damaged by the herbivore. Control experiments showed that this response was due to plant-mediated effects rather than attractiveness of BTH itself. We conclude that in the studied system, plant protection by SAR activation is compatible with and can even enhance indirect defense against herbivores.