Maria Fernanda G. V. Peñaflor

Weather Forecasting by Insects: Modified Sexual Behaviour in Response to Atmospheric Pressure Changes

Prevailing abiotic conditions may positively or negatively impact insects at both the individual and population levels. For example while moderate rainfall and wind velocity may provide conditions that favour development, as well as movement within and between habitats, high winds and heavy rains can significantly decrease life expectancy. There is some evidence that insects adjust their behaviours associated with flight, mating and foraging in response to changes in barometric pressure. We studied changes in different mating behaviours of three taxonomically unrelated insects, the curcurbit beetle, Diabrotica speciosa (Coleoptera), the true armyworm moth, Pseudaletia unipuncta (Lepidoptera) and the potato aphid, Macrosiphum euphorbiae (Hemiptera), when subjected to natural or experimentally manipulated changes in atmospheric pressure. In response to decreasing barometric pressure, male beetles exhibited decreased locomotory activity in a Y-tube olfactometer with female pheromone extracts. However, when placed in close proximity to females, they exhibited reduced courtship sequences and the precopulatory period. Under the same situations, females of the true armyworm and the potato aphid exhibited significantly reduced calling behaviour. Neither the movement of male beetles nor the calling of armyworm females differed between stable and increasing atmospheric pressure conditions. However, in the case of the armyworm there was a significant decrease in the incidence of mating under rising atmospheric conditions, suggesting an effect on male behaviour. When atmospheric pressure rose, very few M. euphorbiae oviparae called. This was similar to the situation observed under decreasing conditions, and consequently very little mating was observed in this species except under stable conditions. All species exhibited behavioural modifications, but there were interspecific differences related to size-related flight ability and the diel periodicity of mating activity. We postulate that the observed behavioral modifications, especially under decreasing barometric pressure would reduce the probability of injury or death under adverse weather conditions.

A Novel Interaction between Plant-Beneficial Rhizobacteria and Roots: Colonization Induces Corn Resistance against the Root Herbivore Diabrotica speciosa

A number of soil-borne microorganisms, such as mycorrhizal fungi and rhizobacteria, establish mutualistic interactions with plants, which can indirectly affect other organisms. Knowledge of the plant-mediated effects of mutualistic microorganisms is limited to aboveground insects, whereas there is little understanding of what role beneficial soil bacteria may play in plant defense against root herbivory. Here, we establish that colonization by the beneficial rhizobacterium Azospirillum brasilense affects the host selection and performance of the insect Diabrotica speciosa. Root larvae preferentially orient toward the roots of non-inoculated plants versus inoculated roots and gain less weight when feeding on inoculated plants. As inoculation by A. brasilense induces higher emissions of (E)-β-caryophyllene compared with non-inoculated plants, it is plausible that the non-preference of D. speciosa for inoculated plants is related to this sesquiterpene, which is well known to mediate belowground insect-plant interactions. To the best of our knowledge, this is the first study showing that a beneficial rhizobacterium inoculant indirectly alters belowground plant-insect interactions. The role of A. brasilense as part of an integrative pest management (IPM) program for the protection of corn against the South American corn rootworm, D. speciosa, is considered.

Tomato Infection by Whitefly-Transmitted Circulative and Non-Circulative Viruses Induce Contrasting Changes in Plant Volatiles and Vector Behaviour

Virus infection frequently modifies plant phenotypes, leading to changes in behaviour and performance of their insect vectors in a way that transmission is enhanced, although this may not always be the case. Here, we investigated Bemisia tabaci response to tomato plants infected by Tomato chlorosis virus (ToCV), a non-circulative-transmitted crinivirus, and Tomato severe rugose virus (ToSRV), a circulative-transmitted begomovirus. Moreover, we examined the role of visual and olfactory cues in host plant selection by both viruliferous and non-viruliferous B. tabaci. Visual cues alone were assessed as targets for whitefly landing by placing leaves underneath a Plexiglas plate. A dual-choice arena was used to assess whitefly response to virus-infected and mock-inoculated tomato leaves under light and dark conditions. Thereafter, we tested the whitefly response to volatiles using an active air-flow Y-tube olfactometer, and chemically characterized the blends using gas chromatography coupled to mass spectrometry. Visual stimuli tests showed that whiteflies, irrespective of their infectious status, always preferred to land on virus-infected rather than on mock-inoculated leaves. Furthermore, whiteflies had no preference for either virus-infected or mock-inoculated leaves under dark conditions, but preferred virus-infected leaves in the presence of light. ToSRV-infection promoted a sharp decline in the concentration of some tomato volatiles, while an increase in the emission of some terpenes after ToCV infection was found. ToSRV-viruliferous whiteflies preferred volatiles emitted from mock-inoculated plants, a conducive behaviour to enhance virus spread, while volatiles from ToCV-infected plants were avoided by non-viruliferous whiteflies, a behaviour that is likely detrimental to the secondary spread of the virus. In conclusion, the circulative persistent begomovirus, ToSRV, seems to have evolved together with its vector B. tabaci to optimise its own spread. However, this type of virus-induced manipulation of vector behaviour was not observed for the semi persistent crinivirus, ToCV, which is not specifically transmitted by B. tabaci and has a much less intimate virus-vector relationship.

Role of Methyl Salicylate on Oviposition Deterrence in Arabidopsis thaliana

Plants attacked by herbivores have evolved different strategies that fend off their enemies. Insect eggs deposited on leaves have been shown to inhibit further oviposition through visual or chemical cues. In some plant species, the volatile methyl salicylate (MeSA) repels gravid insects but whether it plays the same role in the model species Arabidopsis thaliana is currently unknown. Here we showed that Pieris brassicae butterflies laid fewer eggs on Arabidopsis plants that were next to a MeSA dispenser or on plants with constitutively high MeSA emission than on control plants. Surprisingly, the MeSA biosynthesis mutant bsmt1-1 treated with egg extract was still repellent to butterflies when compared to untreated bsmt1-1. Moreover, the expression of BSMT1 was not enhanced by egg extract treatment but was induced by herbivory. Altogether, these results provide evidence that the deterring activity of eggs on gravid butterflies is independent of MeSA emission in Arabidopsis, and that MeSA might rather serve as a deterrent in plants challenged by feeding larvae.

Effects of single and multiple herbivory by host and non-host caterpillars on the attractiveness of herbivore-induced volatiles of sugarcane to the generalist parasitoid Cotesia flavipes

It is well known that parasitoids are attracted to volatiles emitted by host‐damaged plants; however, this tritrophic interaction may change if plants are attacked by more than one herbivore species. The larval parasitoid Cotesia flavipesCameron (Hymenoptera: Braconidae) has been used intensively in Brazil to control the sugarcane borer, Diatraea saccharalisFabricius (Lepidoptera: Pyralidae) in sugarcane crops, where Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), a non‐stemborer lepidopteran, is also a pest. Here, we investigated the ability of C. flavipes to discriminate between an unsuitable host (S. frugiperda) and a suitable host (D. saccharalis) based on herbivore‐induced plant volatiles (HIPVs) emitted by sugarcane, and whether multiple herbivory (D. saccharalis feeding on stalk + S. frugiperda feeding on leaves) in sugarcane affected the attractiveness of HIPVs to C. flavipes. Olfactometer assays indicated that volatiles of host and non‐host‐damaged plants were attractive to C. flavipes. Even though host‐ and non‐host‐damaged plants emitted considerably different volatile blends, neither naïve nor experienced wasps discriminated suitable and unsuitable hosts by means of HIPVs emitted by sugarcane. With regard to multiple herbivory, wasps innately preferred the odor blend emitted by sugarcane upon non‐host + host herbivory over host‐only damaged plants. Multiple herbivory caused a suppression of some volatiles relative to non‐host‐damaged sugarcane that may have resulted from the unaltered levels of jasmonic acid in host‐damaged plants, or from reduced palatability of host‐damaged plants to S. frugiperda. In conclusion, our study showed that C. flavipes responds to a wide range of plant volatile blends, and does not discriminate host from non‐host and non‐stemborer caterpillars based on HIPVs emitted from sugarcane. Moreover, we showed that multiple herbivory by the sugarcane borer and fall armyworm increases the attractiveness of sugarcane plants to the parasitoids.

Attraction of entomopathogenic nematodes to sugarcane root volatiles under herbivory by a sap-sucking insect

Few systems have been described in which herbivore-induced root volatiles mediate attraction of entomopathogenic nematodes (EPNs), and they only concern root damage inflicted by chewing insects. EPNs, especially Heterorhabditis indica and Steinernema carpocapsae, are potential biological control agents of sugarcane spittlebug (Mahanarva fimbriolata) populations. Here, we investigated the response of these two species of EPNs to sugarcane root volatiles damaged by M. fimbriolata nymphs in a belowground six-arm olfactometer. We also examined changes on root volatile profile in response to herbivory of sugarcane spittlebug nymphs. Results showed that both EPN species did not discriminate between odors of undamaged sugarcane and moistened sand (blank). However, when EPNs were exposed to odors of spittlebug-damaged and undamaged sugarcane roots, both species significantly preferred odors of spittlebug-damaged roots. Headspace collection followed by GC–MS analyses showed no qualitative difference (total of 11 compounds) between volatile profiles of spittlebug-damaged and undamaged sugarcane roots. In contrast to the previous studies involving feeding by root chewing insects, our root volatile analysis did not reveal any up-regulation resulting from sugarcane spittlebug damage, but the down-regulation of the terpenes dihydromyrcenol and β-isomethyl ionone when compared with the profile of undamaged sugarcane roots. Here, we propose alternative explanations for the EPN attraction to spittlebug-damaged roots as it is unlikely that reduced concentrations of the volatiles play a role in this interaction. Further studies are necessary to determine the key compounds of the root volatile emission to enhance biological control efficacy with EPNs against M. fimbriolata in sugarcane.

The impact of coffee and pasture agriculture on predatory and omnivorous leaf-litter ants

Abstract Ants are known to function as reliable biological indicators for habitat impact assessment. They play a wide range of ecological roles depending on their feeding and nesting habits. By clustering ants in guilds, it is possible both to assess how agriculture and forest fragmentation can disturb ant communities and to predict the ecological impacts due to losses of a specific guild. This study aimed at determining the impact of non-shaded coffee and pasture agriculture on predatory and omnivorous guilds of leaf-litter ants of Atlantic Forest fragments in Minas Gerais, Brazil. Both coffee and pasture agriculture influenced leaf-litter ant community, although coffee was more disruptive than pasture. Coffee agriculture not only disturbed the diversity of predatory ants, but also negatively affected the number of predatory and omnivorous ants when compared to forest fragments. In contrast, pasture agriculture only disrupted the abundance of predatory ants. Fragment edges skirting crops were negatively affected in terms of leaf-litter ant abundance, but not diversity. Cluster analysis showed that forest fragments were similar irrespective of the cultivation, but the borders were similar to the crop. The study assessed agriculture impact by surveying ant guilds, and revealed that the predatory guild is more susceptible than omnivorous ants.

Nocturnal herbivore-induced plant volatiles attract the generalist predatory earwig Doru luteipes Scudder

Numerous studies have demonstrated that entomophagous arthropods use herbivore-induced plant volatile (HIPV) blends to search for their prey or host. However, no study has yet focused on the response of nocturnal predators to volatile blends emitted by prey damaged plants. We investigated the olfactory behavioral responses of the night-active generalist predatory earwig Doru luteipes Scudder (Dermaptera: Forficulidae) to diurnal and nocturnal volatile blends emitted by maize plants (Zea mays) attacked by either a stem borer (Diatraea saccharalis) or a leaf-chewing caterpillar (Spodoptera frugiperda), both suitable lepidopteran prey. Additionally, we examined whether the earwig preferred odors emitted from short- or long-term damaged maize. We first determined the earwig diel foraging rhythm and confirmed that D. luteipes is a nocturnal predator. Olfactometer assays showed that during the day, although the earwigs were walking actively, they did not discriminate the volatiles of undamaged maize plants from those of herbivore damaged maize plants. In contrast, at night, earwigs preferred volatiles emitted by maize plants attacked by D. saccharalis or S. frugiperda over undamaged plants and short- over long-term damaged maize. Our GC-MS analysis revealed that short-term damaged nocturnal plant volatile blends were comprised mainly of fatty acid derivatives (i.e., green leaf volatiles), while the long-term damaged plant volatile blend contained mostly terpenoids. We also observed distinct volatile blend composition emitted by maize damaged by the different caterpillars. Our results showed that D. luteipes innately uses nocturnal herbivore-induced plant volatiles to search for prey. Moreover, the attraction of the earwig to short-term damaged plants is likely mediated by fatty acid derivatives.

Direct and indirect resistance of sugarcane to Diatraea saccharalis induced by jasmonic acid

Treating plants with synthetic jasmonic acid (JA) induces a defensive response similar to herbivore attack, and is a potential strategy for integrated pest management. Despite the importance of sugarcane, its JA-induced defences have not yet been studied. We investigated the effects of JA treatment on the direct and indirect resistance of sugarcane to the key-pest and specialist herbivore Diatraea saccharalis and the generalist Spodoptera frugiperda. Indirect defences were examined by testing the attraction of Cotesia flavipes, a sugarcane-borer parasitoid, to JA-induced volatile. The results showed that JA-treated sugarcane did not affect the weight gain of the two larvae. However, in dual-choice assays, both species preferred to feed on mock rather than JA-treated plants. Leaf colorimetric analyses showed that visual cues are unlikely to be involved in larval preference, whereas results from olfactometric assays revealed that D. saccharalis preferred JA-induced over mock plant volatiles. After 48 h of treatment, JA-treated plants emitted a volatile blend attractive to C. flavipes, comprised mainly of sesquiterpenes. However, the parasitoid did not discriminate JA-treated from host-damaged plant volatiles. When the wasps were given a choice between JA-treated and JA-treated + host-damaged plants, they preferred the latter, which emitted a more complex blend, suggesting that JA treatment likely does not hamper host-finding. We concluded that JA induces the emission of volatiles that are attractive to the sugarcane borer parasitoid, as well as an antixenosis type of resistance in sugarcane against the two pests, although neither volatiles nor visual cues alone are involved in the underlying mechanism.

The effects of Gibberella zeae, Barley Yellow Dwarf Virus, and co-infection on Rhopalosiphum padi olfactory preference and performance

Insect-borne viruses promote several changes in plant phenotype, which can modify plant-vector interactions in favor of virus survival and dissemination. Although co-infections commonly occur in the field, little is known about their effects on interactions with the vector. The ecological interactions between Barley Yellow Dwarf Virus (BYDV) and its aphid vector, Rhopalosiphum padi, have been investigated extensively, but the vector’s behavior in more complex scenarios has yet to be examined. We assessed olfactory response and performance of R. padi to wheat singly and doubly infected by the pathogenic fungus Giberella zeae and BYDV. Non-viruliferous aphids preferred odors of BYDV-infected wheat over healthy wheat, as previously reported in the literature, and they were still preferentially attracted to BYDV-infected plant during co-infection. However, around 35% more non-viruliferous aphids chose healthy wheat over G. zeae-infected wheat. Viruliferous aphids did not show any preference to the treatments. BYDV-infected wheat was a superior host than healthy wheat for the aphids whose population increased in 25%. We observed a synergistic effect of the co-infected wheat, which was the best host for aphids, and promoted an elevation of 42% on population growth. Our results indicate that co-infection might be beneficial for virus spread as does not interfere with aphid olfactory preference and provides greater colony growth than in singly infected plants.