Field evidence for the role of plant volatiles induced by caterpillar oral secretion in prey localization by predatory social wasps.

One assumed function of herbivore-induced plant volatiles (HIPVs) is to attract natural enemies of the inducing herbivores. Field evidence for this is scarce. In addition, the assumption that elicitors in oral secretions that trigger the volatile emissions are essential for the attraction of natural enemies has not yet been demonstrated under field conditions. After observing predatory social wasps removing caterpillars from maize plants, we hypothesized that these wasps use HIPVs to locate their prey. To test this, we conducted an experiment that simultaneously explored the importance of caterpillar oral secretions in the interaction. Spodoptera caterpillars pinned onto mechanically damaged plants treated with oral secretion were more likely to be attacked by wasps compared with caterpillars on plants that were only mechanically wounded. Both of the latter treatments were considerably more attractive than plants only treated with oral secretion or left untreated. Subsequent analyses of headspace volatiles confirmed differences in emitted volatiles that likely account for the differential predation across treatments. These findings highlight the importance of HIPVs in prey localization by social wasps, hitherto underappreciated potential biocontrol agents and provide evidence for the role that elicitors play in inducing attractive odour blends.

Entomopathogenic Nematodes-Killed Insect Cadavers in the Rhizosphere Activate Plant Direct and Indirect Defences Aboveground.

Plants can perceive and respond to external stimuli by activating both direct and indirect defences against herbivores. Soil-dwelling entomopathogenic nematodes (EPNs), natural enemies of root-feeding herbivores, carry symbiotic bacteria that grow and reproduce once inside arthropod hosts. We hypothesized that the metabolites produced by EPN-infected insect cadavers could be perceived by plants, thereby activating plant defences systemically. We tested this hypothesis by adding three EPN-infected Galleria mellonella cadavers to maize plants and testing plant responses against a major maize pest (Spodoptera frugiperda) and one of its parasitoids (Trichogramma dendrolimi). We found that S. frugiperda females deposited fewer, and caterpillars fed less on maize plants growing near EPN-infected cadavers than on control plants. Accordingly, EPN-infected cadavers triggered the systemic accumulation of defence hormones (SA), genes (PR1), and enzymes (SOD, POD, and CAT) in maize leaves. Furthermore, four volatile organic compounds produced by plants exposed to EPN-infected cadavers deterred S. frugiperda caterpillars and female adults. However, these compounds were more attractive to T. dendrolimi parasitoids. Our study enhances the understanding of the intricate relationships within the above- and belowground ecosystems and provides crucial insights for advancing sustainable pest management strategies.

Dose-dependent dynamics of densovirus infection in two nymphalid butterfly species utilizing native or exotic host plants.

Insects are attacked by a diverse range of microbial pathogens in the wild. In herbivorous species, larval host plants frequently play a critical role in mediating susceptibility to infection. Characterizing such plant-mediated effects on herbivore-pathogen interactions can provide insight into patterns of infection across wild populations. In this study, we investigated the effects of host plant use by two North American butterflies, Euphydryas phaeton (Nymphalidae) and Anartia jatrophae (Nymphalidae), on entomopathogen infection across a range of three doses. Both of these herbivores recently incorporated the same exotic plant, Plantago lanceolata (Plantaginaceae), into their host range and are naturally infected by the same entomopathogen, Junonia coenia densovirus (Parvoviridae), in wild populations. We performed two factorial experiments in which E. phaeton and A. jatrophae were reared on either P. lanceolata or a native host plant [Chelone glabra (Plantaginaceae) for E. phaeton; Bacopa monnieri (Plantaginaceae) for A. jatrophae] and inoculated with either a low, medium, or high dose of the virus. In E. phaeton, the outcomes of infection were highly dose-dependent, with inoculation with higher viral doses resulting in faster time to death and greater mortality. However, neither survival nor postmortem viral burdens varied depending upon the host plant that was consumed. In contrast, host plant use had a strong effect on viral burdens in A. jatrophae, with consumption of the exotic plant appearing to enhance host resistance to infection. Together, these results illustrate the variable influences of host plant use on herbivore resistance to infection, highlighting the importance of investigating plant-herbivore relationships within a tritrophic framework.

Plant resistance and predators influence the density dependence of herbivore survival and distribution of herbivory.

Plant resistance and predators can influence density-dependent survivorship and growth of herbivores, and their damage to plants. Although the independent effects of plant resistance and predators on herbivores and herbivory are well known, little is known about their interactive and density-dependent effects on herbivores and the amount and distribution of damage on plants. These relationships are important for understanding how herbivore and plant populations influence each other. We used a laboratory density-manipulation experiment to determine how plant resistance (three treatments: jasmonate-insensitive, unmanipulated wild type, and jasmonate-sprayed wild-type plants) and predation (two treatments: predator or no predator) affect the survivorship and growth of an herbivore, as well as per capita damage and the distribution of damage on plants. We found evidence that the density dependence of herbivore survivorship was influenced by predators and an interactive effect of plant resistance and predation. Herbivore growth was reduced by higher plant resistance but was not density-dependent nor affected by predation. Per capita plant damage was reduced by plant resistance, predation, and herbivore density. The within-plant distribution of damage became more even with increasing herbivore density but was not affected by predation or the independent effect of plant resistance. The distribution of damage was also affected by an interaction between plant resistance and herbivore density; damage became less aggregated with density across all plant resistance treatments, but the decrease was strongest for the jasmonate-insensitive plants. These results show that predators influence herbivore density dependence, and that plant resistance can affect the impact of predators on herbivores. Though plant resistance, predation, and herbivore density all reduced per capita herbivore damage to plants, only herbivore density and plant resistance affected the distribution of damage. Distributions of herbivory can influence plant success; documenting patterns of herbivory is an under-appreciated avenue for integrating effects of plant resistance, predators, and herbivore density on plant-herbivore interactions.

Small but strong: herbivory by sap-feeding insect reduces plant progeny growth but enhances direct and indirect anti-herbivore defenses.

The transmission of resistance traits to herbivores across subsequent generations is an important strategy employed by plants to enhance their fitness in environments with high herbivore pressure. However, our understanding of the impact of maternal herbivory on direct and indirect induced chemical defenses of progeny, as well as the associated costs, is currently limited to herbivory by leaf-chewing insects. In this study, we investigated the transgenerational effects of a sap-feeding insect, the green peach aphid Myzus persicae, on direct and indirect chemical defenses of bell pepper plants (Capsicum annuum), and whether the effects entail costs to plant growth. Aphid herbivory on parental plants led to a reduced number of seeds per fruit, which exhibited lower germination rates and produced smaller seedlings compared to those from non-infested parental plants. In contrast, the progeny of aphid-infested plants were less preferred as hosts by aphids and less suitable than the progeny of non-infested plants. This enhanced resistance in the progeny of aphid-infested plants coincided with elevated levels of both constitutive and herbivore-induced total phenolic compounds, compared to the progeny of non-infested plants. Furthermore, the progeny of aphid-infested plants emitted herbivore-induced plant volatiles (HIPVs) that were more attractive to the aphid parasitoid Aphidius platensis than those emitted by the progeny of non-infested plants. Our results indicate that herbivory by sap-feeding insect induces transgenerational resistance on progeny bell pepper plants, albeit at the expense of vegetative growth.

Host plant-mediation of viral transmission and its consequences for a native butterfly.

Pathogens play a key role in insect population dynamics, contributing to short-term fluctuations in abundance as well as long-term demographic trends. Two key factors that influence the effects of entomopathogens on herbivorous insect populations are modes of pathogen transmission and larval host plants. In this study, we examined tritrophic interactions between a sequestering specialist lepidopteran, Euphydryas phaeton, and a viral pathogen, Junonia coenia densovirus, on its native host plant, Chelone glabra, and a novel host plant, Plantago lanceolata, to explore whether host plant mediates viral transmission, survival, and viral loads. A two-factor factorial experiment was conducted in the laboratory with natal larval clusters randomly assigned to either the native or novel host plant and crossed with either uninoculated controls or viral inoculation (20% of individuals in the cluster inoculated). Diapausing clusters were overwintered in the laboratory and checked weekly for mortality. At the end of diapause, all surviving individuals were reared to adulthood to estimate survivorship. All individuals were screened to quantify viral loads, and estimate horizontal transmission postmortem. To test for vertical transmission, adults were mated, and the progeny were screened for viral presence. Within virus-treated groups, we found evidence for both horizontal and vertical transmission. Larval clusters reared on the native host plant had slightly higher horizontal transmission. Survival probability was lower in clusters feeding on the native host plant, with inoculated groups reared on the native host plant experiencing complete mortality. Viral loads did not differ by the host plant, although viral loads decreased with increased sequestration of secondary compounds on both host plants. Our results indicate that the use of a novel host plant may confer fitness benefits in terms of survival and reduced viral transmission when larvae feeding on it are infected with this pathogen, supporting hypotheses of potential evolutionary advantages of a host range expansion in the context of tritrophic interactions.

Urban socioeconomic variation influences the ecology and evolution of trophic interactions.

As urbanization expands, it is becoming increasingly important to understand how anthropogenic activity is affecting ecological and evolutionary processes. Few studies have examined how human social patterns within cities can modify eco-evolutionary dynamics. We tested how socioeconomic variation corresponds with changes in trophic interactions and natural selection on prey phenotypes using the classic interaction between goldenrod gall flies (Eurosta solidaginis) and their natural enemies: birds, beetles, and parasitoid wasps. We sampled galls from 84 sites across neighbourhoods with varying socioeconomic levels, and quantified the frequency of predation/parasitism on flies and natural selection by each enemy. We found that bird predation was higher in the highest income neighbourhoods, increasing the strength of selection for smaller galls. Wasp and beetle attack, but not their strength of selection, increased in lower income neighbourhoods. We show that socioeconomic variation in cities can have strong unintended consequences for the ecology and evolution of trophic interactions.

Simulated Herbivory Affects the Volatile Emissions of Oak Saplings, while Neighbourhood Affects Flavan-3-ols Content of Their Leaves.

To what extent particular plant defences against herbivorous insects are constitutive or inducible will depend on the costs and benefits in their neighbourhood. Some defensive chemicals in leaves are thought to be costly and hard to produce rapidly, while others, including volatile organic compounds that attract natural enemies, might be cheaper and can be released rapidly. When surrounding tree species are more closely related, trees can face an increased abundance of both specialist herbivores and their parasitoids, potentially increasing the benefits of constitutive and inducible defences. To test if oaks (Quercus robur) respond more to herbivore attacks with volatile emission than with changes in leaf phenolic chemistry and carbon to nitrogen ratio (C: N), and whether oaks respond to the neighbouring tree species, we performed an experiment in a forest in Poland. Oak saplings were placed in neighbourhoods dominated by oak, beech, or pine trees, and half of them were treated with the phytohormone methyl jasmonate (elicitor of anti-herbivore responses). Oaks responded to the treatment by emitting a different volatile blend within 24 h, while leaf phenolic chemistry and C: N remained largely unaffected after 16 days and multiple treatments. Leaf phenolics were subtly affected by the neighbouring trees with elevated flavan-3-ols concentrations in pine-dominated plots. Our results suggest that these oaks rely on phenols as a constitutive defence and when attacked emit volatiles to attract natural enemies. Further studies might determine if the small effect of the neighbourhood on leaf phenolics is a response to different levels of shading, or if oaks use volatile cues to assess the composition of their neighbourhood.

Natural habitat connectivity and organic management modulate pest dispersal, gene flow, and natural enemy communities.

The simplification and fragmentation of agricultural landscapes generate effects on insects at multiple spatial scales. As each functional group perceives and uses the habitat differently, the response of pest insects and their associated natural enemies to environmental changes varies. Therefore, landscape structure may have consequences on gene flow among pest populations in space. This study aimed to evaluate the effects of local and landscape factors, at multiple scales, on the local infestation, gene flow and broad dispersion dynamics of the pest insect Bemisia tabaci (Genn.) Middle East-Asia Minor 1 (MEAM-1, former biotype B) (Hemiptera: Aleyrodidae) and its associated natural enemies in a tropical agroecosystem. We evaluated the abundance of B. tabaci populations and their natural enemy community in 20 tomato farms in Brazil and the gene flow between farms from 2019 to 2021. Landscapes dominated by agriculture resulted in larger B. tabaci populations and higher gene flow, especially in conventional farms. A higher density of native vegetation patches disfavored pest populations, regardless of the management system. The results revealed that whitefly responds to intermediate spatial scales and that landscape factors interact with management systems to modulate whitefly populations on focal farms. Conversely, whitefly natural enemies benefited from higher amounts of natural vegetation at small spatial scales, while the connectivity between natural habitat patches was beneficial for natural enemies regardless of the distance from the focal farm. The resulting dispersion model predicts that the movement of whiteflies between farms increases as the amount of natural vegetation decreases. Our findings demonstrate that landscape features, notably landscape configuration, can mediate infestation episodes, as they affect pest insects and natural enemies in opposite ways. We also showed that landscape features interact with farm traits, which highlights the need for management strategies at multiple spatial scales. In conclusion, we demonstrated the importance of the conservation of natural areas as a key strategy for area-wide ecological pest management and the relevance of organic farming to benefit natural enemy communities in tropical agroecosystems.

Unveiling the Slippery Secrets of Saliva: Effector Proteins of Phloem-Feeding Insects.

Phloem-feeding insects include many important agricultural pests that cause crop damage globally, either through feeding-related damage or upon transmission of viruses and microbes that cause plant diseases. With genetic crop resistances being limited to most of these pests, control relies on insecticides, which are costly and damaging to the environment and to which insects can develop resistance. Like other plant parasites, phloem-feeding insects deliver effectors inside their host plants to promote susceptibility, most likely by a combination of suppressing immunity and promoting nutrient availability. The recent emergence of the effector paradigm in plant-insect interactions is highlighted by increasing availability of effector repertoires for a range of species and a broadening of our knowledge concerning effector functions. Here, we focus on recent progress made toward identification of effector repertoires from phloem-feeding insects and developments in effector biology that will advance functional characterization studies. Importantly, identification of effector activities from herbivorous insects promises to provide new avenues toward development of crop protection strategies. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Identity Matters: Multiple Herbivory Induces Less Attractive or Repellent Coffee Plant Volatile Emission to Different Natural Enemies.

Co-infestations by herbivores, a common situation found in natural settings, can distinctly affect induced plant defenses compared to single infestations. Related tritrophic interactions might be affected through the emission of changed blends of herbivore-induced plant volatiles (HIPVs). In a previous study, we observed that the infestation by red spider mite (Oligonychus ilicis) on coffee plants facilitated the infestation by white mealybug (Planococcus minor), whereas the reverse sequence of infestation did not occur. Here, we examined the involvement of the jasmonate and salicylate pathways in the plant-mediated asymmetrical facilitation between red spider mites and white mealybugs as well as the effect of multiple herbivory on attractiveness to the predatory mite Euseius concordis and the ladybug Cryptolaemus montrouzieri. Both mite and mealybug herbivory led to the accumulation of JA-Ile, JA, and cis-OPDA in plants, although the catabolic reactions of JA-Ile were specifically regulated by each herbivore. Infestation by mites or mealybugs induced the release of novel volatiles by coffee plants, which selectively attracted their respective predators. Even though the co-infestation by mites and mealybugs resulted in a stronger accumulation of JA-Ile, JA and SA than the single infestation treatments, the volatile emission was similar to that of mite-infested or mealybug-infested plants. However, multiple infestation had a negative impact on the attractiveness of HIPVs to the predators, making them less attractive to the predatory mite and a repellent to the ladybug. We discuss the potential underlying mechanisms of the susceptibility induced by mites, and the effect of multiple infestation on each predator.

Indirect plant-mediated interactions between heterospecific parasitoids that develop in different caterpillar species.

Parasitoids induce physiological changes in their herbivorous hosts that affect how plants respond to herbivory. The signature of parasitoids on induced plant responses to feeding by parasitized herbivores indirectly impacts insect communities interacting with the plant. The effect may extend to parasitoids and cause indirect interaction between parasitoids that develop inside different herbivore hosts sharing the food plant. However, this type of interactions among parasitoid larvae has received very little attention. In this study, we investigated sequential and simultaneous plant-mediated interactions among two host-parasitoid systems feeding on Brassica oleracea plants: Mamestra brassicae parasitized by Microplitis mediator and Pieris rapae parasitized by Cotesia rubecula. We measured the mortality, development time, and weight of unparasitized herbivores and performance of parasitoids that had developed inside the two herbivore species when sharing the food plant either simultaneously or sequentially. Plant induction by parasitized or unparasitized hosts had no significant effect on the performance of the two herbivore host species. In contrast, the two parasitoid species had asymmetrical indirect plant-mediated effects on each other's performance. Cotesia rubecula weight was 15% higher on plants induced by M. mediator-parasitized hosts, compared to control plants. In addition, M. mediator development time was reduced by 30% on plants induced by conspecific but not heterospecific parasitoids, compared to plants induced by its unparasitized host. Contrary to sequential feeding, parasitoids had no effect on each other's performance when feeding simultaneously. These results reveal that indirect plant-mediated interactions among parasitoid larvae could involve any parasitoid species whose hosts share a food plant.

Flavonoid production in tomato mediates both direct and indirect plant defences against whiteflies in tritrophic interactions.

BACKGROUND: The role of plant flavonoids in direct defences against chewing and sap-sucking herbivorous insects has been extensively characterized. However, little is known about flavonoid-mediated tritrophic interactions between plants, herbivorous insects and natural enemies. In this study, we investigated how flavonoids modulate plant-insect interactions in a tritrophic system involving near-isogenic lines (NILs) of cultivated tomato (Solanum lycopersicum) with high (line NIL-purple hypocotyl [PH]) and low (line NIL-green hypocotyl [GH]) flavonoid levels, with a generalist herbivore whitefly (Bemisia tabaci) and its predatory bug (Orius sauteri). RESULTS: By contrasting levels of tomato flavonoids (direct defence) while manipulating the presence of predators (indirect defence), we found that high production of flavonoids in tomato was associated with a higher inducibility of direct defences and a stronger plant resistance to whitefly infestation and stimulated the emissions of induced volatile organic compounds, thereby increasing the attractiveness of B. tabaci-infested plants to the predator O. sauteri. Furthermore, suppression of B. tabaci population growth and enhancement of plant growth were mediated directly by the high production of flavonoids and indirectly by the attraction of O. sauteri, and the combined effects were larger than each effect individually. CONCLUSION: Our results show that high flavonoid production in tomato enhances herbivore-induced direct and indirect defences to better defend against herbivores in tritrophic interactions. Thus, the development of transgenic plants may present an opportunity to utilize the beneficial role of flavonoids in integrated pest management, while simultaneously maintaining or improving resistance against other pests and pathogens. © 2023 Society of Chemical Industry.

Fruit Flies (Diptera: Tephritoidea) and Parasitoids (Hymenoptera) Associated with Native Fruit Trees in the Chaco Biome.

Fruit flies (Diptera: Tephritoidea) are the main pests over important fruits and vegetables. In this research was evaluated the tritrophic interactions of fruit flies and their parasitoids occurring in native fruits in the Chaco Biome. Fruit sampling was carried out monthly in three areas of vegetation communities in Chaco Biome: Forested Steppic Savanna, Wooded Steppic Savanna, Park Steppic Savanna, Porto Murtinho-MS, Brazil, from April 3, 2017, to November 16, 2018 (totaling 20 samples). Fruits of 33 plant species were examined for fruit flies and parasitoids from these three Chaco locations. Sixteen species of fruit plants were infested by 11 species of fruit flies: five species of Anastrepha Schiner (Tephritidae): Anastrepha fraterculus (Wiedemann), Anastrepha obliqua (Macquart), Anastrepha sororcula Zucchi, Anastrepha turpiniae Stone, Anastrepha zenildae Zucchi, and six of Neosilba McAlpine (Lonchaeidae): Neosilba bifida Strikis and Prado, Neosilba certa (Walker), Neosilba glaberrima (Wiedemann), Neosilba inesperata Strikis and Prado, Neosilba pendula (Bezzi), Neosilba zadolicha McAlpine and Steyskal. Three species of parasitoids: Doryctobracon areolatus (Szepliget) and Utetes anastrephae (Viereck) (Braconidae) parasitized Anastrepha spp., and Aganaspis pelleranoi (Figitidae) parasitized Neosilba spp. All fruit flies and parasitoid species reported here are new records for the Chaco Biome. Furthermore, the following trophic associations are new records worldwide: Anastrepha obliqua in Sideroxylon obtusifolium; Anastrepha zenildae, Neosilba inesperata, and Neosilba zadolicha in Eugenia myrcianthes; Anastrepha fraterculus, Anastrepha sororcula, Neosilba pendula, and Neosilba inesperata in Campomanesia adamantium; Anastrepha spp. in Garcinia gardneriana and Agonandra brasiliensis.

Sequestration of cucurbitacins from cucumber plants by Diabrotica balteata larvae provides little protection against biological control agents.

Cucurbitaceae plants produce cucurbitacins, bitter triterpenoids, to protect themselves against various insects and pathogens. Adult banded cucumber beetles (Diabrotica balteata), a common pest of maize and cucurbits, sequester cucurbitacins, presumably as a defensive mechanism against their natural enemies, which might reduce the efficacy of biological control agents. Whether the larvae also sequester and are protected by cucurbitacins is unclear. We profiled cucurbitacin levels in four varieties of cucumber, Cucumis sativus, and in larvae fed on these varieties. Then, we evaluated larval growth and resistance against common biocontrol organisms including insect predators, entomopathogenic nematodes, fungi and bacteria. We found considerable qualitative and quantitative differences in the cucurbitacin levels of the four cucumber varieties. While two varieties were fully impaired in their production, the other two accumulated high levels of cucurbitacins. We also observed that D. balteata larvae sequester and metabolize cucurbitacins, and although the larvae fed extensively on both belowground and aboveground tissues, the sequestered cucurbitacins were mainly derived from belowground tissues. Cucurbitacins had no detrimental effects on larval performance and, surprisingly, did not provide protection against any of the natural enemies evaluated. Our results show that D. balteata larvae can indeed sequester and transform cucurbitacins, but sequestered cucurbitacins do not impact the biocontrol potential of common natural enemies used in biocontrol. Hence, this plant trait should be conserved in plant breeding programs, as it has been demonstrated in previous studies that it can provide protection against plant pathogens and generalist insects. Supplementary Information: The online version contains supplementary material available at 10.1007/s10340-022-01568-3.

Use of an exotic host plant reduces viral burden in a native insect herbivore.

Incorporation of exotic plants into the diets of native herbivores is a common phenomenon, influencing interactions with natural enemies and providing insight into the tritrophic costs and benefits of dietary expansion. We evaluated how use of an exotic plant, Plantago lanceolata, impacted immune performance, development and susceptibility to pathogen infection in the neotropical herbivore Anartia jatrophae (Lepidoptera: Nymphalidae). Caterpillars were reared on P. lanceolata or a native plant, Bacopa monnieri, and experimentally infected with a pathogenic virus, Junonia coenia densovirus. We found that virus-challenged herbivores exhibited higher survival rates and lower viral burdens when reared on P. lanceolata compared to B. monnieri, though immune performance and development time were largely similar on the two plants. These findings reveal that use of an exotic plant can impact the vulnerability of a native herbivore to pathogen infection, suggesting diet-mediated protection against disease as a potential mechanism facilitating the incorporation of novel resources.

Prey-mediated effects of mCry51Aa2-producing cotton on the predatory nontarget bug Orius majusculus (Reuter).

Genetically engineered (GE) cotton, MON 88702, is protected against certain sucking pests, such as plant bugs and thrips, by producing mCry51Aa2, a modified protein from Bacillus thuringiensis (Bt). Predatory pirate bugs (Orius spp.), natural enemies contributing to biological pest control, are also sensitive to the insecticidal protein when exposed continuously to high concentrations. We evaluated effects of MON 88702 on Orius majusculus when fed prey types with different mCry51Aa2 concentrations. When neonates were provided exclusively Tetranychus urticae spider mites reared on MON 88702 (high mCry51Aa2 content), adverse effects on predator survival and development were confirmed, compared with specimens fed prey from near-isogenic non-Bt cotton. When fed a mixture of T. urticae and Ephestia kuehniella eggs (mCry51Aa2-free), predator life table parameters were similar to the treatment where eggs were fed exclusively. When mCry51Aa2-containing spider mites were provided for a limited time at the beginning or the end of juvenile development, effects were less pronounced. While pirate bug nymphs showed similar consumption rates for prey from Bt and non-Bt cotton, choice experiments revealed a preference for E. kuehniella eggs over spider mites. Lepidopteran larvae (Spodoptera littoralis, high mCry51Aa2 content) or cotton aphids (Aphis gossypii, mCry51Aa2-free) reared on MON 88702 as alternative prey did not result in adverse effects on O. majusculus. Our study suggests limited risk of mCry51Aa2-producing cotton for O. majusculus, because its sensitivity for the Bt protein is relatively low and its natural food consists of diverse prey species with varying concentrations of Bt protein.

1-nonene plays an important role in the response of maize-aphid-ladybird tritrophic interactions to nitrogen.

Plant volatile organic compounds (VOCs) are the key distress signals involved in tritrophic interactions, by which plants recruit predators to protect themselves from herbivores. However, the effect of nitrogen fertilization on VOCs that mediate tritrophic interactions remains largely unidentified. In this study, a maize (Zea mays)-aphid (Rhopalosiphum padi)-ladybird (Harmonia axyridis) tritrophic interaction model was constructed under high-nitrogen (HN) and low-nitrogen (LN) regimens. H. axyridis had a stronger tendency to be attracted by aphid-infested maize under HN conditions. Then, volatiles were collected and identified from maize leaves on which aphids had fed. All of the HN-induced volatiles (HNIVs) elicited an electroantennogram (EAG) response from H. axyridis. Of these HNIVs, 1-nonene was attractive to H. axyridis under simulated natural volatilization. Furthermore, our regression showed that the release of 1-nonene was positively correlated with H. axyridis visitation rates. Supplying 1-nonene to maize on which aphids had fed under LN enhanced attractiveness to H. axyridis. These results supported the conclusion that 1-nonene was the active compound that mediated the response to nitrogen in the tritrophic interaction. In addition, the 1-nonene synthesis pathway was hypothesized, and we found that the release of 1-nonene might be related to the presence of salicylic acid (SA) and abscisic acid (ABA). This research contributes to the development of novel environmentally friendly strategies to optimize nitrogen fertilizer application and to improve pest control in maize crops.

Safety of Bacillus thuringiensis Cry1Ah and Vip3Aa toxins for the predatory stink bug Arma custos (Hemiptera: Pentatomidae).

The widespread adoption of Bt crops expressing insecticidal proteins derived from Bacillus thuringiensis has created a need to assess the potential effects of these toxins on non-target organisms, especially species such as Arma custos, a generalist predator that provides important biological control services in many field crops in Asia. Direct dietary exposure of A. custos to Cry1Ah and Vip3Aa proteins produced no adverse effects on life history traits, despite continuous exposure throughout development and early adult life to concentrations significantly higher than the Bt protein concentration likely encountered by A.custos in the field, even when feeding directly on Bt plants. Enzyme-linked immunosorbent assay confirmed the presence of Bt proteins in A. custos midguts, but quantitative real-time PCR analysis of 12 genes associated with detoxification, antioxidative responses, immune responses, and metabolism revealed no significant changes in expression in adult bugs. Indirect exposure to these toxins via consumption of intoxicated prey, larvae of Helicoverpa armigera (Hübner), likewise produced no negative impacts on survival, development, adult weight, or female fecundity in either the F0 (exposed) or F1 (unexposed) generation, but female fresh weight was reduced in the F0 generation by the Cry1Ah (50 µg/g) treatment. Finally, a competitive binding assay with labelled protein and a ligand blotting assay both demonstrated that the Cry1Ah protein could not bind to receptors on the midgut brush border membrane vesicles (BBMVs) of A. custos adults. Therefore, we conclude that Cry1Ah and Vip3Aa proteins are unlikely to have significant negative effects on A. custos populations if employed as plant-incorporated protectants in field crops.