Microbiota of pest insect Nezara viridula mediate detoxification and plant defense repression.

The Southern green shield bug, Nezara viridula, is an invasive piercing and sucking pest insect that feeds on crop plants and poses a threat to global food production. Given that insects are known to live in a close relationship with microorganisms, our study provides insights into the community composition and function of the N. viridula-associated microbiota and its effect on host-plant interactions. We discovered that N. viridula hosts both vertically and horizontally transmitted microbiota throughout different developmental stages and their salivary glands harbor a thriving microbial community that is transmitted to the plant while feeding. The N. viridula microbiota was shown to aid its host with the detoxification of a plant metabolite, namely 3-nitropropionic acid, and repression of host plant defenses. Our results demonstrate that the N. viridula-associated microbiota plays an important role in interactions between insects and plants and could therefore be considered a valuable target for the development of sustainable pest control strategies.

Trichomes and unique gene expression confer insect herbivory resistance in Vitis labrusca grapevines.

BACKGROUND: Grapevine (Vitis) is one of the world's most valuable fruit crops, but insect herbivory can decrease yields. Understanding insect herbivory resistance is critical to mitigating these losses. Vitis labrusca, a wild North American grapevine species, has been leveraged in breeding programs to generate hybrid grapevines with enhanced abiotic and biotic stress resistance, rendering it a valuable genetic resource for sustainable viticulture. This study assessed the resistance of V. labrusca acc. 'GREM4' and Vitis vinifera cv. 'PN40024' grapevines to Popillia japonica (Japanese beetle) herbivory and identified morphological and genetic adaptations underlying this putative resistance. RESULTS: 'GREM4' displayed greater resistance to beetle herbivory compared to 'PN40024' in both choice and no-choice herbivory assays spanning periods of 30 min to 19 h. 'GREM4' had significantly higher average leaf trichome densities than 'PN40024' and beetles preferred to feed on the side of leaves with fewer trichomes. When leaves from each species that specifically did not differ in trichome densities were fed on by beetles, significantly less leaf area was damaged in 'GREM4' (3.29mm2) compared to 'PN40024' (9.80mm2), suggesting additional factors beyond trichomes contributed to insect herbivory resistance in 'GREM4'. Comparative transcriptomic analyses revealed 'GREM4' exhibited greater constitutive (0 h) expression of defense response and secondary metabolite biosynthesis genes compared to 'PN40024', indicative of heightened constitutive defenses. Upon herbivory, 'GREM4' displayed a greater number of differentially expressed genes (690) compared to 'PN40024' (502), suggesting a broader response. Genes up-regulated in 'GREM4' were enriched in terpene biosynthesis, flavonoid biosynthesis, phytohormone signaling, and disease defense-related functions, likely contributing to heighted insect herbivory defense, while genes differentially expressed in 'PN40024' under herbivory were enriched in xyloglucan, cell wall formation, and calcium ion binding. The majority of genes implicated in insect herbivory defense were orthologs with specific expression patterns in 'GREM4' and 'PN40024', but some paralogous and genome-specific genes also likely contributed to conferring resistance. CONCLUSIONS: Our findings suggest that 'GREM4' insect herbivory resistance was attributed to a combination of factors, including trichomes and unique constitutive and inducible expression of genes implicated in terpene, flavonoid, and phenylpropanoid biosynthesis, as well as pathogen defense.

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.

Does spatial variation in insect herbivory match variations in plant quality? A meta-analysis.

Variation in herbivore pressure has often been predicted from patterns in plant traits considered as antiherbivore defences. Here, we tested whether spatial variation in field insect herbivory is associated with the variation in plant quality by conducting a meta-analysis of 223 correlation coefficients between herbivory levels and the expression of selected plant traits. We found no overall correlation between herbivory and either concentrations of plant secondary metabolites or values of physical leaf traits. This result was due to both the large number of low correlations and the opposing directions of high correlations in individual studies. Field herbivory demonstrated a significant association only with nitrogen: herbivore pressure increased with an increase in nitrogen concentration in plant tissues. Thus, our meta-analysis does not support either theoretical prediction, i.e., that plants possess high antiherbivore defences in localities with high herbivore pressure or that herbivory is low in localities where plant defences are high. We conclude that information about putative plant defences is insufficient to predict plant losses to insects in field conditions and that the only bottom-up factor shaping spatial variation in insect herbivory is plant nutritive value. Our findings stress the need to improve a theory linking plant putative defences and herbivory.

Contrasting defence mechanisms against spider mite infestation in cyanogenic and non-cyanogenic legumes.

Understanding the complex interactions between plants and herbivores is essential for improving crop resistance. Aiming to expand the role of cyanogenesis in plant defence, we investigated the response of the cyanogenic Phaseolus lunatus (lima bean) and the non-cyanogenic Phaseolus vulgaris (common bean) to Tetranychus urticae (spider mite) infestation. Despite mite infesting both legumes, leaf damage infringed by this feeder was reduced in lima bean. Comparative transcriptome analyses revealed that both species exhibited substantial metabolic and transcriptional changes upon infestation, although alterations in P. lunatus were significantly more pronounced. Specific differences in amino acid homeostasis and key genes associated with the cyanogenic pathway were observed in these species, as well as the upregulation of the mandelonitrile lyase gene (PlMNL1) following T. urticae feeding. Concomitantly, the PIMNL1 activity increased. Lima bean plants also displayed an induction of ß-cyanoalanine synthase (PlCYSC1), a key enzyme for cyanide detoxification, suggesting an internal regulatory mechanism to manage the toxicity of their defence responses. These findings contribute to our understanding of the legume-herbivore interactions and underscore the potential role of cyanogenesis in the elaboration of specific defensive responses, even within the same genus, which may reflect distinctive evolutionary adaptations or varying metabolic capabilities between species.

Induced resistance to herbivory and the intelligent plant.

Plant induced responses to environmental stressors are increasingly studied in a behavioral ecology context. This is particularly true for plant induced responses to herbivory that mediate direct and indirect defenses, and tolerance. These seemingly adaptive alterations of plant defense phenotypes in the context of other environmental conditions have led to the discussion of such responses as intelligent behavior. Here we consider the concept of plant intelligence and some of its predictions for chemical information transfer in plant interaction with other organisms. Within this framework, the flow, perception, integration, and storage of environmental information are considered tunable dials that allow plants to respond adaptively to attacking herbivores while integrating past experiences and environmental cues that are predictive of future conditions. The predictive value of environmental information and the costs of acting on false information are important drivers of the evolution of plant responses to herbivory. We identify integrative priming of defense responses as a mechanism that allows plants to mitigate potential costs associated with acting on false information. The priming mechanisms provide short- and long-term memory that facilitates the integration of environmental cues without imposing significant costs. Finally, we discuss the ecological and evolutionary prediction of the plant intelligence hypothesis.

Begomovirus Transmission to Tomato Plants Is Not Hampered by Plant Defenses Induced by Dicyphus hesperus Knight.

Plants can respond to insect infestation and virus infection by inducing plant defenses, generally mediated by phytohormones. Moreover, plant defenses alter host quality for insect vectors with consequences for the spread of viruses. In agricultural settings, other organisms commonly interact with plants, thereby inducing plant defenses that could affect plant-virus-vector interactions. For example, plant defenses induced by omnivorous insects can modulate insect behavior. This study focused on tomato yellow leaf curl virus (TYLCV), a plant virus of the family Geminiviridae and genus Begomovirus. It is transmitted in a persistent circulative manner by the whitefly Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae), posing a global threat to tomato production. Mirids (Hemiptera: Miridae) are effective biological control agents of B. tabaci, but there is a possibility that their omnivorous nature could also interfere with the process of virus transmission. To test this hypothesis, this study first addressed to what extent the mirid bug Dicyphus hesperus Knight induces plant defenses in tomato. Subsequently, the impact of this plant-omnivore interaction on the transmission of TYLCV was evaluated. Controlled cage experiments were performed in a greenhouse setting to evaluate the impact of mirids on virus transmission and vector acquisition by B. tabaci. While we observed a reduced number of whiteflies settling on plants exposed to D. hesperus, the plant defenses induced by the mirid bug did not affect TYLCV transmission and accumulation. Additionally, whiteflies were able to acquire comparable amounts of TYLCV on mirid-exposed plants and control plants. Overall, the induction of plant defenses by D. hesperus did not influence TYLCV transmission by whiteflies on tomato.

Defence and nutrition synergistically contribute to the distinct tolerance of rice subspecies to the stem borer, Chilo suppressalis.

Damage caused by the rice striped stem borer (SSB), Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), is much more severe on indica/xian rice than on japonica/geng rice (Oryza sativa) which matches pest outbreak data in cropping regions of China. The mechanistic basis of this difference among rice subspecies remains unclear. Using transcriptomic, metabolomic and genetic analyses in combination with insect bioassay experiments, we showed that japonica and indica rice utilise different defence responses to repel SSB, and that SSB exploited plant nutrition deficiencies in different ways in the subspecies. The more resistant japonica rice induced patterns of accumulation of methyl jasmonate (MeJA-part of a defensive pathway) and vitamin B1 (VB1-a nutrition pathway) distinct from indica cultivars. Using gene-edited rice plants and SSB bioassays, we found that MeJA and VB1 jointly affected the performance of SSB by disrupting juvenile hormone levels. In addition, genetic variants of key biosynthesis genes in the MeJA and VB1 pathways (OsJMT and OsTH1, respectively) differed between japonica and indica rice and contributed to performance differences; in indica rice, SSB avoided the MeJA defence pathway and hijacked the VB1 nutrition-related pathway to promote development. The findings highlight important genetic and mechanistic differences between rice subspecies affecting SSB damage which could be exploited in plant breeding for resistance.

The double life of trichomes: understanding their dual role in herbivory and herbicide resistance.

Understanding the evolutionary forces that maintain phenotypic variation in ecologically relevant traits has long been one of the central goals of evolutionary ecology. While the maintenance of variation in plant defense is most often hypothesized to be due to trait trade-offs or spatiotemporal variation in herbivore abundance, the role that heterogeneous selective agents may play on the maintenance of variation in plant defense is less examined. Trichomes are hair-like appendages on plant surfaces that can defend against multiple damaging agents such as pathogens, herbivores, and UV radiation. It is currently unknown however if conflicting selection from such heterogeneous agents of damage may act to maintain the variation observed in trichome traits. Here, we assess whether trichomes serve as an herbicide resistance trait and how it coincides with the conventionally studied defensive strategy of herbivory resistance. In a series of experiments, we exposed the annual invasive velvetleaf (Abutilon theophrasti) to glyphosate (active ingredient in "Roundup") to investigate whether trichome traits (type and density) are linked to herbicide resistance and to test whether herbicide influences selection on plant trichomes. We found that an increased proportion of branched trichomes positively impacted herbicide resistance and chewing herbivory resistance. We also found evidence that glyphosate imposes positive selection on branched trichomes in velvetleaf. Overall, our results indicate that branched trichomes can contribute to both herbicide and herbivory resistance, serving a concordant rather than conflicting role to reduce plant injury. Our findings further suggest that novel anthropogenic agents of selection can alter the composition of plant defense traits, potentially impacting trait-mediated interactions among external stressors.

Anti-herbivory defences delivered by Epichloë fungal endophytes: a quantitative review of alkaloid concentration variation among hosts and plant parts.

BACKGROUND AND AIMS: In the subfamily Poöideae (Poaceae), certain grass species possess anti-herbivore alkaloids synthesized by fungal endophytes that belong to the genus Epichloë (Clavicipitaceae). The protective role of these symbiotic endophytes can vary, depending on alkaloid concentrations within specific plant-endophyte associations and plant parts. METHODS: We conducted a literature review to identify articles containing alkaloid concentration data for various plant parts in six important pasture species, Lolium arundinaceum, Lolium perenne, Lolium pratense, Lolium multiflorum|Lolium rigidum and Festuca rubra, associated with their common endophytes. We considered the alkaloids lolines (1-aminopyrrolizidines), peramine (pyrrolopyrazines), ergovaline (ergot alkaloids) and lolitrem B (indole-diterpenes). While all these alkaloids have shown bioactivity against insect herbivores, ergovaline and lolitrem B are harmful for mammals. KEY RESULTS: Loline alkaloid levels were higher in the perennial grasses L. pratense and L. arundinaceum compared to the annual species L. multiflorum and L. rigidum, and higher in reproductive tissues than in vegetative structures. This is probably due to the greater biomass accumulation in perennial species that can result in higher endophyte mycelial biomass. Peramine concentrations were higher in L. perenne than in L. arundinaceum and not affected by plant part. This can be attributed to the high within-plant mobility of peramine. Ergovaline and lolitrem B, both hydrophobic compounds, were associated with plant parts where fungal mycelium is usually present, and their concentrations were higher in plant reproductive tissues. Only loline alkaloid data were sufficient for below-ground tissue analyses and concentrations were lower than in above-ground parts. CONCLUSIONS: Our study provides a comprehensive synthesis of fungal alkaloid variation across host grasses and plant parts, essential for understanding the endophyte-conferred defence extent. The patterns can be understood by considering endophyte growth within the plant and alkaloid mobility. Our study identifies research gaps, including the limited documentation of alkaloid presence in roots and the need to investigate the influence of different environmental conditions.

Electrical signalling and plant response to herbivory: A short review.

For a long time, electrical signaling was neglected at the expense of signaling studies in plants being concentrated with chemical and hydraulic signals. Studies conducted in recent years have revealed that plants are capable of emitting, processing, and transmitting bioelectrical signals to regulate a wide variety of physiological functions. Many important biological and physiological phenomena are accompanied by these cellular electrical manifestations, which supports the hypothesis about the importance of bioelectricity as a fundamental 'model' for response the stresses environmental and for activities regeneration of these organisms. Electrical signals have also been characterized and discriminated against in genetically modified plants under stress mediated by sucking insects and/or by the application of systemic insecticides. Such results can guide future studies that aim to elucidate the factors involved in the processes of resistance to stress and plant defense, thus aiding in the development of successful strategies in integrated pest management. Therefore, this mini review includes the results of studies aimed at electrical signaling in response to biotic stress. We also demonstrated how the generation and propagation of electrical signals takes place and included a description of how these electrical potentials are measured.

Plant size, latitude, and phylogeny explain within-population variability in herbivory.

Interactions between plants and herbivores are central in most ecosystems, but their strength is highly variable. The amount of variability within a system is thought to influence most aspects of plant-herbivore biology, from ecological stability to plant defense evolution. Our understanding of what influences variability, however, is limited by sparse data. We collected standardized surveys of herbivory for 503 plant species at 790 sites across 116° of latitude. With these data, we show that within-population variability in herbivory increases with latitude, decreases with plant size, and is phylogenetically structured. Differences in the magnitude of variability are thus central to how plant-herbivore biology varies across macroscale gradients. We argue that increased focus on interaction variability will advance understanding of patterns of life on Earth.

GWAS analysis of maize host plant resistance to western corn rootworm (Coleoptera: Chrysomelidae) reveals candidate small effect loci for resistance breeding.

Western corn rootworm, Diabrotica virgifera virgifera (LeConte) (Coleoptera: Chrysomelidae), is the most serious economic pest of maize, Zea mays (L.) (Poales: Poaceae), in the U.S. Corn Belt and also threatens production in Europe. Traditional management options have repeatedly failed over time as western corn rootworm rapidly develops resistance to insecticides, transgenic maize and even crop rotation. Traits that improve host plant resistance and tolerance are highly sought after by plant breeders for crop protection and pest management. However, maize resistance to western corn rootworm appears to be highly complex and despite over 75 yr of breeding efforts, there are no naturally resistant hybrids available commercially. Using phenotypic data from field and greenhouse experiments on a highly diverse collection of 282 inbred lines, we screened and genetically mapped western corn rootworm-related traits to identify genetic loci which may be useful for future breeding or genetic engineering efforts. Our results confirmed that western corn rootworm resistance is complex with relatively low heritability due in part to strong genotype by environment impacts and the inherent difficulties of phenotyping below ground root traits. The results of the Genome Wide Associated Study identified 29 loci that are potentially associated with resistance to western corn rootworm. Of these loci, 16 overlap with those found in previous transcription or mapping studies indicating a higher likelihood they are truly involved in maize western corn rootworm resistance. Taken together with previous studies, these results indicate that breeding for natural western corn rootworm resistance will likely require the stacking of multiple small effect loci.

A tripartite rheostat controls self-regulated host plant resistance to insects.

Plants deploy receptor-like kinases and nucleotide-binding leucine-rich repeat receptors to confer host plant resistance (HPR) to herbivores1. These gene-for-gene interactions between insects and their hosts have been proposed for more than 50 years2. However, the molecular and cellular mechanisms that underlie HPR have been elusive, as the identity and sensing mechanisms of insect avirulence effectors have remained unknown. Here we identify an insect salivary protein perceived by a plant immune receptor. The BPH14-interacting salivary protein (BISP) from the brown planthopper (Nilaparvata lugens Stål) is secreted into rice (Oryza sativa) during feeding. In susceptible plants, BISP targets O. satvia RLCK185 (OsRLCK185; hereafter Os is used to denote O. satvia-related proteins or genes) to suppress basal defences. In resistant plants, the nucleotide-binding leucine-rich repeat receptor BPH14 directly binds BISP to activate HPR. Constitutive activation of Bph14-mediated immunity is detrimental to plant growth and productivity. The fine-tuning of Bph14-mediated HPR is achieved through direct binding of BISP and BPH14 to the selective autophagy cargo receptor OsNBR1, which delivers BISP to OsATG8 for degradation. Autophagy therefore controls BISP levels. In Bph14 plants, autophagy restores cellular homeostasis by downregulating HPR when feeding by brown planthoppers ceases. We identify an insect saliva protein sensed by a plant immune receptor and discover a three-way interaction system that offers opportunities for developing high-yield, insect-resistant crops.

Helicoverpa zea-Associated Gut Bacteria as Drivers in Shaping Plant Anti-herbivore Defense in Tomato.

Insect-associated bacteria can mediate the intersection of insect and plant immunity. In this study, we aimed to evaluate the effects of single isolates or communities of gut-associated bacteria of Helicoverpa zea larvae on herbivore-induced defenses in tomato. We first identified bacterial isolates from the regurgitant of field-collected H. zea larvae by using a culture-dependent method and 16S rRNA gene sequencing. We identified 11 isolates belonging to the families Enterobacteriaceae, Streptococcaceae, Yersiniaceae, Erwiniaceae, and unclassified Enterobacterales. Seven different bacterial isolates, namely Enterobacteriaceae-1, Lactococcus sp., Klebsiella sp. 1, Klebsiella sp. 3, Enterobacterales, Enterobacteriaceae-2, and Pantoea sp., were selected based on their phylogenetic relationships to test their impacts on insect-induced plant defenses. We found that the laboratory population of H. zea larvae inoculated with individual isolates did not induce plant anti-herbivore defenses, whereas larvae inoculated with a bacterial community (combination of the 7 bacterial isolates) triggered increased polyphenol oxidase (PPO) activity in tomato, leading to retarded larval development. Additionally, field-collected H. zea larvae with an unaltered bacterial community in their gut stimulated higher plant defenses than the larvae with a reduced gut microbial community. In summary, our findings highlight the importance of the gut microbial community in mediating interactions between herbivores and their host plants.

Drought impairs herbivore-induced volatile terpene emissions by ponderosa pine but not through constraints on newly assimilated carbon.

Volatile terpenes serve multiple biological roles including tree resistance against herbivores. The increased frequency and severity of drought stress observed in forests across the globe may hinder trees from producing defense-related volatiles in response to biotic stress. To assess how drought-induced physiological stress alters volatile emissions alone and in combination with a biotic challenge, we monitored pre-dawn water potential, gas-exchange, needle terpene concentrations and terpene volatile emissions of ponderosa pine (Pinus ponderosa) saplings during three periods of drought and in response to simulated herbivory via methyl jasmonate application. Although 3-, 6- and 7-week drought treatments reduced net photosynthetic rates by 20, 89 and 105%, respectively, the magnitude of volatile fluxes remained generally resistant to drought. Herbivore-induced emissions, however, exhibited threshold-like behavior; saplings were unable to induce emissions above constitutive levels when pre-dawn water potentials were below the approximate zero-assimilation point. By comparing compositional shifts in emissions to needle terpene concentrations, we found evidence that drought effects on constitutive and herbivore-induced volatile flux and composition are primarily via constraints on the de novo fraction, suggesting that reduced photosynthesis during drought limits the carbon substrate available for de novo volatile synthesis. However, results from a subsequent 13CO2 pulse-chase labeling experiment then confirmed that both constitutive (<3% labeled) and herbivore-induced (<8% labeled) de novo emissions from ponderosa pine are synthesized predominantly from older carbon sources with little contribution from new photosynthates. Taken together, we provide evidence that in ponderosa pine, drought does not constrain herbivore-induced de novo emissions through substrate limitation via reduced photosynthesis, but rather through more sophisticated molecular and/or biophysical mechanisms that manifest as saplings reach the zero-assimilation point. These results highlight the importance of considering drought severity when assessing impacts on the herbivore-induced response and suggest that drought-altered volatile metabolism constrains induced emissions once a physiological threshold is surpassed.

Exogenously applied melatonin alleviates the damage in cucumber plants caused by Aphis goosypii through altering the insect behavior and inducing host plant resistance.

BACKGROUND: Aphis gossypii Glover is the main pest found in most cucumber-producing areas. Melatonin (MT) has been widely studied in protecting plants from environmental stresses and pathogens. However, little knowledge is available on the impact of MT on insect resistance. RESULTS: The fecundity of aphids on MT-treated cucumber leaves was inhibited. Interestingly, MT-treated plants were more attractive to aphids, which would prevent the large-scale transmission of viruses caused by the random movement of aphids. Meanwhile, MT caused varying degrees of change in enzyme activities related to methylesterified HG degradation, antioxidants, defense systems and membrane lipid peroxidation. Furthermore, transcriptomic analysis showed that MT induced 2360 differentially expressed genes (DEGs) compared with the control before aphid infection. These DEGs mainly were enriched in hormone signal transduction, MAPK signaling pathway, and plant-pathogen interaction, revealing that MT can help plants acquire inducible resistance and enhance plant immunity. Subsequently, 2397 DEGs were identified after aphid infection. Further analysis showed that MT-treated plants possessed stronger JA signal, reactive oxygen species stability, and the ability of flavonoid synthesis under aphid infection, while mediating plant growth and sucrose metabolism. CONCLUSION: In summary, MT as an environmentally friendly substance mitigated aphid damage to cucumbers by affecting the aphids themselves and enhancing plant resistance. This will facilitate exploring sustainable MT-based strategies for cucumber aphid control. © 2022 Society of Chemical Industry.

Silicon as resistance inducer to control black apphid Aphis craccivora Koch, 1854 in cowpea beans [Vigna unguiculata (L)] Walp

The black aphid Aphis craccivora (Koch, 1854) stands out between the bugs considered cowpea pests. The objective of this study was to evaluate the effects of silicon application on the resistance induction of cowpea plants to the black aphid A. craccivora. The experiment was conducted in the Entomology Laboratory of the Phytosanitary sector of the Centro de Ciências Agrárias at the Universidade Federal do Piauí. The effects of the application of silicon on biological aspects were evaluated using a completely randomized design, with four treatments and 40 repetitions, being: silicon in soil (T1), silicon in soil + leaf (T2), silicone leaf (T3), and control (T4). The following biological variables were evaluated: generation period, reproductive period, fecundity, and daily average of nymphs per female. The silicon and lignin contents were also evaluated in the plants. The silicic acid was applied in a 1% solution around the stem of the plants (soil), 15 days after emergence, by diluting 2 g of the product in 200 mL of water. However, the leaf application was carried out with sprayer five days after application in soil. The non-preference of A. craccivora on bean was also evaluated. The evaluations were performed after 24, 48 and 72 hours of infestation by counting nymphs at 24, 48 and 72 hours and adults at each leaf session. The application of silicon promotes the reduction of the production of nymphs, interfering in the biological aspects of A. craccivora, and has potential to be used in a cowpea pest management program in cowpea.

Which variables influence the herbivory amount on Montrichardia spp. (Araceae) in aquatic ecosystems? / ¿Qué variables influyen en la herbivoría en Montrichardia spp. (Araceae) en los ecosistemas acuáticos?

Abstract Introduction: The study of herbivory is fundamental in ecology and includes how plants invest in strategies and mechanisms to reduce herbivore damage. However, there is still a lack of information about how the environment, plant density, and functional traits influence herbivory in aquatic ecosystems. Objective: To assess if there is a relationship between herbivory, environmental variables, and plant traits two species of Montrichardia, a neotropical aquatic plant. Methods: In September 2018, we studied 78 specimens of Montrichardia arborescens and 18 of Montrichardia linifera, in 18 sites in Melgaço, Pará, Brazil. On each site, we measured water depth, distance to the margin, and plant density. From plants, we measured plant height and leaf thickness, and photographed the leaves to calculate the specific leaf area and percentage herbivory. To identify anatomical structures, we collected fully expanded leaves from three individuals per quadrat. Results: For M. arborescens, plants with thicker leaves and higher specific leaf area have less herbivore damage. For M. linifera, plants from deeper sites and with thicker leaves had more herbivore damage, while plants that grew farther from the margin had less damage. We found anatomical structures related to defense, such as idioblast cells with phenolic compounds, and cells with solid inclusions that can contribute to avoiding severe damage. Conclusions: Herbivory in these Montrichardia species can be explained by a combination of plant and environmental traits (patch isolation and water depth). The main plant traits are leaf thickness and area, but chemical compounds and solid inclusions also help Montrichardia to sustain less damage than other macrophytes.

Resumen Introducción: La herbivoría es fundamental para comprender cómo las plantas invierten en diferentes estrategias para evitar la depredación, lo que implica diferentes mecanismos de defensa. Factores relacionados con el medio ambiente, la densidad de plantas y/o los rasgos funcionales de las plantas pueden influir en la herbivoría en los ecosistemas acuáticos. Sin embargo, todavía falta información sobre cómo esos factores influyen en la herbivoría en los ecosistemas acuáticos y pueden contribuir a la carga de herbivoría. Objetivo: Evaluar si existe una relación entre la herbivoría y las variables ambientales (p. ej., profundidad del agua y distancia al margen), los factores ecológicos (densidad de plantas) y los rasgos estructurales de las plantas (altura, grosor de la hoja y área foliar) e indicar estructuras anatómicas que actúen junto con los rasgos estructurales en el sistema de defensa de especies de Montrichardia. Métodos: Se evaluaron 96 individuos de Montrichardia spp. (78 de M. arborescens y 18 de M. linifera, en 18 sitios) recolectados en septiembre de 2018. En cada sitio, se midió la profundidad del agua, la distancia al margen y la densidad de plantas. De los individuos, medimos la altura de la planta, el grosor de la hoja y fotografiamos las hojas para calcular el área foliar específica y la cantidad de herbivoría (en porcentaje). Para identificar las estructuras anatómicas relacionadas con la defensa de las plantas, se recogió hojas completamente expandidas de tres individuos por cuadrante. Resultados: Para M. arborescens, las plantas con hojas más gruesas y mayor área foliar específica tienen menos daño por herbivoría. Para M. linifera, las plantas con hojas más gruesas y que habitan en sitios más profundos tienen más daño por herbívoros, mientras que las plantas más alejadas del margen tienen menos daño por herbívoros. Se encontró estructuras anatómicas relacionadas con la defensa, como células idioblásticas con compuestos fenólicos y células con inclusiones sólidas que pueden contribuir a evitar daños severos en las características de las hojas. Conclusiones: Nuestros resultados indican que la herbivoría en las especies de Montrichardia podría explicarse por una combinación de características ambientales (aislamiento del parche y profundidad del agua) y de la planta. Descubrimos que los rasgos de las hojas eran factores importantes que impulsaban los cambios en la carga de herbivoría, especialmente el grosor de las hojas y el área foliar específica. Además, las especies de Montrichardia invierten en compuestos químicos e inclusiones sólidas para evitar daños graves en las hojas y, por lo tanto, pueden sufrir menos daños que otras especies de macrófitos.

Functional evidence supports adaptive plant chemical defense along a geographical cline.

Environmental clines in organismal defensive traits are usually attributed to stronger selection by enemies at lower latitudes or near the host's range center. Nonetheless, little functional evidence has supported this hypothesis, especially for coevolving plants and herbivores. We quantified cardenolide toxins in seeds of 24 populations of common milkweed (Asclepias syriaca) across 13 degrees of latitude, revealing a pattern of increasing cardenolide concentrations toward the host's range center. The unusual nitrogen-containing cardenolide labriformin was an exception and peaked at higher latitudes. Milkweed seeds are eaten by specialist lygaeid bugs that are even more tolerant of cardenolides than the monarch butterfly, concentrating most cardenolides (but not labriformin) from seeds into their bodies. Accordingly, whether cardenolides defend seeds against these specialist bugs is unclear. We demonstrate that Oncopeltus fasciatus (Lygaeidae) metabolized two major compounds (glycosylated aspecioside and labriformin) into distinct products that were sequestered without impairing growth. We next tested several isolated cardenolides in vitro on the physiological target of cardenolides (Na+/K+-ATPase); there was little variation among compounds in inhibition of an unadapted Na+/K+-ATPase, but tremendous variation in impacts on that of monarchs and Oncopeltus. Labriformin was the most inhibitive compound tested for both insects, but Oncopeltus had the greater advantage over monarchs in tolerating labriformin compared to other compounds. Three metabolized (and stored) cardenolides were less toxic than their parent compounds found in seeds. Our results suggest that a potent plant defense is evolving by natural selection along a geographical cline and targets specialist herbivores, but is met by insect tolerance, detoxification, and sequestration.