Effects of climate change on Lepidoptera pollen loads and their pollination services in space and time.

Shifts in flowering time among plant communities as a result of climate change, including extreme weather events, are a growing concern. These plant phenological changes may affect the quantity and quality of food sources for specialized insect pollinators. Plant-pollinator interactions are threatened by habitat alterations and biodiversity loss, and changes in these interactions may lead to declines in flower visitors and pollination services. Most prior research has focused on short-term plant-pollinator interactions, which do not accurately capture changes in pollination services. Here, we characterized long-term plant-pollinator interactions and identified potential risks to specialized butterfly species due to habitat loss, fragmented landscapes, and changes in plant assemblages. We used 21 years of historical data from museum specimens to track the potential effects of direct and indirect changes in precipitation, temperature, monsoons, and wildfires on plant-pollinator mutualism in the Great Basin and Sierra Nevada. We found decreased pollen richness associated with butterflies within sites, as well as an increase in pollen grain abundance of drought-tolerant plants, particularly in the past 10 years. Moreover, increased global temperatures and the intensity and frequency of precipitation and wildfires were negatively correlated with pollen diversity. Our findings have important implications for understanding plant-pollinator interactions and the pollination services affected by global warming.

Identification of the female sex pheromone of Bastilla arctotaenia (Lepidoptera: Erebidae).

(3Z,6Z,9Z)-3,6,9-henicosatriene was identified as a major component of female sex pheromone of Bastilla arctotaenia (Lepidoptera: Erebidae), a pest of cultivated roses, by gas chromatograph-electroantennographic detector( GC-EAD) and gas chromatograph/mass spectrometry (GC/MS) analyses. The single (3Z,6Z,9Z)-3,6,9-henicosatriene (1.0 mg/lure) successfully attracted B. arctotaenia males in the field.

Phenotypic plasticity in plant defense across life stages: Inducibility, transgenerational induction, and transgenerational priming in wild radish.

As they develop, many plants deploy shifts in antiherbivore defense allocation due to changing costs and benefits of their defensive traits. Plant defenses are known to be primed or directly induced by herbivore damage within generations and across generations by long-lasting epigenetic mechanisms. However, little is known about the differences between life stages of epigenetically inducible defensive traits across generations. To help fill this knowledge gap, we conducted a multigenerational experiment to determine whether defense induction in wild radish plants was reflected in chromatin modifications (DNA methylation); we then examined differences between seedlings and reproductive plants in current and transgenerational plasticity in chemical (glucosinolates) and physical (trichomes) defenses in this species. Herbivory triggered genome methylation both in targeted plants and their offspring. Within one generation, both defenses were highly inducible at the seedling stage, but only chemical defenses were inducible in reproductive plants. Across generations, herbivory experienced by mother plants caused strong direct induction of physical defenses in their progeny, with effects lasting from seedling to reproductive stages. For chemical defenses, however, this transgenerational induction was evident only in adults. Transgenerational priming was observed in physical and chemical defenses, particularly in adult plants. Our results show that transgenerational plasticity in plant defenses in response to herbivore offense differs for physical and chemical defense and changes across plant life stages.

Better Together: Volatile-Mediated Intraguild Effects on the Preference of Tuta absoluta and Trialeurodes vaporariorum for Tomato Plants.

Plant-herbivore interactions have been extensively studied in tomato plants and their most common pests. Tomato plant chemical defenses, both constitutive and inducible, play a role in mediating these interactions. Damaged tomato plants alter their volatile profiles, affecting herbivore preferences between undamaged and damaged plants. However, previous studies on tomato volatiles and herbivore preferences have yielded conflicting results, both in the volatile chemistry itself as well as in the attraction/repellent herbivore response. This study revisits the volatile-mediated interactions between tomato plants and two of their main herbivores: the leafminer Tuta absoluta and the whitefly Trialeurodes vaporariorum. Tomato plant volatiles were analyzed before and after damage by each of these herbivores, and the preference for oviposition (T. absoluta) and settling (T. vaporariorum) on undamaged and damaged plants was assessed both after conspecific and heterospecific damage. We found that both insects consistently preferred damaged plants over undamaged plants. The emission of herbivore-induced plant volatiles (HIPVs) increased after T. absoluta damage but decreased after T. vaporariorum damage. While some of our findings are in line with previous reports, T. absoluta preferred to oviposit on plants damaged by conspecifics, which differs from earlier studies. A comparison of HIPVs emitted after damage by T. absoluta and T. vaporariorum revealed differences in up- or down-regulation, as well as significant variations in specific compounds (12 for T. absoluta and 26 for T. vaporariorum damaged-plants). Only two compounds, ß-caryophyllene and tetradecane, significantly varied because of damage by either herbivore, in line with the overall variation of the HIPV blend. Differences in HIPVs and herbivore preferences may be attributed to the distinct feeding habits of both herbivores, which activate different defensive pathways in plants. The plant's challenge in simultaneously activating both defensive pathways may explain the preference for heterospecific damaged plants found in this study, which are also in line with our own observations in greenhouses.

Generalists are more specialized in low-resource habitats, increasing stability of ecological network structure.

Linking mechanistic processes to the stability of ecological networks is a key frontier in ecology. In trophic networks, "modules"-groups of species that interact more with each other than with other members of the community-confer stability, mitigating effects of species loss or perturbation. Modularity, in turn, is shaped by the interplay between species' diet breadth traits and environmental influences, which together dictate interaction structure. Despite the importance of network modularity, variation in this emergent property is poorly understood in complex natural systems. Using two years of field data, we quantified interactions between a rich community of lepidopteran herbivores and their host plants across a mosaic of low-resource serpentine and high-resource nonserpentine soils. We used literature and our own observations to categorize herbivore species as generalists (feeding on more than one plant family) or specialists (feeding on one plant family). In both years, the plant-herbivore network was more modular on serpentine than on nonserpentine soils-despite large differences in herbivore assemblage size across years. This structural outcome was primarily driven by reduction in the breadth of host plant use by generalist species, rather than by changes in the composition of species with different fundamental diet breadths. Greater modularity-and thus greater stability-reflects environmental conditions and plastic responses by generalist herbivores to low host plant quality. By considering the dual roles of species traits and ecological processes, we provide a deeper mechanistic understanding of network modularity, and suggest a role for resource availability in shaping network persistence.

Comparative analyses of responses to exogenous and endogenous antiherbivore elicitors enable a forward genetics approach to identify maize gene candidates mediating sensitivity to herbivore-associated molecular patterns.

Crop damage by herbivorous insects remains a significant contributor to annual yield reductions. Following attack, maize (Zea mays) responds to herbivore-associated molecular patterns (HAMPs) and damage-associated molecular patterns (DAMPs), activating dynamic direct and indirect antiherbivore defense responses. To define underlying signaling processes, comparative analyses between plant elicitor peptide (Pep) DAMPs and fatty acid-amino acid conjugate (FAC) HAMPs were conducted. RNA sequencing analysis of early transcriptional changes following Pep and FAC treatments revealed quantitative differences in the strength of response yet a high degree of qualitative similarity, providing evidence for shared signaling pathways. In further comparisons of FAC and Pep responses across diverse maize inbred lines, we identified Mo17 as part of a small subset of lines displaying selective FAC insensitivity. Genetic mapping for FAC sensitivity using the intermated B73 × Mo17 population identified a single locus on chromosome 4 associated with FAC sensitivity. Pursuit of multiple fine-mapping approaches further narrowed the locus to 19 candidate genes. The top candidate gene identified, termed FAC SENSITIVITY ASSOCIATED (ZmFACS), encodes a leucine-rich repeat receptor-like kinase (LRR-RLK) that belongs to the same family as a rice (Oryza sativa) receptor gene previously associated with the activation of induced responses to diverse Lepidoptera. Consistent with reduced sensitivity, ZmFACS expression was significantly lower in Mo17 as compared to B73. Transient heterologous expression of ZmFACS in Nicotiana benthamiana resulted in a significantly increased FAC-elicited response. Together, our results provide useful resources for studying early elicitor-induced antiherbivore responses in maize and approaches to discover gene candidates underlying HAMP sensitivity in grain crops.

Baculoviruses hijack the visual perception of their caterpillar hosts to induce climbing behaviour thus promoting virus dispersal.

Baculoviruses can induce climbing behaviour in their caterpillar hosts to ensure they die at elevated positions to enhance virus transmission, providing an excellent model to study parasitic manipulation of host behaviour. Here, we demonstrate that climbing behaviour occurred mostly during daylight hours, and that the height at death of Helicoverpa armigera single nucleopolyhedrovirus (HearNPV)-infected larvae increases with the height of the light source. Phototaxic and electroretinogram (ERG) responses were enhanced after HearNPV-infection in host larvae, and ablation of stemmata in infected larvae prevented both phototaxis and climbing behaviour. Through transcriptome and quantitative PCR, we confirmed that two opsin genes (a blue light-sensitive gene, HaBL; and a long wave-sensitive gene, HaLW) as well as the TRPL (transient receptor potential-like channel protein) gene, all integral to the host's visual perception pathway, were significantly upregulated after HearNPV infection. Knockout of HaBL, HaLW, or TRPL genes using the CRISPR/Cas9 system resulted in significantly reduced ERG responses, phototaxis, and climbing behaviour in HearNPV-infected larvae. These results reveal that HearNPV alters the expression of specific genes to hijack host visual perception at fundamental levels-photoreception and phototransduction-in order to induce climbing behaviour in host larvae.

Insect-damaged Arabidopsis moves like wounded Mimosa pudica.

Slow wave potentials (SWPs) are damage-induced electrical signals which, based on experiments in which organs are burned, have been linked to rapid increases in leaf or stem thickness. The possibility that pressure surges in injured xylem underlie these events has been evoked frequently. We sought evidence for insect feeding-induced positive pressure changes in the petioles of Arabidopsis thaliana Instead, we found that petiole surfaces of leaves distal to insect-feeding sites subsided. We also found that insect damage induced longer-duration downward leaf movements in undamaged leaves. The transient petiole deformations were contemporary with and dependent on the SWP. We then investigated if mutants that affect the xylem, which has been implicated in SWP transmission, might modify SWP architecture. irregular xylem mutants strongly affected SWP velocity and kinetics and, in parallel, restructured insect damage-induced petiole deformations. Together, with force change measurements on the primary vein, the results suggest that extravascular water fluxes accompany the SWP. Moreover, petiole deformations in Arabidopsis mimic parts of the spectacular distal leaf collapse phase seen in wounded Mimosa pudica We genetically link electrical signals to organ movement and deformation and suggest an evolutionary origin of the large leaf movements seen in wounded Mimosa.

Isolation, screening and identification of key components having intense insect repellent activity against Plodia interpunctella from four different medicinal plant materials.

BACKGROUND: Global warming and the indiscriminate use of pesticides have increased the propagation of the stored-product insect pests, leading to enormous losses in the agriculture and food industries. The most used insect repellents are synthetic derivatives; however, these have an adverse effect on human health as well as on the environment. Therefore, we attempted to find materials with insect repellent activity in natural products. The present study aimed to identify the single chemical component with intense insect repellent activity in extracts from four different Oriental medicinal plant materials: (i) Anethum graveolens L. (dill) seeds; (ii) Artemisia capillaris Thunb. (capillary wormwood) leaves; (iii) smoked Prunus mume Siebold & Zucc. (mume) fruits; and (iv) Rhus javanica L. (galls). RESULTS: As a result of the bioassay-guided fractionation of each extract against the Plodia interpunctella, stored-product insect, the n-hexane fraction of dill seeds extract was confirmed as the optimal fraction between all of the fractions. In total, 32 chemical components were identified from the n-hexane fraction of dill seeds by gas chromatography-mass spectrometry analysis, and the two main components were dillapiole (47.51%) and carvone (26.76%). Of the two components, dillapiole was confirmed as the key component playing an essential role in insect repellent activity. CONCLUSION: Our study suggests that dillapiole has the potential to be used as a natural insect repellent for the control of P. interpunctella infestation in agricultural and food products during distribution and storage. © 2021 Society of Chemical Industry.

High-Throughput Feeding Bioassay for Lepidoptera Larvae.

Finding plant cultivars that are resistant or tolerant against lepidopteran pests, takes time, effort and is costly. We present here a high throughput leaf-disk consumption assay system, to screen plants for resistance or chemicals for their deterrence. A webcam capturing images at regular intervals can follow the feeding activities of 150 larvae placed into individual cages. We developed a computer program running under an open source image analysis program to analyze and measure the surface of each leaf disk over time. We further developed new statistical procedures to analyze the time course of the feeding activities of the larvae and to compare them between treatments. As a test case, we compared how European corn borer larvae respond to a commercial antifeedant containing azadirachtin, and to quinine, which is a bitter alkaloid for many organisms. As expected, increasing doses of azadirachtin reduced and delayed feeding. However, quinine was poorly effective at the range of concentrations tested (10-5 M to 10-2 M). The model cage, the camera holder, the plugins, and the R scripts are freely available, and can be modified according to the users' needs.

Triterpenes induced by young apple fruits in response to herbivore attack.

Apples Malus domestica, known as a rich source of triterpene acids, induced more variety and quantity of triterpene acids in response to herbivory or mechanical damage. There were 3 major induced compounds: pomaceic acid and euscaphic acid, both of which are known apple triterpene acids, and 2α,19α-dihydroxy-3-oxours-12-en-28-oic acid (named eriobotoric acid), which was first identified in apples. In this study, the 3 compounds' induction curves after damage, varietal differences in induction amounts, and physiological roles against pest insects were further investigated. Eriobotoric acid showed clear antifeedant activity against lepidopteran insect Spodoptera litura but not against apple pests.

Plasticity of cold hardiness in the eastern spruce budworm, Choristoneura fumiferana.

High latitude insect populations must cope with extreme conditions, particularly low temperatures. Insects use a variety of cold hardiness mechanisms to withstand this temperature stress, and these can drive geographic distributions through overwintering mortality. The degree of cold hardiness can be altered by two evolved responses: phenotypic plasticity and local adaptation. Phenotypic plasticity can occur within or between generations (transgenerational plasticity; TGP), and local adaptation can evolve through directional selection in response to regional climatic differences. We used the eastern spruce budworm, Choristoneura fumiferana (Lepidoptera: Tortricidae) as a model to explore the role that variable winter temperatures play in inducing two aspects of plasticity in cold hardiness: TGP and local adaptation in phenotypic plasticity. This species is one of the most destructive boreal forest pests in North America, therefore accurately predicting overwintering survival is essential for effective management. While we found no evidence of TGP in cold hardiness, there was a long term fitness cost to larvae that experienced repeated cold exposures. We also found evidence of local adaptation in both seasonal and short-term plasticity of cold hardiness, as our more northerly populations that would experience lower overwintering temperatures had more plastic responses to cold exposure. These findings provide evidence for the importance of phenotypic plasticity and local adaptation when modelling species distributions.

Effects of brief exposure to high temperatures on the survival and fecundity of Athetis lepigone (Lepidoptera: Noctuidae).

In Huang-Huai-Hai Summer Corn Region of China, brief periods of high summer temperatures have been reported with increasing frequency in recent years. Athetis lepigone is a cosmopolitan insect which causes severe damage on summer corn seedlings. To understand how high summer temperatures may affect the population dynamics of A. lepigone, we exposed different developmental stages (1, 2 and 4-day old eggs; 1, 6, 12 and 18-day old larvae; 1, 3 and 6-day old pupae; and 1 and 2-day old female and male adults) to 41 °C for periods of various length (0.5, 1, 2, 4 and 6 h): The rearing temperature (constant 26 °C) was used as control. After heat treatment, all individuals were transferred to a 26 °C climate chamber for further development. The effects on immediate survival, maturation success to adulthood, and female fecundity were studied. Eggs, young larvae, late pupae and newly emerged adults had relatively higher immediate survival rates than the other experimental groups. Heat treatment at the egg and larval stages had no impact on development to adulthood and on female fecundity, while it significantly reduced the survival rate of larvae but not of eggs. Brief exposure to high temperature during the early pupal stage and as adults depressed female fecundity whereas exposure during the late pupal stage had no effect.

Domestication of Plants of Ugni molinae Turcz (Myrtaceae) Interferes in the Biology of Chilesia rudis (Lepidoptera: Erebidae) Larvae.

In terms of the domestication process in murtilla, studies have found changes in the concentration of phenolic compounds, with reduction of chemical defense of plants, depending on the change in the feeding behavior of insects. Thus, we hypothesized that the domestication of Ugni molinae decreases the content of phenolic compounds and modifies the feeding preference of Chilesia rudis larvae. Leaves of three parental ecotypes and four cultivated ecotypes were used in preference experiments to evaluate the mass gain and leaves consumption of larvae. Phenolic extracts from leaves of U. molinae were analyzed by HPLC. Identified compounds were incorporated in an artificial diet to assess their effect on mass gain, consumption, and survival of the larvae. The presence of phenolic compounds in bodies and feces was also evaluated. In terms of choice assays, larvae preferred parental ecotypes. Regarding compounds, vanillin was the most varied between the ecotypes in leaves. However, plant domestication did not show a reduction in phenolic compound concentration of the ecotypes studied. Furthermore, there was no clear relation between phenolic compounds and the performance of C. rudis larvae. Whether this was because of sequestration of some compounds by larvae is unknown. Finally, results of this study could also suggest that studied phenolic compounds have no role in the C. rudis larvae resistance in this stage of murtilla domestication process.

Hybridization and introgression between Helicoverpa armigera and H. zea: an adaptational bridge.

BACKGROUND: Invasion of organisms into new ecosystems is increasingly common, due to the global trade in commodities. One of the most complex post-invasion scenarios occurs when an invasive species is related to a native pest, and even more so when they can hybridize and produce fertile progeny. The global pest Helicoverpa armigera was first detected in Brazil in 2013 and generated a wave of speculations about the possibility of hybridization with the native sister taxon Helicoverpa zea. In the present study, we used genome-wide single nucleotide polymorphisms from field-collected individuals to estimate hybridization between H. armigera and H. zea in different Brazilian agricultural landscapes. RESULTS: The frequency of hybridization varied from 15 to 30% depending on the statistical analyses. These methods showed more congruence in estimating that hybrids contained approximately 10% mixed ancestry (i.e. introgression) from either species. Hybridization also varied considerably depending on the geographic locations where the sample was collected, forming a 'mosaic' hybrid zone where introgression may be facilitated by environmental and landscape variables. Both landscape composition and bioclimatic variables indicated that maize and soybean cropland are the main factors responsible for high levels of introgression in agricultural landscapes. The impact of multiple H. armigera incursions is reflected in the structured and inbred pattern of genetic diversity. CONCLUSIONS: Our data showed that the landscape composition and bioclimatic variables influence the introgression rate between H. armigera and H. zea in agricultural areas. Continuous monitoring of the hybridization process in the field is necessary, since agricultural expansion, climatic fluctuations, changing composition of crop species and varieties, and dynamic planting seasons are some factors in South America that could cause a sudden alteration in the introgression rate between Helicoverpa species. Introgression between invasive and native pests can dramatically impact the evolution of host ranges and resistance management.

False holes as camouflage.

Long noted by naturalists, leaf mimicry provides some of the most impressive examples of camouflage through masquerade. Many species of leaf-mimicking Lepidoptera also sport wing markings that closely resemble irregularly shaped holes caused by decay or insect damage. Despite proposals that such markings can either enhance resemblance to damaged leaves or act to disrupt surface appearance through false depth cues, to our knowledge, no attempt has been made to establish exactly how these markings function, or even whether they confer a survival benefit to prey. Here, in two field experiments using artificial butterfly-like targets, we show that false hole markings provide significant survival benefits against avian predation. Furthermore, in a computer-based visual search experiment, we demonstrate that detection of such targets by humans is impeded in a similar fashion. Equally contrasting light marks do not have the same effect; indeed, they lead to increased detection. We conclude that the mechanism is the disruption of the otherwise homogeneous wing surface (surface disruptive camouflage) and that, by resembling the holes sometimes found in real leaves, the disruptive benefits are not offset by conspicuousness costs.

Towards unravelling Wolbachia global exchange: a contribution from the Bicyclus and Mylothris butterflies in the Afrotropics.

BACKGROUND: Phylogenetically closely related strains of maternally inherited endosymbiotic bacteria are often found in phylogenetically divergent, and geographically distant insect host species. The interspecies transfer of the symbiont Wolbachia has been thought to have occurred repeatedly, facilitating its observed global pandemic. Few ecological interactions have been proposed as potential routes for the horizontal transfer of Wolbachia within natural insect communities. These routes are however likely to act only at the local scale, but how they may support the global distribution of some Wolbachia strains remains unclear. RESULTS: Here, we characterize the Wolbachia diversity in butterflies from the tropical forest regions of central Africa to discuss transfer at both local and global scales. We show that numerous species from both the Mylothris (family Pieridae) and Bicyclus (family Nymphalidae) butterfly genera are infected with similar Wolbachia strains, despite only minor interclade contacts across the life cycles of the species within their partially overlapping ecological niches. The phylogenetic distance and differences in resource use between these genera rule out the role of ancestry, hybridization, and shared host-plants in the interspecies transfer of the symbiont. Furthermore, we could not identify any shared ecological factors to explain the presence of the strains in other arthropod species from other habitats, or even ecoregions. CONCLUSION: Only the systematic surveys of the Wolbachia strains from entire species communities may offer the material currently lacking for understanding how Wolbachia may transfer between highly different and unrelated hosts, as well as across environmental scales.

Evolutionary constraints on flicker fusion frequency in Lepidoptera.

Flying insects occupy both diurnal and nocturnal niches, and their visual systems encounter distinct challenges in both conditions. Visual adaptations, such as superposition eyes of moths, enhance sensitivity to low light levels but trade off with spatial and temporal resolution. Conversely, apposition eyes of butterflies enable high spatial resolution but are poorly sensitive in dim light. Although diel activity patterns of insects influence visual processing, their role in evolution of visual systems is relatively unexplored. Lepidopteran insects present an excellent system to study how diel activity patterns and phylogenetic position influence the visual transduction system. We addressed this question by comparing electroretinography measurements of temporal response profiles of diverse Lepidoptera to light stimuli that were flickering at different frequencies. Our data show that the eyes of diurnal butterflies are sensitive to visual stimuli of higher temporal frequencies than nocturnal moths. Hesperiid skippers, which are typically diurnal or crepuscular, exhibit intermediate phenotypes with peak sensitivity across broader frequency range. Across all groups, species within families exhibited similar phenotypes irrespective of diel activity. Thus, Lepidopteran photoreceptors may have diversified under phylogenetic constraints, and shifts in their sensitivity to higher temporal frequencies occurred concomitantly with the evolution of diurnal lifestyles.

3,7-Dimethylpentadecane: a Novel Sex Pheromone Component from Leucoptera sinuella (Lepidoptera: Lyonetiidae).

Leucoptera sinuella is a leaf-miner moth present in several regions in the world, which has been recently introduced into Chile. The larvae feed exclusively on the leaves of poplar and willow trees, and the damage caused by the feeding behavior poses a threat to the wood-producing industry. Besides, L. sinuella larvae invade nearby orchards for pupation, causing rejections in Chilean fresh fruit for export. Here we report the identification of the female-produced sex pheromone of L. sinuella as a first step towards the development of pheromone-based methods for pest management of this species. First, we analyzed hexane extracts of the abdominal glands of virgin females by gas chromatography coupled with mass spectrometry and identified the major compound in these extracts to be 3,7-dimethylpentadecane, while minor compounds in the extracts proved to be 3,7-dimethyltetradecane and 7-methylpentadecane. Structure assignments were carried out by comparison of retention times and mass spectra of the natural products with those of authentic reference samples. Second, we conducted field tests, which showed that traps baited with synthetic 3,7-dimethylpentadecane were significantly attractive to males in a dose-dependent response. Our results also showed that a mixture of 3,7-dimethylpentadecane, 3,7-dimethyltetradecane, and 7-methylpentadecane in proportions similar to those found in gland extracts was the most attractive lure.

Asymmetric Responses to Climate Change: Temperature Differentially Alters Herbivore Salivary Elicitor and Host Plant Responses to Herbivory.

The effect of temperature on insect-plant interactions in the face of changing climate is complex as the plant, its herbivores and their interactions are usually affected differentially leading to an asymmetry in response. Using experimental warming and a combination of biochemical and herbivory bioassays, the effects of elevated temperatures and herbivore damage (Helicoverpa zea) on resistance and tolerance traits of Solanum lycopersicum var. Better boy (tomato), as well as herbivory performance and salivary defense elicitors were examined. Insects and plants were differentially sensitive towards warming within the experimental temperature range. Herbivore growth rate increased with temperature, whereas plants growth as well as the ability to tolerate stress measured by photosynthesis recovery and regrowth ability were compromised at the highest temperature regime. In particular, temperature influenced the caterpillars' capacity to induce plant defenses due to changes in the amount of a salivary defense elicitor, glucose oxidase (GOX). This was further complexed by the temperature effects on plant inducibility, which was significantly enhanced at an above-optimum temperature; this paralleled with an increased plants resistance to herbivory but significantly varied between previously damaged and undamaged leaves. Elevated temperatures produced asymmetry in species' responses and changes in the relationship among species, indicating a more complicated response under a climate change scenario.