Generalist Herbivorous Insect Research Articles

Active defense strategies for invasive plants may alter the distribution pattern of pests in the invaded area.

In the invaded areas, it is believed that invasive species reduce their investment in defense due to the absence of natural enemies. By field investigation and a series of laboratory assays, This study explored the defense strategies of invasive plants. Field investigation indicated that invasive plants have a antifeedant effect on herbivorous pests, and the distribution frequency of wormholes of native plants shows a peak at a distance of 2-3 m from the invasive species. The feeding preference experiment conducted with two generalist herbivorous insects (native insect Spodoptera litura and invasive insect Spodoptera frugiperda) showed that the invasive plants have a stronger antifeedant effect than native plants. By analyzing the content of secondary metabolites in the leaves of three invasive plants (Sphagneticola trilobata, Mikania micrantha, Ipomoea cairica) and three native plants (Ipomoea nil, Paederia foetida, Polygonum chinense), the leaves of invasive plants had higher concentrations of substances associated with defenses, including total phenols, flavonoids, jasmonic acid, tannin, H2O2, and total antioxidant capacity (TAC), and lower soluble protein concentrations than native plants. After leaf damage, compared to native plants, the leaves of invasive plants showed an overall increase in substances associated with defense, except for soluble sugar. These results suggest that invasive plants maintain active defense strategies in invaded areas, leading to changes in the distribution patterns of herbivorous insects in a manner that facilitates invasion.

Host-specific gene expression as a tool for introduction success in Naupactus parthenogenetic weevils.

Food resource access can mediate establishment success in invasive species, and generalist herbivorous insects are thought to rely on mechanisms of transcriptional plasticity to respond to dietary variation. While asexually reproducing invasives typically have low genetic variation, the twofold reproductive capacity of asexual organisms is a marked advantage for colonization. We studied host-related transcriptional acclimation in parthenogenetic, invasive, and polyphagous weevils: Naupactus cervinus and N. leucoloma. We analyzed patterns of gene expression in three gene categories that can mediate weevil-host plant interactions through identification of suitable host plants, short-term acclimation to host plant defenses, and long-term adaptation to host plant defenses and their pathogens. This approach employed comparative transcriptomic methods to investigate differentially expressed host detection, detoxification, immune defense genes, and pathway-level gene set enrichment. Our results show that weevil gene expression responses can be host plant-specific, and that elements of that response can be maintained in the offspring. Some host plant groups, such as legumes, appear to be more taxing as they elicit a complex gene expression response which is both strong in intensity and specific in identity. However, the weevil response to taxing host plants shares many differentially expressed genes with other stressful situations, such as host plant cultivation conditions and transition to novel host, suggesting that there is an evolutionarily favorable shared gene expression regime for responding to different types of stressful situations. Modulating gene expression in the absence of other avenues for phenotypic adaptation may be an important mechanism of successful colonization for these introduced insects.

A sensitive period for the induction of host plant preference in a generalist herbivorous insect

Many species use early life sensory experiences to guide phenotypic plasticity and facilitate decision making during later adult settlement in new habitats or location of suitable resources. However, only some of the available sensory information in the environment is relevant for future decision making and, in addition, it may only be informative during certain times during development. This means that sampling of environmental information that should be retained to adulthood could be limited to certain sensitive periods. In the polyphagous moth Spodoptera littoralis , recent studies have demonstrated that adult females exhibit an innate preference hierarchy for certain plants that can be modulated by olfactory experience during the larval stage. This olfactory-driven phenotypic plasticity requires that information about plant quality is transferred over the full metamorphosis. However, the timing of information acquisition that affects adult behaviour and whether there is a sensitive period for the sensory experience inducing host plant preferences are not known. In this study, we tested whether experience used by adult females during host plant choice was acquired during early or later larval stages and whether adult females can retain larval experience from only one or from sequential feeding events on different plants. We found that only plant feeding experience during the later larval stages induced a host plant preference shift in adult females. Furthermore, our experiments demonstrated that when given an artificial diet in the later larval stage, adult females relied on their innate plant preference hierarchy even if they had feeding experience from a host plant in the early stages. The existence of a sensitive period during late larval stages may be constrained not only by neural development or limitations, but also by ecological factors affecting when the larval experience is most reliable for adult decisions. • Larval experience affects adult moth host plant decisions. • There is a sensitive period for host plant cue transfer over the full metamorphosis. • Only experience at late larval instars affects adult host plant choice decisions. • Adults use innate preference when exposed to early instar plant experience.

Herbivore accumulation on invasive alien plants increases the distribution range of generalist herbivorous insects and supports proliferation of non-native insect pests

Invasive alien plant species have become dominant components of many landscapes, where they are indicators of ecological disequilibria. They coexist and compete with native plants, disrupting a wide range of trophic interactions, and must cope with a new array of herbivores. We examined factors influencing the occurrence of invertebrate herbivores on two invasive alien plant species, Acacia dealbata and Carpobrotus edulis, and surrounding native vegetation in the northwestern Iberian Peninsula. The aims were to assess the accumulation of herbivorous insects, including the distribution and frequency of herbivorous insects at different invasion levels (low, medium and high), and to evaluate whether introduced plants favour native or exotic, and generalist or specialist herbivorous insects in newly evolving plant–herbivore networks. To achieve these objectives, we surveyed A. dealbata along transects in five mixed pine forests and four shrublands, and C. edulis in nine coastal areas using quadrats. We identified nine herbivore species feeding on one or both of these species and determined that herbivore species composition in forest and coastal areas is directly tied to host preference, as well as related to invasion levels of A. dealbata in forest areas. On average, the introduced species in both the forest and coastal areas had approximately threefold more herbivore species (~ 50% of them exotics) than native plants. The introduced plants in forest areas had approximately eightfold more species interaction strength between herbivores and plants (a measure of the importance of each plant species to each herbivore). Overall, generalist herbivores were favoured by the invasion of the introduced plants. The results also demonstrate that exotic insects are supported by introduced plants, increasing the local occurrence and range of insect pests. Considering our observations, management strategies should be implemented to favour the restoration and protection of native areas for conserving biodiversity.

The Cuticle Mutant eca2 Modifies Plant Defense Responses to Biotrophic and Necrotrophic Pathogens and Herbivory Insects.

We isolated previously several Arabidopsis thaliana mutants with constitutive expression of the early microbe-associated molecular pattern-induced gene ATL2, named eca (expresión constitutiva de ATL2). Here, we further explored the interaction of eca mutants with pest and pathogens. Of all eca mutants, eca2 was more resistant to a fungal pathogen (Botrytis cinerea) and a bacterial pathogen (Pseudomonas syringae) as well as to a generalist herbivorous insect (Spodoptera littoralis). Permeability of the cuticle is increased in eca2; chemical characterization shows that eca2 has a significant reduction of both cuticular wax and cutin. Additionally, we determined that eca2 did not display a similar compensatory transcriptional response, compared with a previously characterized cuticular mutant, and that resistance to B. cinerea is mediated by the priming of the early and late induced defense responses, including salicylic acid- and jasmonic acid-induced genes. These results suggest that ECA2-dependent responses are involved in the nonhost defense mechanism against biotrophic and necrotrophic pathogens and against a generalist insect by modulation and priming of innate immunity and late defense responses. Making eca2 an interesting model to characterize the molecular basis for plant defenses against different biotic interactions and to study the initial events that take place in the cuticle surface of the aerial organs.

Why so many polyphagous fruit flies (Diptera: Tephritidae)? A further contribution to the ‘generalism’ debate

It has been suggested that generalist herbivorous insects are an evolutionary ‘improbability’. However, many species of tephritid fruit flies are generalists utilising larval hosts across two or more plant families. Focusing on the genus Bactrocera, it is argued that this high frequency of generalism is due to four evolutionary drivers which both decrease the need for specialism and promote generalism. Decreasing the need for specialism, there is firstly no co-evolutionary arms race between host plant and herbivore as Bactrocera do not negatively impact on plant fitness; this removes the need for specialism to counter plant defences. Secondly, the endemic hosts of most Bactrocera are plants with vertebrate-dispersed fleshy fruits. Fruit morphology and their display are highly constrained to attract those seed dispersers; Bactrocera thus do not need to specialize to increase host location capacity as many hosts produce a common signal. Thirdly, in native rainforest ecosystems Bactrocera are rare and so specialization to avoid competition seems unlikely. Promoting generalism are braconid parasitoids, which utilise plant cues to find their hosts. If a fly can use a novel host plant then it moves into enemy-free space, so promoting generalism. From this multi-part hypothesis, a group of testable predictions emerge. If shown to be correct then for the generalism/specialism debate frugivorous tephritids would provide an example where generalism may be a ‘probable’ evolutionary outcome.

Infestation of Caliothrips phaseoli (Thysanoptera: Thripidae) on Bean Cultivars Grown in the Winter, Rainy, and Dry Seasons in Brazil.

The present study aimed to identify common bean (Phaseolus vulgaris L.) cultivars less susceptible to Caliothrips phaseoli (Hood) in different growing seasons, to evaluate whether climatic conditions influence plant resistance to C. phaseoli infestation, and to investigate the preferred plant part for insect feeding. Eighteen common bean cultivars were evaluated in the winter season, and 19 cultivars were assessed in the rainy and dry seasons, under field conditions in the municipality of Jaboticabal, state of São Paulo, Brazil. Infestation of C. phaseoli nymphs in the upper and lower parts of the beans plants was recorded at weekly intervals from 25 days after plant emergence (DAE) to 60 DAE. In the winter season, the cultivars 'IAC Galante,' 'IAC Centauro,' 'IAC Carioca Eté,' and 'IAC Formoso' had significantly lower number of thrips than the cultivar 'IAC Diplomata.' In the rainy season, the cultivars 'IAC Harmonia' and 'IPR Siriri' had the lowest thrips infestation, differing from the cultivars 'BRS Pontal' and 'IAC Una.' The bean cultivars were equally susceptible to C. phaseoli in the dry season. The results suggest that C. phaseoli nymphs prefer to infest leaves of the lower part of bean plants, like most generalist herbivorous insects. In the winter and dry seasons, the highest thrips infestation was observed at 60 DAE, while in the rainy season, it was recorded from 32 to 46 DAE. Overall, C. phaseoli infestation on bean cultivars was not influenced by either temperature, relative humidity, or rainfall.

Landscape effects of a non-native grass facilitate source populations of a native generalist bug, Stenotus rubrovittatus, in a heterogeneous agricultural landscape.

Non-native plant species can provide native generalist insects, including pests, with novel food and habitats. It is hypothesized that local and landscape-level abundances of non-native plants can affect the population size of generalist insects, although generalists are assumed to be less sensitive to habitat connectivity than specialists. In a heterogeneous landscape in Japan, the relationship between the density of a native pest of rice (Stenotus rubrovittatus (Matsumura) (Heteroptera: Miridae)) and the abundance of Italian ryegrass (Lolium multiflorum Lam. (Poales: Poaceae)), a non-native meadow grass known to facilitate S. rubrovittatus, was analyzed. Statistical analyses of data on bug density, vegetation, and the spatial distribution of fallow fields and meadows dominated by Italian ryegrass, obtained by field surveys, demonstrated that local and landscape-level abundances of Italian ryegrass (the unmowed meadow areas within a few hundred meters of a sampling plot) positively affected bug density before its immigration into rice fields. Our findings suggest that a generalist herbivorous insect that prefers non-native plants responds to spatial availability and connectivity of plant species patches at the metapopulation level. Fragmentation by selective mowing that decreases the total area of source populations and increases the isolation among them would be an effective and environmentally-friendly pest management method.

Pollen mixing in pollen generalist solitary bees: a possible strategy to complement or mitigate unfavourable pollen properties?

Generalist herbivorous insects, which feed on plant tissue that is nutritionally heterogeneous or varies in its content of secondary metabolites, often benefit from dietary mixing through more balanced nutrient intake or reduced exposure to harmful secondary metabolites. Pollen is similarly heterogeneous as other plant tissue in its content of primary and secondary metabolites, suggesting that providing their offspring with mixed pollen diets might be a promising strategy for pollen generalist bees to complement nutrient imbalances or to mitigate harmful secondary metabolites of unfavourable pollen. In the present study, we compared larval performance of the pollen generalist solitary bee species Osmia cornuta (Megachilidae) on five experimental pollen diets that consisted of different proportions of unfavourable pollen diet of Ranunculus acris (Ranunculaceae) and favourable pollen diet of Sinapis arvensis (Brassicaceae). In addition, we microscopically analysed the pollen contained in the scopal brushes of field-collected females of O.cornuta and three closely related species to elucidate to what degree these pollen generalist bees mix pollen of different hosts in their brood cells. In striking contrast to a pure Ranunculus pollen diet, which had a lethal effect on most developing larvae of O.cornuta, larval survival, larval development time and adult body mass of both males and females remained nearly unaffected by the admixture of up to 50% of Ranunculus pollen diet to the larval food. Between 42% and 66% of all female scopal pollen loads analysed contained mixtures of pollen from two to six plant families, indicating that pollen mixing is a common behaviour in O.cornuta and the three related bee species. The present study provides the first evidence that the larvae of pollen generalist bees can benefit from the nutrient content of unfavourable pollen without being negatively affected by its unfavourable chemical properties if such pollen is mixed with favourable pollen. We conclude that the widespread pollen mixing by females of pollen generalist bees should also be considered as a possible strategy to exploit flowers with unfavourable pollen and to optimize larval food quality.

Tolerance for nutrient imbalance in an intermittently feeding herbivorous cricket, the Wellington tree weta.

Organisms that regulate nutrient intake have an advantage over those that do not, given that the nutrient composition of any one resource rarely matches optimal nutrient requirements. We used nutritional geometry to model protein and carbohydrate intake and identify an intake target for a sexually dimorphic species, the Wellington tree weta (Hemideina crassidens). Despite pronounced sexual dimorphism in this large generalist herbivorous insect, intake targets did not differ by sex. In a series of laboratory experiments, we then investigated whether tree weta demonstrate compensatory responses for enforced periods of imbalanced nutrient intake. Weta pre-fed high or low carbohydrate: protein diets showed large variation in compensatory nutrient intake over short (<48 h) time periods when provided with a choice. Individuals did not strongly defend nutrient targets, although there was some evidence for weak regulation. Many weta tended to select high and low protein foods in a ratio similar to their previously identified nutrient optimum. These results suggest that weta have a wide tolerance to nutritional imbalance, and that the time scale of weta nutrient balancing could lie outside of the short time span tested here. A wide tolerance to imbalance is consistent with the intermittent feeding displayed in the wild by weta and may be important in understanding weta foraging patterns in New Zealand forests.

Direct Proof of Ingested Food Regurgitation by Spodoptera littoralis Caterpillars during Feeding on Arabidopsis

Oral secretions of herbivorous lepidopteran larvae contain a mixture of saliva and regurgitant from the insect gut. Different compounds from the oral secretions can be recognized by the host plants and, thus, represent elicitors that induce plant defenses against feeding herbivores. Exogenously applied oral secretions can initiate the biosynthesis of jasmonates, phytohormones involved in the regulation of plant defense. However, it is not known (a) whether or not non-manipulated insects indeed release oral secretions including gut-derived compounds into a leaf wound during the natural feeding process, or (b) whether they adjust the release of gut components to the state of plant defense. We addressed these questions by using Arabidopsis thaliana as host plant and larvae of the generalist herbivorous insect Spodoptera littoralis. We investigated the conversion of the plant-derived jasmonate precursor, cis-12-oxophytodienoic acid (cis-OPDA), to iso-OPDA by the larvae. This enzymatic reaction is mediated by a specific glutathione-S-transferase in the insect gut, but not in the plant. Any presence of iso-OPDA in plant tissue, thus, indicated that gut content had been regurgitated into the plant wound. Our study demonstrates that the plant is the only source for the substrate cis-OPDA by using aos (allene oxide synthase) mutants that are unable to synthesize OPDA. The fact that iso-OPDA accumulated over time on feeding-damaged leaves shows that the feeding larvae are constantly regurgitating on leaves. Although the larvae provided the signaling compounds that were recognized by the plant and elicited defense reactions, the larval regurgitation behavior did not depend on whether they fed on a defensive wild type plant or on a non defensive coi1-16 plant. This suggests that S. littoralis larvae do not adjust regurgitation to the state of plant defense.

CML42-mediated calcium signaling coordinates responses to Spodoptera herbivory and abiotic stresses in Arabidopsis.

In the interaction between Arabidopsis (Arabidopsis thaliana) and the generalist herbivorous insect Spodoptera littoralis, little is known about early events in defense signaling and their link to downstream phytohormone pathways. S. littoralis oral secretions induced both Ca²⁺ and phytohormone elevation in Arabidopsis. Plant gene expression induced by oral secretions revealed up-regulation of a gene encoding a calmodulin-like protein, CML42. Functional analysis of cml42 plants revealed more resistance to herbivory than in the wild type, because caterpillars gain less weight on the mutant, indicating that CML42 negatively regulates plant defense; cml42 also showed increased aliphatic glucosinolate content and hyperactivated transcript accumulation of the jasmonic acid (JA)-responsive genes VSP2 and Thi2.1 upon herbivory, which might contribute to increased resistance. CML42 up-regulation is negatively regulated by the jasmonate receptor Coronatine Insensitive1 (COI1), as loss of functional COI1 resulted in prolonged CML42 activation. CML42 thus acts as a negative regulator of plant defense by decreasing COI1-mediated JA sensitivity and the expression of JA-responsive genes and is independent of herbivory-induced JA biosynthesis. JA-induced Ca²⁺ elevation and root growth inhibition were more sensitive in cml42, also indicating higher JA perception. Our results indicate that CML42 acts as a crucial signaling component connecting Ca²⁺ and JA signaling. CML42 is localized to cytosol and nucleus. CML42 is also involved in abiotic stress responses, as kaempferol glycosides were down-regulated in cml42, and impaired in ultraviolet B resistance. Under drought stress, the level of abscisic acid accumulation was higher in cml42 plants. Thus, CML42 might serve as a Ca²⁺ sensor having multiple functions in insect herbivory defense and abiotic stress responses.

Chemical characterization of the aphid antifeedant extracts from Dittrichia viscosa and Ferula communis

Abstract The plant defensive activities of the hexane and ethanolic extracts from Dittrichia viscosa and Ferula communis against generalist herbivorous insects ( Spodoptera littoralis and Myzus persicae ), are described here. Additionally we tested for their effects on nematodes ( Meloidogyne javanica ) and plants ( Lactuca sativa and Lolium perenne ). These plant extracts showed variable biological effects, their aphid antifeedant action being the most important. The bioassay-guided search for aphid antifeedant compounds resulted in the isolation of tomentosin ( 1 ), inuviscolide ( 2 ) and ferulenol ( 3 ). Tomentosin ( 1 ) rich extracts of D. viscosa were more active than ferulenol ( 3 ) rich extracts from F. communis.

Silencing Defense Pathways in Arabidopsis by Heterologous Gene Sequences from Brassica oleracea Enhances the Performance of a Specialist and a Generalist Herbivorous Insect

The jasmonic acid (JA) signaling pathway and defensive secondary metabolites such as glucosinolates are generally considered to play central roles in the defense of brassicaceous plants against herbivorous insects. To determine the function of specific plant genes in plant-insect interactions, signaling or biosynthetic mutants are needed. However, mutants are not yet available for brassicaceous plants other than Arabidopsis thaliana, e.g., cabbage (Brassica oleracea). We employed virus-induced gene silencing (VIGS) by using tobacco rattle virus (TRV) to knock down the endogenous expression of lipoxygenase (LOX), an upstream enzyme of the JA pathway and thioglucoside glucohydrolase: myrosinase (TGG1/TGG2), a hydrolytic enzyme that catalyzes the release of defensive volatile products originating from glucosinolates, in Arabidopsis thaliana. This was done by using the heterologous gene sequences from B. oleracea. Silencing these genes in A. thaliana plants is efficient and specific. Only 18 nucleotides with 100% identity between the trigger (BoMYR) and the target (AtTGG1/2) sequence are sufficient to achieve gene silencing. LOX-silenced plants showed significantly reduced AtLOX2 transcript accumulation after Pieris rapae larval feeding. TGG-silenced plants exhibited significantly lower TGG1/TGG2 transcript levels only after shorter larval feeding. The inhibition of TGG1/TGG2 transcript accumulation via gene silencing may be overruled by longer larval feeding. Specialist P. rapae larvae developed significantly better on both types of silenced plants than on empty vector (EV) control plants, while generalist Mamestra brassicae larvae developed significantly better on the TGG1/TGG2 silenced plants than on EV control plants. This shows that not only the generalist herbivore but also the Brassicaceae-specialist P. rapae is negatively affected by the ability of brassicaceous plants to produce their specific secondary metabolites, i.e., glucosinolates. Our results demonstrate the important roles of AtLOX2 and AtTGG1/TGG2 genes, which were silenced by heterologous gene sequences from B. oleracea BoLOX and BoMYR, in A. thaliana resistance to the specialist P. rapae and the generalist M. brassicae.

Mapping the Larval Midgut Lumen Proteome of Helicoverpa armigera, a Generalist Herbivorous Insect

The gut lumen is the primary site of digestion and detoxification and thus presents conditions hostile to most proteins. We used 2D-gel electrophoresis and MS/MS de novo peptide sequencing to identify the major proteins stable enough to persist in the midgut lumen of caterpillars of the cotton bollworm Helicoverpa armigera, a generalist herbivorous insect and a major crop pest worldwide. As expected, we found several enzymes responsible for digestion of carbohydrates, proteins, and lipids. In addition, we identified nondigestive proteins such as a multidomain lipocalin, a protein with pathogen recognition domains, an arginine kinase related to a class of major human allergens, and abundant proteins of unknown function. Identification of the set of proteins that are secreted into the lumen will enable us to further characterize the nutritional and defensive functions of this important intraorganismal space.

Effects of synthesized tannin on the growth and survival of a generalist herbivorous insect, the common cutworm, Spodoptera litura Fabricius (Lepidoptera: Noctuidae).

We examined the effects of tannin on a generalist herbivorous insect, the common cutworm (Spodoptera litura). The dosage effect was estimated by measuring the growth performance of the cutworm when the larvae were fed artificial diets containing different amounts of synthesized tannin. We used artificial diets to exclude the effects of non-tannin defense mechanisms in plant foliage. Indices of growth performance, number of dead larvae, pupal weight, total feeding amount and length of larval period were obtained. We found that tannin inhibited the growth of the cutworm larvae and that the inhibitory effect was proportional to the amount of tannin ingested. These results support the traditional theory that plant tannin is a defense agent against generalist herbivores, and that its effects are proportional to its concentration in foliage.

Seasonality of insect herbivory on the leaves of Neoboutonia macrocalyx in the Kibale National Park, Uganda

SummaryThe seasonality of herbivory on the leaves of Neoboutonia macrocalyx Pax. in Kibale Forest National Park, Uganda, was studied. A total of 2929 fallen leaves was collected during 15 months under randomly‐selected trees in three different habitats; natural forest and two selectively cut forest sites. The percentage of leaf area eaten and leaf size were estimated. Leaf herbivory was highly seasonal and correlated with rainfall in the previous 2 months, but less than 100 mm monthly rainfall had no effect. There was no correlation between leaf size and rainfall. Although Kibale Forest has two wet seasons, insect feeding on leaves had only one peak during the major rainy season from September to December. Three to four months after peak herbivory, leaves had very low rates of insect damage. Habitat had only a small effect on the amount of insect feeding. The sampling time accounted for 71% of variation in leaf herbivory. New leaves were formed continuously year‐round. The constant leaf production by Neoboutonia trees may be an adaptation to escape generalist herbivorous insects which might be synchronized with the major wet season when the leaf flush of the most other deciduous species occurs. Thus, the availability of fresh leaves is not acting as a regulating factor in seasonality of Neoboutonia herbivory.

Defensive role ofAllium sulfur compounds for leek mothAcrolepiopsis assectella Z. (Lepidoptera) against generalist predators

It has been shown previously that sulfur volatiles produced byAllium plants affect the behavior of their specialist phytophages and of their specialist entomophages. The action of these compounds in protecting the leek mothAcrolepiopsis assectella against generalist entomophages was studied in comparison to the proposed original defensive role of these compounds against generalist herbivorous insects. Two ants species,Formica selysi andF. fusca, were used as generalist predators. Six behavioral criteria of the predatory behavior of the ants were studied in presence of the last-instar caterpillars (C5). C5 reared on artificial diets with or without leek components were tested, as well as C5 soaked in frass of leek-reared caterpillars or disulfide solutions. In addition, the response of the ants to pure chemicals found in leek was studied using honey solutions with or without sulfur compounds. The sulfur allelochemicals ofAllium plants have a negative action on predatory ants. Interestingly, the nonvolatile precursors of sulfur volatiles ofAllium plants seem to have a protective role for their phytophagous insects against generalist entomophages.