Generalist Herbivores Research Articles

Precipitation Drives Frugivory in a Subtropical Generalist Herbivore, the Gopher Tortoise, and Alters Its Functional Role as a Seed Disperser.

Consumers employ a variety of foraging strategies, and oftentimes the foraging strategy employed is related to resource availability. As consumers acquire resources, they may interact with their resource base in mutualistic or antagonistic ways-falling along a mutualism-antagonism continuum-with implications for ecological processes such as seed dispersal. However, patterns of resource use vary temporally, and textbook herbivores may switch foraging tactics to become more frugivorous in periods of greater fleshy fruit availability. In this study, we investigated how fleshy fruit consumption of a generalist herbivore-the gopher tortoise (Gopherus polyphemus)-shifts intra-annually following seasonal precipitation and subsequently examined how this shift toward increased frugivory influences the suite of plant syndromes dispersed. We noted a clear intra-annual shift toward a more frugivorous diet which coincided with seasonal precipitation and subsequently observed a marked shift in the plant syndromes dispersed with increasing frugivory. We found that as this generalist herbivore became more frugivorous, it dispersed a greater variety of plant syndromes at low levels of frugivory. However, when the gopher tortoise exhibited high levels of frugivory, the seed load was dominated by those exhibiting the endozoochory syndrome. This study illustrates a functional shift in a seed-dispersing herbivore toward that of a classical frugivore, suggesting that temporal variation in foraging strategy and the temporal scale in which foraging habits and seed dispersal interactions are quantified have implications for the suite of plant syndromes species disperse. Furthermore, trade-offs may exist that provide plants with the endozoochory syndrome with a competitive advantage over seeds with contrasting traits, such as the foliage is the fruit syndrome which is expected to experience greater dispersal by classical herbivores.

The fate of a Solanum steroidal alkaloid toxin in the cabbage looper (Trichoplusia ni)

Plants produce complex chemical defenses against herbivores, resulting in the emergence of detoxification strategies in phytophagous insects. While enzymatic detoxification and target site mutagenesis are well-documented, the quantitative contribution of excretion remains less studied. We focus on the cabbage looper (Trichoplusia ni), a generalist herbivore, to elucidate the detoxification of a steroidal alkaloid, solanidine, produced in potato (Solanum tuberosum). Through larval feeding experiments and chemical analysis of metabolites using high-resolution mass spectrometry, we identify solanidine 3-O-β-glucopyranoside and solanidine 3-phosphate as major metabolization products of solanidine. Glycosylation and phosphorylation reactions have not previously been observed in cabbage looper. Modified solanidine derivatives exhibit reduced lipophilicity, preventing passive transport as predicted by physicochemical analyses, and only solanidine was detected in body tissue. In addition, the metabolism of solanidine in a T. ni mutant strain with midgut cadherin protein knocked out was also investigated to examine the potential role of the cadherin, an important receptor for Bt toxins, in steroidal alkaloid detoxification. T. ni cadherin-knockout strain showed lower solanidine conversion (33.9% ± 2.2) and uptake (27.41 ± 0.49 nmol/g) compared to the wild-type strain (51.3% ± 4.1, 33.66 ± 2.48 nmol/g) but similar excretion kinetics. Although solanidine negatively impacted the feeding performance of both strains the cadherin-knockout does not affect the feeding performance. Our study expands the metabolization enzyme repertoire in cabbage loopers, emphasizing the complexity of detoxification mechanisms in generalist herbivores.

Diet of the crested porcupine Hystrix cristata (Rodentia, Hystricidae) in a semi-arid area of its native North African range

AbstractWe investigated the feeding ecology of the crested porcupine Hystrix cristata in a semi-arid area of its northwestern Algerian native range over 8 months (September to April). Our results from micro-histological analysis based on faecal pellets revealed that this strict herbivore ate only 10 plant species, 8 wild and 2 cultivated, belonging to 9 families including Lamiaceae, Palmaceae, Poaceae, Ramnaceae and Fabaceae. The autumn diet was dominated by Thymus algeriensis, Ziziphus lotus, Chamaerops humilis and Triticum aestivum, the winter diet by T. algeriensis and C. humilis, and the spring diet by Muscari comosum, with a large amount of Vicia faba and T. aestivum. We confirm the role of this rodent species as a generalist herbivore which barely feeds on cultivated plants when wild resources are diverse and abundant.

Geographic variation in leaf traits and palatability of a native plant invader during domestic expansion.

Like alien plant invasion, range expansion of native plants may threaten biodiversity and economies, rendering them native invaders. Variation in abiotic and biotic conditions across a large geographic scale greatly affects variation in traits and interactions with herbivores of native plant invaders, which is an interesting yet mostly unexplored issue. We used a common garden experiment to compare defensive/nutritional traits and palatability to generalist herbivores of 20 native (23.64° N-30.18° N) and introduced range (31.58° N-36.87° N) populations of Reynoutria japonica, which is a native invader following range expansion in China. We analyzed the relationships among herbivore pressure, climate, plant chloroplast haplotypes, leaf traits, and herbivore performance. Of the 16 variables tested, we observed range differences in 11 variables and latitudinal clines in nine variables. In general, herbivores performed better on the introduced plants than on the native plants, and better on the high-latitude plants than on the low-latitude plants within the introduced populations. Three key traits (leaf thickness, specific leaf area, and carbon-to-nitrogen [C:N] ratio) determined palatability to herbivores and were significantly associated with temperature and/or precipitation of plant provenance as well as with plant haplotypes but not with herbivore pressure. Our results revealed a causal sequence from plant-range-based environmental forces and genetic context to plant quality and palatability to herbivores in R. japonica. These findings suggest a post-introduction evolution of R. japonica, which may partly explain the colonization success of this important native, but invasive plant.

Decoding plant-induced transcriptomic variability and consistency in two related polyphagous mites differing in host ranges.

The diet breadth of generalist herbivores when compared to specialists tends to be associated with greater transcriptional plasticity. Here, we consider whether it may also contribute to variation in host range among two generalists with different levels of polyphagy. We examined two related polyphagous spider mites with different host ranges, Tetranychus urticae (1200 plants) and Tetranychus truncatus (90 plants). Data from multiple populations of both species domesticated on common beans and transferred to new plant hosts (cotton, cucumber, eggplant) were used to investigate transcriptional plasticity relative to population-based variation in gene expression. Compared to T. truncatus, T. urticae exhibited much higher transcriptional plasticity. Populations of this species also showed much more variable expression regulation in response to a plant host, particularly for genes related to detoxification, transport, and transcriptional factors. In response to the different plant hosts, both polyphagous species showed enriched processes of drug/xenobiotics metabolism, with T. urticae orchestrating a relatively broader array of biological pathways. Through co-expression network analysis, we identified gene modules associated with host plant response, revealing shared hub genes primarily involved in detoxification metabolism when both mites fed on the same plants. After silencing a shared hub CYP gene related to eggplant exposure, the performance of both species on the original bean host improved, but the fecundity of T. truncatus decreased when feeding on eggplant. The extensive transcriptomic variation shown by T. urticae might serve as a potential compensatory mechanism for a deficiency of hub genes in this species. This research points to nuanced differences in transcriptomic variability between generalist herbivores.

Short-term responses of spider mites inform mechanisms of maize resistance to a generalist herbivore

Plants are attacked by diverse herbivorous pests with different host specializations. While host plant resistance influences pest pressure, how resistance impacts the behaviors of generalist and specialist herbivores, and the relationship to resistance, is less well known. Here, we investigated the short-term (< 1 h) behavioral changes of a generalist herbivore, the two-spotted spider mite (TSM), and a specialist herbivore, the Banks grass mite (BGM), after introduction to no-choice Tanglefoot leaf-arenas (2 × 2 cm) of three maize inbred lines (B73, B75, and B96). The widely-used inbred line B73 is susceptible to spider mites, while B75 and B96 are known to be mite resistant, especially to TSM. Video tracking was used to record TSM and BGM walking, probing, feeding, resting, web-building and travel distance on arenas of each line. Mite oviposition was also recorded after 72 h. B75, a resistant line, decreased the feeding behavior (i.e., time) of both mite species compared to B73 (susceptible control) and B96. Moreover, TSM appeared to be sensitive to both resistant lines (B75 and B96) with reduced oviposition, and increased resting and web-building times compared to susceptible B73. In contrast, the specialist BGM showed no difference in oviposition, resting and web-building time across all maize inbred lines. Our findings of quite broad and short-term responses of TSM to B75 and B96 are consistent with a role for constitutive or rapidly induced plant defenses in maize in conferring TSM resistance. Other mechanisms of plant resistance may be needed, however, for defense against specialists like BGM.

Variation in immune response in the generalist herbivore fall webworm across four common host plants

AbstractDietary generalist herbivorous insects are widespread and often occur in a variety of environments. Across their geographic range, herbivorous insects may encounter variable plant traits as they feed on high‐quality or low‐quality plants. Herbivorous insect larvae experience both bottom‐up (host plant) and top‐down (parasitoid) factors that affect survival. Host plant quality may affect larval growth and survival in that larvae feeding on low‐quality plants often suffer reduced fitness. However, herbivores on different host plants are also subject to different levels of parasitism. High‐quality plants confer stronger larval performance (higher survival, more offspring), but larvae may also face higher parasitism. In some herbivore species, diet mediates larval immune response. The generalist insect herbivore fall webworm (FW), Hyphantria cunea Drury (Lepidoptera: Erebidae), is a moth native to North America, and its larvae have considerable variance in their performance when reared on different host plants. We investigated whether diet affects the immune response in FW larvae when they are reared on different host plant species known to vary in food quality. We measured immune response by melanization of a nylon filament. We found significant differences in immune response across host plants, indicating that diet mediates immune response in FW larvae. Our study helps elucidate the factors that cause variation in immune response in a generalist herbivore.

Changes in tomato plant anti-herbivore defences after soil application of a biofungicide containing Bacillus subtilis (Serenade ASO)

ABSTRACT Plants have limited resources to allocate to defences against infection and herbivory. While interactions between plant responses to microbial and herbivore attack are complex, it is often the case that the induction of one response will act antagonistically to the other. Recent studies have shown that plant growth promoting rhizobacteria, which improves overall plant health and general stress resistance, can enhance both anti-microbial and anti-herbivore defences. We investigated how soil application of the biofungicide Serenade ASO (Bacillus subtilis strain QST 713), which primes plant defences against fungal and bacterial infection and promotes plant growth, affects anti-herbivore defences by measuring both constitutive and induced defences. We applied Serenade one or two times to the soil of tomato plants and measured the numbers of type IV glandular trichomes on leaves, the weight gain of a generalist caterpillar (beet armyworms; BAW), and the activity of two enzymes associated with defence against insects (polyphenoloxidase and peroxidase). Serenade treated plants grew faster and foliage from treated plants had significantly higher numbers of glandular trichomes and higher polyphenoloxidase and peroxidase activities than untreated plants. However, Serenade treatment did not affect the degree of induction of plant defences when damaged by BAW feeding and did not slow the growth rate of BAW relative to plants not treated with Serenade. Therefore, the biofungicide Serenade increased plant growth and altered the densities of trichomes and the activities of two defensive enzymes in plants, but it did not affect overall susceptibility of the plants to a generalist herbivore.

Assessing grazing of Rugulopteryx okamurae (Ochrophyta) by the sea urchin Paracentrotus lividus (Echinodermata): a prospect for the biocontrol of the alien species?

ABSTRACT Rugulopteryx okamurae is an invasive non-indigenous species (NIS) of marine alga present in the benthic communities of the Strait of Gibraltar, where it has colonized most of the coast triggering ecological, economic and social consequences. The purple sea urchin Paracentrotus lividus is a generalist herbivore with potential to provide resistance to this NIS. Two experiments were carried out to determine if (i) P. lividus consumes R. okamurae under laboratory conditions, (ii) to assess its grazing preferences when this NIS is offered together with the native Ulva sp. and (iii) to find biochemical tissue markers related to R. okamurae ingestion. In the first experiment, sea urchins fed with R. okamurae, Ulva sp. or both diets were evaluated in groups for two months. Ingestion, food preference, growth and survival were assessed. In the second one, the same specimens from the R. okamurae and mixed diets were used to analyse their individual preferences, ingestion rates, growth and fatty acid profiles during another two months. Ingestion rates showed that sea urchins accepted the NIS R. okamurae as food under laboratory conditions, increasing its intake after a long exposure period. The electivity values for R. okamurae also increased when P. lividus was exposed to this alga for a long time. Regarding the fatty acids profile of tissues, the results showed that intestinal arachidonic acid (ARA, 20:4n-6) clearly differentiates sea urchins fed with R. okamurae from those fed with the mixed diet. ARA also appeared to be related to the feeding preferences, highlighting this fatty acid as a reliable marker of R. okamurae ingestion. Further research is needed in order to determine P. lividus feeding behaviour and its role as a biocontrol species in the areas invaded by R. okamurae.

Patterns of constitutive and induced herbivore defence are complex, but share a common genetic basis in annual and perennial monkeyflower

Abstract Despite multiple ecological and evolutionary hypotheses that predict patterns of phenotypic relationships between plant growth, reproduction and constitutive and/or induced resistance to herbivores, these hypotheses do not make any predictions about the underlying molecular genetic mechanisms that mediate these relationships. We investigated how divergent plant life‐history strategies in the yellow monkeyflower and a life‐history altering locus, DIV1, influence plasticity of phytochemical herbivory resistance traits in response to attack by two herbivore species with different diet breadth. Life‐history strategy (annual vs. perennial) and the DIV1 locus significantly influenced levels of constitutive herbivory resistance, as well as resistance induction following both generalist and specialist herbivory. Perennial plants had higher total levels of univariate constitutive and induced defence than annuals, regardless of herbivore type. Annuals induced less in response to generalist herbivory than did perennials, while induction response was equivalent across the ecotypes for specialist herbivory. The effects of the DIV1 locus on levels of constitutive and induced defence were dependent on genetic background, the annual versus perennial haplotype of DIV1 and herbivore identity. The patterns of univariate induction due to DIV1 were non‐additive and did not always match expectations based on patterns of divergence for annual/perennial parents. For example, perennial plants had higher levels of constitutive and induced defence than did annuals, but when the annual DIV1 was present in the perennial genetic background induction response to herbivory was higher than for the perennial parent lines. Patterns for multivariate defence arsenals generally echoed those of univariate, with annual and perennial monkeyflowers and those with alternative versions of DIV1 differing significantly in constitutive and induced resistance. Like univariate resistance, induced multivariate defence arsenals were affected by herbivore identity. Our results highlight the complexity of the genetic mechanisms underlying plastic response to herbivory. While a genetic locus underlying substantial phenotypic variation in life‐history strategy and constitutive defence also influences defence plasticity, the induction response also depends on genetic background. This result demonstrates the potential for some degree of evolutionary independence between constitutive and induced defence, or induced defence and life‐history strategy, in monkeyflowers. Read the free Plain Language Summary for this article on the Journal blog.

Developing Oak Buds Produce Volatile Emissions in Response to Herbivory by Freshly Hatched Caterpillars.

Plant responses to damage by insectivorous herbivores are well-documented in mature leaves. The resulting herbivore-induced plant volatiles (HIPVs) protect the plant by attracting carnivorous arthropods and even some insectivorous vertebrates, to parasitize or consume the plant invaders. However, very little is known about plant production of HIPVs in developing buds, particularly when herbivorous insects are too small to be considered a prey item. It is additionally unclear whether plants respond differently to generalist and specialist chewing insects that overlap in distribution. Therefore, we compared HIPV production of Downy oak (Quercus pubescens Willd.) buds infested with freshly hatched caterpillars of Tortrix viridana (specialist) and Operophtera brumata (generalist), against uninfested buds. Of the compounds identified in both years of the experiment, we found that (Z)-hex-3-enyl acetate, (E)-β-ocimene, acetophenone, linalool, (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), methyl salicylate, α-copaene, α-humulene, (E)-caryophyllene, and (E,E)-α-farnesene appeared to be higher in infested buds compared to controls. We found no difference in HIPV production between the specialist and the generalist herbivores. Production of HIPVs was also associated with leaf damage, with higher HIPV production in more severely attacked buds. Thus, our study shows that oak trees already start responding to insect herbivory before leaves are developed, by producing compounds similar to those found in damaged mature leaves. Future work should focus on how Downy oak may benefit from initiating alarm cues at a time when carnivorous arthropods and insectivorous vertebrates are unable to use herbivorous insects as host or food.

The transition from diet to blood: Exploring homeostasis in the insect haemolymph nutrient pool

AbstractNutrition is vital to health, but while the link between diet and body nutritional composition is well explored in humans and other vertebrates, this information is not well understood in insects, despite the vital roles they play in ecosystems, and their increasing use as experimental models. Here we used Nutritional Geometry to explore the rapid physiological response to ingested nutrients in the haemolymph nutritional profile of Spodoptera littoralis caterpillars. We ask whether blood nutrients are maintained homeostatically in the face of variable nutritional intake, or if regulation is more flexible for some nutrients than others (allostasis), which allows animals to adapt to stress by responding in a way that prioritises efficiency of responses in the face of trade‐offs. Caterpillars were placed on 1 of 20 diets, systematically varying in their nutrient ratios (protein: carbohydrate) and density (calorie content), and their consumption was measured. After 48 h, caterpillars were bled, and the macronutrient (protein, carbohydrates and lipids) and nutrient metabolite (amino acids and simple sugars) content of the haemolymph was measured. Proteins comprised 93% of the haemolymph macronutrient pool on average and their concentration increased with protein eaten. The amino acid (AA) pool was dominated by five AAs, and the total pool increased with total nutrient intake. However, the ratio of essential to non‐essential AAs increased as the proportion of protein consumed increased. Carbohydrates were tightly controlled, increasing only on the most extreme carbohydrate intakes. Simple sugars were dominated by glucose and trehalose, and overall, the simple sugar pool showed high levels of homeostasis. Rather than strict homeostasis of blood nutritional properties, an allostatic model seemed to be a better fit for blood nutrient regulation in this generalist herbivore. This flexibility in response to the nutritional composition of the diet may, in part, explain how this species has evolved to extreme dietary generalism and may play a role in its worldwide pest status. Given the range of fitness‐related processes affected by the haemolymph, future studies should examine the physiological impacts of blood nutrient variation on reproduction, growth and response to infection and the trade‐offs between them.

Gastropod grazing on fresh and senescent leaves of non-native invasive plants Reynoutria japonica and Impatiensglandulifera

In invaded regions, the introduced non-native plants Reynoutria japonica and Impatiens glandulifera show little grazing damage, most likely due to the absence of specialized herbivores and the deterrent effect of secondary metabolites on generalist herbivores. The low degree of grazing damage has been explained by the non-mutually exclusive enemy-release and novel weapon hypotheses. We tested assumptions of these hypotheses by conducting a series of preference tests in which leaf samples from R. japonica, I. glandulifera, and the native Urtica dioica were offered to five species of generalist gastropods (the native Arianta arbustorum, Cepaea nemoralis and Fruticicola fruticum, and the non-native invasive Hygromia cinctella and Arion vulgaris). In addition, we determined the C/N-ratio and total phenolic compounds (as a surrogate of secondary metabolites) of the plant species. In the choice experiment with fresh leaf samples, all snail species showed a preference for U. dioica, with the exception of the non-native invasive slug A. vulgaris, which ate leaf tissue from I. gladulifera almost as much as from U. dioica. The snails’ preference of U. dioica was even more pronounced when the fresh weight of leaf material eaten was considered. No-choice tests with either fresh or dead R. japonica leaves showed that most individuals of all species ate small amounts of fresh leaves, but less of dead leaves. In contrast, no-choice tests with either fresh or senescent I. glandulifera leaf tissue showed that individuals of all five gastropod species consumed larger amounts of senescent leaves than fresh leaves, probably because secondary compounds have been broken down or leached or the leaf texture has changed. The low susceptibility of these non-native invasive plants to the gastropods can most likely be explained by a combination of chemical and physical plant characteristics.

Struggling to survive: A comparison of Vanessa cardui larval survivorship on putative host plants

AbstractThe painted lady butterfly (Vanessa cardui) is a generalist herbivore with a global distribution. In North America, over 100 species have been identified as V. cardui larval host plants. This cosmopolitan species is commercially available in all life stages and can be reared on an artificial diet. As a result, V. cardui commonly serves as a model organism for education and outreach. However, gaps in our knowledge remain with regard to V. cardui ecology and larval host plant suitability for supporting complete development of larvae to reproductive adulthood. In a laboratory setting, we tested host plant suitability of five reported host plants commonly found in California sage scrub ecosystems by assessing survival of V. cardui caterpillars in its entirety. We also assessed how commercially sourced larvae survived on artificial diet: both raising the caterpillars exclusively on artificial diet as well as switching diets from artificial to thistle (a commonly reported host plant) and switching from thistle to artificial diet. We found that all commercially sourced caterpillars exhibited both high larval mortality and low pupation rates when reared on host plants, although larvae reared on Malacothamnus fasciculatus and Sphaeralcea ambigua survived longest. Moreover, M. fasciculatus and S. ambigua were the only host plants tested that supported successful pupation. This contrasted with 63% higher survival of wild‐collected larvae. Our findings suggest that commercially obtained V. cardui may struggle to utilize wild host plants and that future investigations into host plant suitability in wild populations are needed. Additionally, future research using commercial larvae should consider the implications for interpreting host plant suitability as larvae may exhibit adaptations to artificial diet or the loss of adaptation to consuming plant material.

Food and time: dietary plasticity of different sources of a generalist insect herbivore.

Painted lady butterflies (Vanessa cardui L.,Nymphalidae) are generalist herbivores and serve as a model system across several fields of biology. While it has been demonstrated that V. cardui caterpillars can develop on different host plants, much of this work has been done on commercially sourced caterpillars, which could limit our understanding of wild V. cardui populations. In this study, we sought to explore possible differences in how commercial and wild V. cardui caterpillars may respond to feeding on different host plants, and subsequently, how their diet impacts immune response and survival. Here, we analyzed performance, survival, and immune response of wild and commercially sourced V. cardui caterpillars over several generations on diets that consisted of either 1 of 4 different host plant species or a mixed diet including all 4 species. Qualitatively, we observed that wild larvae had a better larval performance and hemocyte counts compared to the commercial larvae. The results demonstrate that both wild and commercially sourced caterpillars grew and survived best on the same diet treatments (mallow, narrowleaf plantain, and a mixed diet) during development across generations. Immune responses showed similar patterns across host plants between wild and commercial populations, with individuals showing lowered immune responses on dandelion and lupine and higher ones on mallow, plantain and the mixed diet; although the relative rankings on those 3 diets varied. Survival also demonstrated similar patterns, in that individuals reared on dandelion and lupine had the lowest survival.

Aphid Resistance Segregates Independently of Cardenolide and Glucosinolate Content in an Erysimum cheiranthoides (Wormseed Wallflower) F2 Population.

Plants in the genus Erysimum produce both glucosinolates and cardenolides as a defense mechanism against herbivory. Two natural isolates of Erysimum cheiranthoides (wormseed wallflower) differed in their glucosinolate content, cardenolide content, and their resistance to Myzus persicae (green peach aphid), a broad generalist herbivore. Both classes of defensive metabolites were produced constitutively and were not further induced by aphid feeding. To investigate the relative importance of glucosinolates and cardenolides in E. cheiranthoides defense, we generated an improved genome assembly, genetic map, and segregating F2 population. The genotypic and phenotypic analysis of the F2 plants identified quantitative trait loci, which affected glucosinolates and cardenolides, but not the aphid resistance. The abundance of most glucosinolates and cardenolides was positively correlated in the F2 population, indicating that similar processes regulate their biosynthesis and accumulation. Aphid reproduction was positively correlated with glucosinolate content. Although the overall cardenolide content had little effect on aphid growth and survival, there was a negative correlation between aphid reproduction and helveticoside abundance. However, this variation in defensive metabolites could not explain the differences in aphid growth on the two parental lines, suggesting that processes other than the abundance of glucosinolates and cardenolides have a predominant effect on aphid resistance in E. cheiranthoides.

Specialist reassociation and residence time modulate the evolution of defense in invasive plants: A meta-analysis.

Invasive plants typically escape specialist herbivores but are often attacked by generalist herbivores in their introduced ranges. The shifting defense hypothesis suggests that this will cause invasive plants to evolve lower resistance against specialists, higher resistance against generalists, and greater tolerance to herbivore damage. However, the duration and direction of selective pressures can shape the evolutionary responses of resistance and tolerance for invasive plants. Two critical factors are (1) residence time (length of time that an invasive species has been in its introduced range) and (2) specialist herbivore reassociation (attack by purposely or accidentally introduced specialists). Yet, these two factors have not been considered simultaneously in previous quantitative syntheses. Here, we performed a meta-analysis with 367 effect sizes from 70 studies of 35 invasive plant species from native and invasive populations. We tested how the residence time of invasive plant species and specialist reassociation in their introduced ranges affected evolutionary responses of defenses against specialists and generalists, including herbivore resistance traits (physical barriers, digestibility reducers and toxins), resistance effects (performance of and damage caused by specialists or generalists) and tolerance to damage (from specialists or generalists). We found that residence time and specialist reassociation each significantly altered digestibility reducers, specialist performance, generalist damage, and tolerance to specialist damage. Furthermore, residence time and specialist reassociation strongly altered toxins and generalist performance, respectively. When we restricted consideration to invasive plant species with both longer residence times and no reassociation with specialists, invasive populations had lower resistance to specialists, similar resistance to generalists, and higher tolerance to damage from both herbivore types, compared with native populations. We conclude that the duration and direction of selective pressure shape the evolutionary responses of invasive plants. Under long-term (long residence time) and stable (no specialist reassociation) selective pressure, invasive plants generally decrease resistance to specialists and increase tolerance to generalist damage that provides mixed support for the shifting defense hypothesis.

Cardiac glycosides protect wormseed wallflower (Erysimum cheiranthoides) against some, but not all, glucosinolate-adapted herbivores.

The chemical arms race between plants and insects is foundational to the generation and maintenance of biological diversity. We asked how the evolution of a novel defensive compound in an already well-defended plant lineage impacts interactions with diverse herbivores. Erysimum cheiranthoides (Brassicaceae), which produces both ancestral glucosinolates and novel cardiac glycosides, served as a model. We analyzed gene expression to identify cardiac glycoside biosynthetic enzymes in E. cheiranthoides and characterized these enzymes via heterologous expression and CRISPR/Cas9 knockout. Using E. cheiranthoides cardiac glycoside-deficient lines, we conducted insect experiments in both the laboratory and field. EcCYP87A126 initiates cardiac glycoside biosynthesis via sterol side-chain cleavage, and EcCYP716A418 has a role in cardiac glycoside hydroxylation. In EcCYP87A126 knockout lines, cardiac glycoside production was eliminated. Laboratory experiments with these lines revealed that cardiac glycosides were highly effective defenses against two species of glucosinolate-tolerant specialist herbivores, but did not protect against all crucifer-feeding specialist herbivores in the field. Cardiac glycosides had lesser to no effect on two broad generalist herbivores. These results begin elucidation of the E. cheiranthoides cardiac glycoside biosynthetic pathway and demonstrate in vivo that cardiac glycoside production allows Erysimum to escape from some, but not all, specialist herbivores.

Investigating the effect of host plant identity on instar number in fall webworm, a common generalist herbivore.

For herbivorous insects with a broad diet breadth, host plant identity can influence larval development by either accelerating or delaying growth. For some species of Lepidoptera, the number of larval instars varies depending on the host plant's identity. Fall webworm (Hyphantria cunea, Drury) is a polyphagous herbivore that feeds on over 450 host plants worldwide. Of the 2 morphotypes (red- and black-head) of fall webworm, the number of instars for the red-head fall webworms has not been characterized. Given its broad diet breadth, fall webworm developmental stages may vary with plant identity. We investigated whether host plant identity affected the number of instars observed during red-head fall webworm development. We measured the head capsules of over 6,000 fall webworm larvae reared on 6 different plants commonly eaten by fall webworms in Colorado. We modeled head capsule widths as Gaussian mixture models, with a Gaussian distribution that corresponded to each instar. We show that our red-head fall webworms varied in number of instars depending on the identity of their host plant upon which they fed. We found that red-head fall webworm exhibited 7 instars on 5 of the host plants and 8 instars on 1 host plant that we studied. Our results for the number of instars for red-head fall webworm are consistent with reports of the number of instars for black-head fall webworm. Our research provides insight into the influence of host plant identity on fall webworm development, which can be used to advance lab and field research of this species.

Potential for biocontrol of Silybum marianum (variegated thistle): 1. Survey of natural enemies in New Zealand

ABSTRACT Silybum marianum (L.) Gaertner (variegated thistle, milk thistle) is one of the worst weeds of hill country pasture in the east coast regions of New Zealand. As part of an initial investigation into the potential for biocontrol, a survey of the phytophagous insects associated with S. marianum in New Zealand was carried out at 16 populations from 29 November to 16 December 2021. Phytophagous associations were determined by collecting insects on the plant and by seedhead dissections. The species collected were grouped into three categories: ‘specialists’, ‘generalists’, and ‘tourists’ that comprised 27.4%, 66.3%, and 6.3% of the total number of specimens collected, respectively. The specialist group was almost entirely comprised of the seedhead weevil, Rhinocyllus conicus (Frölich). While adults of R. conicus were commonly collected on variegated thistle, only 6.5% of seedheads contained larvae of the weevil, indicating that S. marianum is not a common developmental host. Generalist species commonly collected on S. marianum included: Nysius caledoniae Distant, Rhypodes clavicornis (F.), Closterotomus norwegicus (Gmelin) and Nezara virdula (L.). Overall, the field surveys document low levels of attack from generalist herbivores and minimal attack by the specialist weevil, R. conicus. Vacant niches are present on the weed in New Zealand, providing a strong case for introducing specialised biocontrol agents.