The plant toxin 4-methylsulfinylbutyl isothiocyanate decreases herbivore performance and modulates cellular and humoral immunity.

Insect herbivores frequently encounter plant defense molecules, but the physiological and ecological consequences for their immune systems are not fully understood. The majority of studies attempting to relate levels of plant defensive chemistry to herbivore immune responses have used natural population or species-level variation in plant defensive chemistry. Yet, this potentially confounds the effects of plant defense chemistry with other potential plant trait differences that may affect the expression of herbivore immunity. We used an artificial diet containing known quantities of a plant toxin (4-methylsulfinylbutyl isothiocyanate; 4MSOB-ITC or ITC, a breakdown product of the glucosinolate glucoraphanin upon herbivory) to explicitly explore the effects of a plant toxin on the cellular and humoral immune responses of the generalist herbivore Trichoplusia ni (Lepidoptera: Noctuidae) that frequently feeds on glucosinolate-containing plants. Caterpillars feeding on diets with high concentrations of ITC experienced reduced survivorship and growth rates. High concentrations of ITC suppressed the appearance of several types of hemocytes and melanization activity, which are critical defenses against parasitic Hymenoptera and microbial pathogens. In terms of T. ni humoral immunity, only the antimicrobial peptide (AMP) genes lebocin and gallerimycin were significantly upregulated in caterpillars fed on diets containing high levels of ITC relative to caterpillars that were provided with ITC-free diet. Surprisingly, challenging caterpillars with a non-pathogenic strain of Escherichia coli resulted in the upregulation of the AMP gene cecropin. Feeding on high concentrations of plant toxins hindered caterpillar development, decreased cellular immunity, but conferred mixed effects on humoral immunity. Our findings provide novel insights into the effects of herbivore diet composition on insect performance demonstrating the role of specific plant defense toxins that shape herbivore immunity and trophic interactions.

Trait-based approaches to predicting biological control success: challenges and prospects.

Identifying traits that are associated with success of introduced natural enemies in establishing and controlling pest insects has occupied researchers and biological control practitioners for decades. Unfortunately, consistent general relationships have been difficult to detect, preventing a priori ranking of candidate biological control agents based on their traits. We summarise previous efforts and propose a series of potential explanations for the lack of clear patterns. We argue that the quality of current datasets is insufficient to detect complex trait-efficacy relationships and suggest several measures by which current limitations may be overcome. We conclude that efforts to address this elusive issue have not yet been exhausted and that further explorations are likely to be worthwhile.

Oviposition Preference and Performance of a Specialist Herbivore Is Modulated by Natural Enemies, Larval Odors, and Immune Status.

Insect herbivores frequently must balance host plant quality and the risk of attack by their natural enemies when making oviposition decisions. Yet, which factor is more important remains unresolved in plant-insect ecology. Here, we report the oviposition preference and larval performance of the brassicaceous specialist Plutella xylostella, in the context of plant quality (cabbage Brassica oleracea vs. mustard B. juncea) and associated natural enemies. Despite the greater larval weight and adult lifespan on cabbage, ovipositing females strongly preferred mustard. Both the egg parasitoid Trichogrammatoidea bactrae and the larval ectoparasitoid Bracon brevicornis are more likely to attack P. xylostella that feed on cabbage; thus, mustard represents enemy-reduced space from these two parasitoids. However, larval diet had no impact on the parasitism rate of specialist Cotesia vestalis. Feeding on mustard improved larval immune responses. The total hemocyte number, diversity, and phenoloxidase activity were higher in mustard-fed larvae which increased their survival against the entomopathogen, Bacillus thuringiensis. Interestingly, host plants altered the larval body odor profile. Mustard-fed larvae emitted allyl isothiocyanate (AITC) and butyl isothiocyanate (BITC) while cabbage-fed larvae emitted dimethyl disulphide (DMDS) and dimethyl trisulphide (DMTS) that served as short-range cues for larval parasitoids. For B. brevicornis, host body odor guided oviposition choice was crucial as their fitness was affected by the host larval diet. Although C. vestalis showed a clear preference towards volatiles emitted by mustard fed larvae, their fitness was unaltered. Taken together, our results illustrate that P. xylostella prefers to lay eggs on mustard plants providing enemy-reduced space from some, but not all, natural enemies.

Extrinsic Inter- and Intraspecific Competition in Parasitoid Wasps.

The diverse ecology of parasitoids is shaped by extrinsic competition, i.e., exploitative or interference competition among adult females and males for hosts and mates. Adult females use an array of morphological, chemical, and behavioral mechanisms to engage in competition that may be either intra- or interspecific. Weaker competitors are often excluded or, if they persist, use alternate host habitats, host developmental stages, or host species. Competition among adult males for mates is almost exclusively intraspecific and involves visual displays, chemical signals, and even physical combat. Extrinsic competition influences community structure through its role in competitive displacement and apparent competition. Finally, anthropogenic changes such as habitat loss and fragmentation, invasive species, pollutants, and climate change result in phenological mismatches and range expansions within host-parasitoid communities with consequent changes to the strength of competitive interactions. Such changes have important ramifications not only for the success of managed agroecosystems, but also for natural ecosystem functioning.

A Chromosome-Level Genome Assembly of the Parasitoid Wasp, Cotesia glomerata (Hymenoptera: Braconidae).

Hymenopterans make up about 20% of all animal species, but most are poorly known and lack high-quality genomic resources. One group of important, yet understudied hymenopterans are parasitoid wasps in the family Braconidae. Among this understudied group is the genus Cotesia, a clade of ~1,000 species routinely used in studies of physiology, ecology, biological control, and genetics. However, our ability to understand these organisms has been hindered by a lack of genomic resources. We helped bridge this gap by generating a high-quality genome assembly for the parasitoid wasp, Cotesia glomerata (Braconidae; Microgastrinae). We generated this assembly using multiple sequencing technologies, including Oxford Nanopore, whole-genome shotgun sequencing, and 3D chromatin contact information (HiC). Our assembly is one of the most contiguous, complete, and publicly available hymenopteran genomes, represented by 3,355 scaffolds with a scaffold N50 of ~28 Mb and a BUSCO score of ~99%. Given the genome sizes found in closely related species, our genome assembly was ~50% larger than expected, which was apparently induced by runaway amplification of 3 types of repetitive elements: simple repeats, long terminal repeats, and long interspersed nuclear elements. This assembly is another step forward for genomics across this hyperdiverse, yet understudied order of insects. The assembled genomic data and metadata files are publicly available via Figshare (https://doi.org/10.6084/m9.figshare.13010549).

Effects of Landscape Composition on Wheat Stem Sawfly (Hymenoptera: Cephidae) and Its Associated Braconid Parasitoids.

Several agroecological and integrated pest management strategies focus on landscape management to increase complexity and foster biodiversity. However, landscape complexity does not always enhance biological control and in some cases may lead to increased pest populations. We examined the prevalence of two Bracon parasitoids, Bracon cephi Gahan and Bracon lissogaster Muesebeck (Hymenoptera: Braconidae), and their host the wheat stem sawfly Cephus cinctus Norton, a major pest of wheat. We assessed the degree of noncrop and crop host plant use and responses to landscape composition. We found no instances of parasitism by either Bracon species in our three-year, statewide winter wheat survey but found small populations of Bracon in noncrop landscapes throughout eastern and western Colorado. We used model selection to examine how local (500 m scale) and landscape (5 km scale) cover of suitable noncrop and crop habitats potentially affects abundances of Bracon and wheat stem sawfly. Our best fit model for wheat stem sawfly suggests that a decrease in noncrop cover at the landscape scale leads to an increase in wheat stem sawfly infestation. Our best fit model for Bracon parasitism suggests an increase in wheat cover at the local level results in the greatest increase in the odds of parasitism by either species of Bracon. Herbaceous cover at local and landscape scales were also significant predictors of Bracon parasitism. The results of this study suggest that pest and natural enemies respond differently to landscape composition and these responses should be evaluated before management decisions are made.

Associational effects of plant ontogeny on damage by a specialist insect herbivore.

Intraspecific variation in plant traits is a major cause of variation in herbivore feeding and performance. Plant defensive traits change as a plant grows, such that ontogeny may account for a substantial portion of intraspecific trait variation. We tested how the ontogenic stage of an individual plant, of an individual in the context of its neighboring plants, and of a patch of plants with mixed or uniform stages affect plant-herbivore interactions. To do this, we conducted an experimental study of the interactions between Lepidium draba, a perennial brassicaceous weed, and Plutella xylostella, a common herbivore of L. draba. We found that L. draba foliar glucosinolates, secondary metabolites often implicated in defense, decreased in concentration with plant age. In single-stage patches, herbivores performed similarly on L. draba plants of different ages. Furthermore, we found no difference in the cumulative performance of herbivores reared on mixed- or even-staged patches of L. draba. However, in mixed-stage patches, the damage experienced by a focal plant depended on the stage of neighboring plants, suggesting a preference hierarchy of the herbivore among plant stages. In our study, the amount of herbivory depended on the ontogenic neighborhood in which the plant grew. However, from the herbivore's perspective, variation in plant ontogenic stage was unimportant to its success in terms of feeding rate and final weight.

Generalising indirect defence and resistance of plants.

Indirect defence, the adaptive top-down control of herbivores by plant traits that enhance predation, is a central component of plant-herbivore interactions. However, the scope of interactions that comprise indirect defence and associated ecological and evolutionary processes has not been clearly defined. We argue that the range of plant traits that mediate indirect defence is much greater than previously thought, and we further organise major concepts surrounding their ecological functioning. Despite the wide range of plant traits and interacting organisms involved, indirect defences show commonalities when grouped. These categories are based on whether indirect defences boost natural enemy abundance via food or shelter resources, or, alternatively, increase natural enemy foraging efficiency via information or alteration of habitat complexity. The benefits of indirect defences to natural enemies should be further explored to establish the conditions in which indirect defence generates a plant-natural enemy mutualism. By considering the broader scope of plant-herbivore-natural enemy interactions that comprise indirect defence, we can better understand plant-based food webs, as well as the evolutionary processes that have shaped them.

Population- and Species-Based Variation of Webworm-Parasitoid Interactions in Hogweeds (Heracelum spp.) in the Netherlands.

In three Dutch populations of the native small hogweed (Heracleum sphondylium L. [Apiales: Apiaceae]), and one of the invasive giant hogweed (H. mantegazzianum Sommeier & Levier [Apiales: Apiaceae]), interactions between a specialist herbivore, the parsnip webworm (Depressaria radiella), and its associated parasitoids were compared during a single growing season. We found host plant species-related differences in the abundance of moth pupae, the specialist polyembryonic endoparasitoid, Copidosoma sosares, the specialist pupal parasitoid, Barichneumon heracliana, and a potential hyperparasitoid of C. sosares, Tyndaricus scaurus Walker (Hymenoptera: Encyrtidae). Adult D. radiella body mass was similar across the three small hogweed populations, but moths and their pupal parasitoid B. heracliana were smaller when developing on giant than on small hogweeds where the two plants grew in the same locality (Heteren). Mixed-sex and all-male broods of C. sosares were generally bigger than all-female broods. Furthermore, adult female C. sosares were larger than males and adult female mass differed among the three small hogweed populations. The frequency of pupal parasitism and hyperparasitism also varied in the different H. sphondylium populations. These results show that short-term (intra-seasonal) effects of plant population on multitrophic insects are variable among different species in a tightly linked food chain.

Ecological dissociation and re-association with a superior competitor alters host selection behavior in a parasitoid wasp.

Interspecific competition for limited resources can drive ecological specialization and trait expression. Organisms released from intense competition may exploit a broader range of resources, but if reunited with stronger competitors, survivorship may depend on foraging behaviors that reduce competition. We compared the host selection behavior of the parasitoid Cotesia glomerata from two North American populations that differ in their association with Cotesia rubecula, a superior competitor. Both parasitoids originate from Europe and attack the imported cabbageworm (a.k.a. small cabbage white) Pieris rapae, but C. glomerata was introduced into North America almost a century before C. rubecula. After re-association in North America, C. rubecula has displaced C. glomerata in several regions, but not in other regions. Host selection was measured in female C. glomerata from Maryland (MD) where it coexists with C. rubecula, and in conspecifics from Colorado (CO) where C. rubecula is absent. Unparasitized and C. rubecula-parasitized P. rapae hosts were used in choice tests to examine whether C. glomerata host selection behavior differed based on the population's association history with C. rubecula. We found that C. glomerata from MD had a higher likelihood of avoiding hosts parasitized by C. rubecula (and thus avoiding competition) than did wasps from CO. The ability of C. glomerata to avoid hosts parasitized by C. rubecula may facilitate coexistence in MD; whereas, the lack of discrimination in CO populations of C. glomerata naïve to C. rubecula could contribute to the displacement of C. glomerata were C. rubecula to enter the same habitat.

Plant toxins and parasitoid trophic ecology.

Parasitoids (parasitic wasps) are ubiquitous components of nearly all communities containing plant-insect herbivore associations. Plant toxin defenses against herbivores may also affect higher trophic levels by directly (e.g., plant toxins encountered in host hemolymph) or indirectly (e.g., plant toxins reduce host size/quality or alter the host's immunity against parasitoids). Yet, whether parasitoids structure plant-herbivore interactions remains relatively understudied. Nevertheless, recent meta-analyses and empirical work emphasize the importance of parasitoids in structuring interactions among lower trophic levels. Two promising areas of research are particularly ripe for future exploration: a) the potential for microbes to alter the interactions among plants, insect herbivores, and parasitoids, and b) the effects of climate change on phenological (mis)matches among trophic levels.

Ant-like Traits in Wingless Parasitoids Repel Attack from Wolf Spiders.

A recent study showed that a wingless parasitoid, Gelis agilis, exhibits a suite of ant-like traits that repels attack from wolf spiders. When agitated, G. agilis secreted 6-methyl-5-hepten-2-one (sulcatone), which a small number of ant species produce as an alarm/panic pheromone. Here, we tested four Gelis parasitoid species, occurring in the same food chain and microhabitats, for the presence of sulcatone and conducted two-species choice bioassays with wolf spiders to determine their degree of susceptibility to attack. All four Gelis species, including both winged and wingless species, produced sulcatone, whereas a closely related species, Acrolyta nens, and the more distantly related Cotesia glomerata, did not. In two-choice bioassays, spiders overwhelmingly rejected the wingless Gelis species, preferring A. nens and C. glomerata. However, spiders exhibited no preference for either A. nens or G. areator, both of which are winged. Wingless gelines exhibited several ant-like traits, perhaps accounting for the reluctance of spiders to attack them. On the other hand, despite producing sulcatone, the winged G. areator more closely resembles other winged cryptines like A. nens, making it harder for spiders to distinguish between these two species. C. glomerata was also preferred by spiders over A. nens, suggesting that other non-sulcatone producing cryptines nevertheless possess traits that make them less attractive as prey. Phylogenetic reconstruction of the Cryptinae reveals that G. hortensis and G. proximus are 'sister'species, with G. agilis, and G.areator in particular evolving along more distant trajectories. We discuss the possibility that wingless Gelis species have evolved a suite of ant-like traits as a form, of mimicry to repel predators on the ground.

Lessons from the multitudes: insights from polyembryonic wasps for behavioral ecology.

Even for parasitic Hymenoptera, polyembryonic wasps are unusual creatures. Two features in particular, allow for novel exploration of major questions in behavioral ecology: the production of multiple offspring per egg and, in some species, the production of a soldier caste. Because final brood sizes of polyembryonic species are not constrained by trade-offs between current and future parental reproductive effort, we can clearly examine the selective forces at play that drive the balance between the number of offspring and their body size. Polyembryony also provides excellent opportunities to compare the performance of identical genotypes under different environmental conditions. Finally, polyembryonic species can provide unique tests of how genetic conflicts at multiple levels are resolved.

Variation in Plant Defense Suppresses Herbivore Performance.

Defensive variability of crops and natural systems can alter herbivore communities and reduce herbivory [1, 2]. However, it is still unknown how defense variability translates into herbivore suppression. Nonlinear averaging and constraints in physiological tracking (also more generally called time-dependent effects) are the two mechanisms by which defense variability might impact herbivores [3, 4]. We conducted a set of experiments manipulating the mean and variability of a plant defense, showing that defense variability does suppress herbivore performance and that it does so through physiological tracking effects that cannot be explained by nonlinear averaging. While nonlinear averaging predicted higher or the same herbivore performance on a variable defense than on an invariable defense, we show that variability actually decreased herbivore performance and population growth rate. Defense variability reduces herbivore performance in a way that is more than the average of its parts. This is consistent with constraints in physiological matching of detoxification systems for herbivores experiencing variable toxin levels in their diet and represents a more generalizable way of understanding the impacts of variability on herbivory [5]. Increasing defense variability in croplands at a scale encountered by individual herbivores can suppress herbivory, even if that is not anticipated by nonlinear averaging.

Host Plants of the Wheat Stem Sawfly (Hymenoptera: Cephidae).

Wheat stem sawfly (Cephus cinctus Norton) is a pest of economic importance across much of the wheat (Triticum aestivum L.)-growing areas of the western Great Plains of North America as well as an ecologically important insect owing to its wide range of grass hosts. Little research has been published regarding the noncultivated native and invasive grasses attacked by this insect. Knowledge of the complete host range of C. cinctus can inform future research about potential new sources of genetic resistance, improve understanding of the biology and spread of natural enemies, and better define this insect's role in grassland and agricultural systems. The aim of this review is to compile a checklist of reported host plants of C. cinctus and present data from an extensive survey of noncultivated hosts used by C. cinctus.

Balancing selection maintains sex determining alleles in multiple-locus complementary sex determination.

Hymenopteran species in which sex is determined through a haplo-diploid mechanism known as complementary sex determination (CSD) are vulnerable to a unique form of inbreeding depression. Diploids heterozygous at one or more CSD loci develop into females but diploids homozygous at all loci develop into diploid males, which are generally sterile or inviable. Species with multiple polymorphic CSD loci (ml-CSD) may have lower rates of diploid male production than species with a single CSD locus (sl-CSD), but it is not clear if polymorphism is consistently maintained at all loci. Here, we assess the rate of diploid male production in a population of Cotesia rubecula, a two-locus CSD parasitoid wasp species, approximately 20 years after the population was introduced for biological control. We show that diploid male production dropped from 8-13% in 2005 and 2006 to 3-4% by 2015. We also show from experimental crosses that the population maintained polymorphism at both CSD loci in 2015. We use theory and simulations to show that balancing selection on all CSD alleles promotes polymorphism at several loci in ml-CSD populations. Our study supports the hypothesis that ml-CSD populations have lower diploid male production and are more likely to persist than comparable sl-CSD populations.

MODIS Imagery Improves Pest Risk Assessment: A Case Study of Wheat Stem Sawfly (Cephus cinctus, Hymenoptera: Cephidae) in Colorado, USA.

Wheat stem sawfly (Cephus cinctus Norton, Hymenoptera: Cephidae) has long been a significant insect pest of spring, and more recently, winter wheat in the northern Great Plains. Wheat stem sawfly was first observed infesting winter wheat in Colorado in 2010 and, subsequently, has spread rapidly throughout wheat production regions of the state. Here, we used maximum entropy modeling (MaxEnt) to generate habitat suitability maps in order to predict the risk of crop damage as this species spreads throughout the winter wheat-growing regions of Colorado. We identified environmental variables that influence the current distribution of wheat stem sawfly in the state and evaluated whether remotely sensed variables improved model performance. We used presence localities of C. cinctus and climatic, topographic, soils, and normalized difference vegetation index and enhanced vegetation index data derived from Moderate Resolution Imaging Spectroradiometer (MODIS) imagery as environmental variables. All models had high performance in that they were successful in predicting suitable habitat for C. cinctus in its current distribution in eastern Colorado. The enhanced vegetation index for the month of April improved model performance and was identified as a top contributor to MaxEnt model. Soil clay percent at 0-5 cm, temperature seasonality, and precipitation seasonality were also associated with C. cinctus distribution in Colorado. The improved model performance resulting from integrating vegetation indices in our study demonstrates the ability of remote sensing technologies to enhance species distribution modeling. These risk maps generated can assist managers in planning control measures for current infestations and assess the future risk of C. cinctus establishment in currently uninfested regions.

Integration of Plant Defense Traits with Biological Control of Arthropod Pests: Challenges and Opportunities.

Crop plants exhibit a wide diversity of defensive traits and strategies to protect themselves from damage by herbivorous pests and disease. These defensive traits may be naturally occurring or artificially selected through crop breeding, including introduction via genetic engineering. While these traits can have obvious and direct impacts on herbivorous pests, many have profound effects on higher trophic levels, including the natural enemies of herbivores. Multi-trophic effects of host plant resistance have the potential to influence, both positively and negatively, biological control. Plant defense traits can influence both the numerical and functional responses of natural enemies; these interactions can be semiochemically, plant toxin-, plant nutrient-, and/or physically mediated. Case studies involving predators, parasitoids, and pathogens of crop pests will be presented and discussed. These diverse groups of natural enemies may respond differently to crop plant traits based on their own unique biology and the ecological niches they fill. Genetically modified crop plants that have been engineered to express transgenic products affecting herbivorous pests are an additional consideration. For the most part, transgenic plant incorporated protectant (PIP) traits are compatible with biological control due to their selective toxicity to targeted pests and relatively low non-target impacts, although transgenic crops may have indirect effects on higher trophic levels and arthropod communities mediated by lower host or prey number and/or quality. Host plant resistance and biological control are two of the key pillars of integrated pest management; their potential interactions, whether they are synergistic, complementary, or disruptive, are key in understanding and achieving sustainable and effective pest management.

Hidden risks and benefits of natural enemy-mediated indirect effects.

Polyphagous natural enemies can mediate a variety of indirect interactions between resource populations. Such indirect interactions are often reciprocally negative (i.e. apparent competition), but the sign of effects between resource populations can be any combination of positive (+), negative (-), or neutral (0). In this article we focus on parasitoids to illustrate the importance of natural enemy-mediated indirect interactions in predicting risk and efficacy in biological control. We review recent findings to illustrate how an improved understanding of parasitoid behavioral ecology may increase model accuracy.

Indirect plant-parasitoid interactions mediated by changes in herbivore physiology.

In occupying an intermediate trophic position, herbivorous insects serve a vital link between plants at the base of the food chain and parasitoids at the top. Although these herbivore-mediated indirect plant-parasitoid interactions are well-documented, new studies have uncovered previously undescribed mechanisms that are fundamentally changing how we view tri-trophic relationships. In this review we highlight recent advances in this field focusing on both plant-driven and parasitoid-driven outcomes that flow up and down the trophic web, respectively. From the bottom-up, plant metabolites can impact parasitoid success by altering host immune function; however, few have considered the potential effects of other plant defense strategies such as tolerance on parasitoid ecology and behavior. From the top-down, parasitoids have long been considered plant bodyguards, but in reality the consequences of parasitism for herbivory rates and induction of plant defensive chemistry are far more complicated with cascading effects on community-level interactions.