Evolutionary Ecology of Multitrophic Interactions between Plants, Insect Herbivores and Entomopathogens

Shikano I., McCarthy E.M., Elderd B.D., Hoover K.

Plant-mediated variations in the outcomes of host-pathogen interactions can strongly affect epizootics and the population dynamics of numerous species, including devastating agricultural pests such as the fall armyworm. Most studies of plant-mediated effects on insect pathogens focus on host mortality, but few have measured pathogen yield, which can affect whether or not an epizootic outbreak occurs. Insects challenged with baculoviruses on different plant species and parts can vary in levels of mortality and yield of infectious stages (occlusion bodies; OBs). We previously demonstrated that soybean genotypes and induced anti-herbivore defenses influence baculovirus infectivity. Here, we used a soybean genotype that strongly reduced baculovirus infectivity when virus was ingested on induced plants (Braxton) and another that did not reduce infectivity (Gasoy), to determine how soybean genotype and induced defenses influence OB yield and speed of kill. These are key fitness measures because baculoviruses are obligate-killing pathogens. We challenged fall armyworm, Spodoptera frugiperda, with the baculovirus S. frugiperda multi-nucleocapsid nucleopolyhedrovirus (SfMNPV) during short or long-term exposure to plant treatments (i.e., induced or non-induced genotypes). Caterpillars were either fed plant treatments only during virus ingestion (short-term exposure to foliage) or from the point of virus ingestion until death (long-term exposure). We found trade-offs of increasing OB yield with slower speed of kill and decreasing virus dose. OB yield increased more with longer time to death and decreased more with increasing virus dose after short-term feeding on Braxton compared with Gasoy. OB yield increased significantly more with time to death in larvae that fed until death on non-induced foliage than induced foliage. Moreover, fewer OBs per unit of host tissue were produced when larvae were fed induced foliage than non-induced foliage. These findings highlight the potential importance of plant effects, even at the individual plant level, on entomopathogen fitness, which may impact epizootic transmission events and host population dynamics.

Shikano I., Rosa C., Tan C., Felton G.W.

It is becoming abundantly clear that the microbes associated with plants and insects can profoundly influence plant-insect interactions. Here, we focus on recent findings and propose directions for future research that involve microbe-induced changes to plant defenses and nutritive quality as well as the consequences of these changes for the behavior and fitness of insect herbivores. Insect (herbivore and parasitoid)-associated microbes can favor or improve insect fitness by suppressing plant defenses and detoxifying defensive phytochemicals. Phytopathogens can influence or manipulate insect behavior and fitness by altering plant quality and defense. Plant-beneficial microbes can promote plant growth and influence plant nutritional and phytochemical composition that can positively or negatively influence insect fitness. Lastly, we suggest that entomopathogens have the potential to influence plant defenses directly as endophytes or indirectly by altering insect physiology.

OLSON G.L., MYERS J.H., HEMERIK L., CORY J.S.

1. When populations of herbivorous insects increase in density, they can alter the quantity or quality of their food. The impacts of diet-related stressors on insect fitness have been investigated singly, but not simultaneously. 2. Foliage quantity and quality of red alder, Alnus rubra, were manipulated together with the presence of non-entomopathogenic phylloplane bacteria to investigate their impacts, singly and in combination, on survival, pupal mass, growth rate, fecundity and egg quality of a cyclic forest insect, the western tent caterpillar, Malacosoma californicum pluviale. 3. Food limitation (half food) had strong negative impacts on all life-history traits. When the larvae were fed continuously, however, neither ingesting phylloplane bacteria nor eating leaves from damaged branches (induced foliage) affected survival. In the half-food treatment, ingesting bacteria further increased mortality, while feeding on induced foliage improved survival. 4. Growth rate and pupal mass of both sexes were reduced for larvae with food limitation compared with continuously fed insects and this was exacerbated when the larvae also ate bacteria-treated leaves. A combination of bacteria and induced foliage also reduced larval growth rate by 5% in the full-food treatment. 5. Fecundity (eggs per egg mass) was 2.7times greater in full-food than in food-limited treatments but neither phylloplane bacteria nor plant induction had an effect. Insects fed induced foliage produced smaller eggs. Overall, there was no evidence of a three-way interaction between the three stressors, although there were negative synergistic effects, primarily between food limitation and the ingestion of phylloplane bacteria.

Brütsch T., Jaffuel G., Vallat A., Turlings T.C., Chapuisat M.

Wood ants fight pathogens by incorporating tree resin with antimicrobial properties into their nests. They also produce large quantities of formic acid in their venom gland, which they readily spray to defend or disinfect their nest. Mixing chemicals to produce powerful antibiotics is common practice in human medicine, yet evidence for the use of such defensive cocktails by animals remains scant. Here, we test the hypothesis that wood ants enhance the antifungal activity of tree resin by treating it with formic acid. In a series of experiments, we document that (i) tree resin had much higher inhibitory activity against the common entomopathogenic fungus Metarhizium brunneum after having been in contact with ants, while no such effect was detected for other nest materials; (ii) wood ants applied significant amounts of endogenous formic and succinic acid on resin and other nest materials; and (iii) the application of synthetic formic acid greatly increased the antifungal activity of resin, but had no such effect when applied to inert glass material. Together, these results demonstrate that wood ants obtain an effective protection against a detrimental microorganism by mixing endogenous and plant-acquired chemical defenses. In conclusion, the ability to synergistically combine antimicrobial substances of diverse origins is not restricted to humans and may play an important role in insect societies.

Palmer‐Young E.C., Sadd B.M., Irwin R.E., Adler L.S.

Floral landscapes comprise diverse phytochemical combinations. Individual phytochemicals in floral nectar and pollen can reduce infection in bees and directly inhibit trypanosome parasites. However, gut parasites of generalist pollinators, which consume nectar and pollen from many plant species, are exposed to phytochemical combinations. Interactions between phytochemicals could augment or decrease effects of single compounds on parasites. Using a matrix of 36 phytochemical treatment combinations, we assessed the combined effects of two floral phytochemicals, eugenol and thymol, against four strains of the bumblebee gut trypanosome Crithidia bombi. Eugenol and thymol had synergistic effects against C. bombi growth across seven independent experiments, showing that the phytochemical combination can disproportionately inhibit parasites. The strength of synergistic effects varied across strains and experiments. Thus, the antiparasitic effects of individual compounds will depend on both the presence of other phytochemicals and parasite strain identity. The presence of synergistic phytochemical combinations could augment the antiparasitic activity of individual compounds for pollinators in diverse floral landscapes.

Acevedo F.E., Peiffer M., Tan C., Stanley B.A., Stanley A., Wang J., Jones A.G., Hoover K., Rosa C., Luthe D., Felton G.

Mechanical damage caused by insect feeding along with components present in insect saliva and oral secretions are known to induce jasmonic acid–mediated defense responses in plants. This study investigated the effects of bacteria from oral secretions of the fall armyworm Spodoptera frugiperda on herbivore-induced defenses in tomato and maize plants. Using culture-dependent methods, we identified seven different bacterial isolates belonging to the family Enterobacteriacea from the oral secretions of field-collected caterpillars. Two isolates, Pantoea ananatis and Enterobacteriaceae-1, downregulated the activity of the plant defensive proteins polyphenol oxidase and trypsin proteinase inhibitors (trypsin PI) but upregulated peroxidase (POX) activity in tomato. A Raoultella sp. and a Klebsiella sp. downregulated POX but upregulated trypsin PI in this plant species. Conversely, all of these bacterial isolates upregulated the expression of the herbivore-induced maize proteinase inhibitor (mpi) gene in maize. Plant treatment with P. ananatis and Enterobacteriaceae-1 enhanced caterpillar growth on tomato but diminished their growth on maize plants. Our results highlight the importance of herbivore-associated microbes and their ability to mediate insect plant interactions differently in host plants fed on by the same herbivore.

Shikano I., Shumaker K.L., Peiffer M., Felton G.W., Hoover K.

Baculoviruses are food-borne microbial pathogens that are ingested by insects on contaminated foliage. Oxidation of plant-derived phenolics, activated by insect feeding, can directly interfere with infections in the gut. Since phenolic oxidation is an important component of plant resistance against insects, baculoviruses are suggested to be incompatible with plant defences. However, plants among and within species invest differently in a myriad of chemical and physical defences. Therefore, we hypothesized that among eight soybean genotypes, some genotypes would be able to maintain both high resistance against an insect pest and high efficacy of a baculovirus. Soybean constitutive (non-induced) and jasmonic acid (JA)-induced (anti-herbivore response) resistance was measured against the fall armyworm Spodoptera frugiperda (weight gain, leaf consumption and utilization). Indicators of phenolic oxidation were measured as foliar phenolic content and peroxidase activity. Levels of armyworm mortality inflicted by baculovirus (SfMNPV) did not vary among soybean genotypes when the virus was ingested with non-induced foliage. Ingestion of the virus on JA-induced foliage reduced armyworm mortality, relative to non-induced foliage, on some soybean genotypes. Baculovirus efficacy was lower when ingested with foliage that contained higher phenolic content and defensive properties that reduced armyworm weight gain and leaf utilization. However, soybean genotypes that defended the plant by reducing consumption rate and strongly deterred feeding upon JA-induction did not reduce baculovirus efficacy, indicating that these defences may be more compatible with baculoviruses to maximize plant protection. Differential compatibility of defence traits with the third trophic level highlights an important cost/trade-off associated with plant defence strategies.

Chung S.H., Scully E.D., Peiffer M., Geib S.M., Rosa C., Hoover K., Felton G.W.

Herbivore associated bacteria are vital mediators of plant and insect interactions. Host plants play an important role in shaping the gut bacterial community of insects. Colorado potato beetles (CPB; Leptinotarsa decemlineata) use several Solanum plants as hosts in their natural environment. We previously showed that symbiotic gut bacteria from CPB larvae suppressed jasmonate (JA)-induced defenses in tomato. However, little is known about how changes in the bacterial community may be involved in the manipulation of induced defenses in wild and cultivated Solanum plants of CPB. Here, we examined suppression of JA-mediated defense in wild and cultivated hosts of CPB by chemical elicitors and their symbiotic bacteria. Furthermore, we investigated associations between the gut bacterial community and suppression of plant defenses using 16 S rRNA amplicon sequencing. Symbiotic bacteria decreased plant defenses in all Solanum hosts and there were different gut bacterial communities in CPB fed on different host plants. When larvae were reared on different hosts, defense suppression differed among host plants. These results demonstrate that host plants influence herbivore gut bacterial communities and consequently affect the herbivore’s ability to manipulate JA-mediated plant defenses. Thus, the presence of symbiotic bacteria that suppress plant defenses might help CPB adapt to host plants.

Shikano I., Cory J.S.

Infection by parasites can alter the feeding behaviour of hosts. Some animals seek out substances that can therapeutically clear infections (self-medication), some may seek out resources to recoup resources lost while fighting off infection (compensatory feeding) and others may be manipulated to ingest substances that benefit parasite fitness (parasite manipulation of host). Recent studies have indicated that pathogen-challenged insects can self-medicate by increasing their protein intake relative to carbohydrate, which is thought to act by boosting the insect's immune response. However, increased protein intake could also be due to compensatory feeding or pathogen manipulation of the host, and a rigorous examination of all four of the testable predictions, which is necessary for verifying self-medication behaviour, has not been conducted. The therapeutic behaviour must (1) only be employed by infected individuals and (2) alleviate the potential fitness loss of the infected individual. (3) If an uninfected individual engages in the behaviour, they suffer a decrease in fitness, and lastly, (4) the parasite cannot benefit from the behaviour. In response to baculovirus-challenge (AcMNPV) at 24°C, the cabbage looper, Trichoplusia ni, increased proportional protein intake, by increasing protein intake rather than decreasing carbohydrate intake. Increased protein intake did not benefit virus fitness, but it also did not increase the probability of host survival. Increased proportional protein intake did not occur in response to TnSNPV-challenge at 24°C or in response to AcMNPV-challenge at a higher temperature (32°C), indicating that the virus-induced change in nutrient intake depends on virus identity and temperature. Since virus-challenged T. ni did not show the typical costs associated with infection, the altered nutrient intake is likely to be a compensatory response. Understanding the motivation behind pathogen-induced changes in feeding behaviour could have significant implications for determining its importance for species interactions at multiple trophic levels.

Couture J.J., Mason C.J., Habeck C.W., Lindroth R.L.

Animals have several strategies to contend with nutritionally poor diets, including compensatory consumption and enhanced food utilization efficiencies. Plants produce a diversity of defense compounds that affect the ability of herbivores to utilize these strategies in response to variation in food nutritional quality. Little is known, however, about effects of allelochemicals on herbivores utilizing integrated behavioral and morphological responses to reduced food quality. Our objectives were to (1) examine how variation in diet nutritional quality influences compensatory responses of a generalist insect herbivore, and (2) determine how plant defenses affect these processes. Gypsy moth (Lymantria dispar) larvae were administered one of nine combinations of diet having low, moderate, or high nutritional quality and 0, 2, or 4 % purified aspen (Populus tremuloides) salicinoids. We quantified larval growth, consumption, frass production, and biomass allocation to midgut tissue over a 4-day bioassay. In the absence of salicinoids, larvae compensated for reduced nutritional quality and maintained similar growth across all diets through increased consumption, altered midgut biomass allocation, and improved processing efficiencies. Dietary salicinoids reduced larval consumption, midgut biomass allocation, digestive efficiencies, and growth at all nutritional levels, but the effect size was more pronounced when larvae were fed nutritionally suboptimal diets. Our findings demonstrate that integrated behavioral and morphological compensatory responses to reduced food quality are affected by plant defenses, ultimately limiting compensatory responses and reducing larval performance.

Shikano I., Hua K.N., Cory J.S.

Invertebrate hosts that survive pathogen challenge can produce offspring that are more resistant to the same pathogen via immune priming, thereby improving the fitness of their offspring in the same pathogen environment. Most evidence for immune priming comes from exposure to bacteria and there are limited data on other groups of pathogens. Poor parental nutrition has also been shown to result in the transgenerational transfer of pathogen resistance and increased immunocompetence. Here, we combine exposure to an insect DNA virus with a change in the parental diet to examine both parental costs and transgenerational immune priming. We challenged the cabbage looper, Trichoplusia ni, with a low dose of the baculovirus, Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and altered dietary protein to carbohydrate ratio (p:c ratio) after virus exposure. Insects fed a low protein diet had lower haemolymph protein concentrations, and exhibited costs of smaller pupae and slower development, while survivors of virus challenge developed more slowly, irrespective of p:c ratio, and those that were virus-challenged and fed on a low protein diet showed a reduction in haemocyte density. In addition, AcMNPV-challenged parents laid fewer eggs earlier in egg laying although egg size was the same as for unchallenged parents. There was no evidence for increased resistance to AcMNPV (immune priming) or changes in haemocyte number (as proxy for constitutive cellular immunity) in the offspring either as a result of parental AcMNPV-challenge or low dietary p:c ratio. Therefore, although pathogen-challenge and nutritional changes can affect host development and reproduction, this does not necessarily translate into transgenerational immune priming. Our findings contrast with an earlier study on another type of baculovirus, a granulovirus, where immune priming was suggested. This indicates that transgenerational immune priming is not universal in invertebrates and is likely to depend on the host-pathogen system, or the level of pathogen exposure and the type of dietary manipulation. Identifying whether immune priming or transgenerational effects are relevant in field populations, remains a challenge.

Wan N.-., Jiang J.-., Li B.

Previous studies have shown that the infectivity of baculovirus to herbivores is affected by phytochemicals ingested during the acquisition of viral inoculum on the foliage of host plants. Here, we measured the effects of 14 host plant species on the infectivity of Spodoptera exigua nucleopolyhedrovirus (SeNPV) to its larvae. The order of the LD50 values of SeNPV among the host plants was Ipomoea aquatica > Brassica oleracea > Raphanus sativus > Amaranthus tricolor > Spinacia oleracea > Vigna unguiculata > Solanum melongena > Capsicum annuum > Apium graveolens > Allium fistulosum > Lactuca sativa > Brassica chinensis > Zea mays > Glycine max, with 940.1 ± 2.26, 424.0 ± 0.60, 295.2 ± 1.13, 147.3 ± 0.63, 138.6 ± 0.22, 119.9 ± 0.07, 119.8 ± 0.02, 109.2 ± 0.18, 104.8 ± 0.62, 102.1 ± 0.66, 97.9 ± 0.22, 89.9 ± 0.32, 79.0 ± 0.13 and 64.0 ± 0.38 OBs per larva, respectively, and the values of mean time to death of virus-infected larvae were 6.21 ± 0.11, 7.12 ± 0.10, 7.33 ± 0.21, 6.97 ± 0.02, 7.06 ± 0.01, 7.29 ± 0.03, 7.32 ± 0.05, 7.07 ± 0.08, 7.24 ± 0.11, 7.09 ± 0.13, 7.50 ± 0.06, 7.23 ± 0.01, 7.30 ± 0.02 and 7.19 ± 0.07 days, respectively. The mean time to death of larvae decreased with increasing viral dose, and corrected mortality decreased as the larval mean time to death increased. These findings have significance for understanding the effects of host plants on the infectivity of baculovirus to noctuids.

Lacey L.A., Grzywacz D., Shapiro-Ilan D.I., Frutos R., Brownbridge M., Goettel M.S.

The development and use of entomopathogens as classical, conservation and augmentative biological control agents have included a number of successes and some setbacks in the past 1years. In this forum paper we present current information on development, use and future directions of insect-specific viruses, bacteria, fungi and nematodes as components of integrated pest management strategies for control of arthropod pests of crops, forests, urban habitats, and insects of medical and veterinary importance. Insect pathogenic viruses are a fruitful source of microbial control agents (MCAs), particularly for the control of lepidopteran pests. Most research is focused on the baculoviruses, important pathogens of some globally important pests for which control has become difficult due to either pesticide resistance or pressure to reduce pesticide residues. Baculoviruses are accepted as safe, readily mass produced, highly pathogenic and easily formulated and applied control agents. New baculovirus products are appearing in many countries and gaining an increased market share. However, the absence of a practical in vitro mass production system, generally higher production costs, limited post application persistence, slow rate of kill and high host specificity currently contribute to restricted use in pest control. Overcoming these limitations are key research areas for which progress could open up use of insect viruses to much larger markets. A small number of entomopathogenic bacteria have been commercially developed for control of insect pests. These include several Bacillus thuringiensis sub-species, Lysinibacillus (Bacillus) sphaericus, Paenibacillus spp. and Serratia entomophila. B. thuringiensis sub-species kurstaki is the most widely used for control of pest insects of crops and forests, and B. thuringiensis sub-species israelensis and L. sphaericus are the primary pathogens used for control of medically important pests including dipteran vectors. These pathogens combine the advantages of chemical pesticides and MCAs: they are fast acting, easy to produce at a relatively low cost, easy to formulate, have a long shelf life and allow delivery using conventional application equipment and systemics (i.e. in transgenic plants). Unlike broad spectrum chemical pesticides, B. thuringiensis toxins are selective and negative environmental impact is very limited. Of the several commercially produced MCAs, B. thuringiensis (Bt) has more than 50% of market share. Extensive research, particularly on the molecular mode of action of Bt toxins, has been conducted over the past two decades. The Bt genes used in insect-resistant transgenic crops belong to the Cry and vegetative insecticidal protein families of toxins. Bt has been highly efficacious in pest management of corn and cotton, drastically reducing the amount of broad spectrum chemical insecticides used while being safe for consumers and non-target organisms. Despite successes, the adoption of Bt crops has not been without controversy. Although there is a lack of scientific evidence regarding their detrimental effects, this controversy has created the widespread perception in some quarters that Bt crops are dangerous for the environment. In addition to discovery of more efficacious isolates and toxins, an increase in the use of Bt products and transgenes will rely on innovations in formulation, better delivery systems and ultimately, wider public acceptance of transgenic plants expressing insect-specific Bt toxins. Fungi are ubiquitous natural entomopathogens that often cause epizootics in host insects and possess many desirable traits that favor their development as MCAs. Presently, commercialized microbial pesticides based on entomopathogenic fungi largely occupy niche markets. A variety of molecular tools and technologies have recently allowed reclassification of numerous species based on phylogeny, as well as matching anamorphs (asexual forms) and teleomorphs (sexual forms) of several entomopathogenic taxa in the Phylum Ascomycota. Although these fungi have been traditionally regarded exclusively as pathogens of arthropods, recent studies have demonstrated that they occupy a great diversity of ecological niches. Entomopathogenic fungi are now known to be plant endophytes, plant disease antagonists, rhizosphere colonizers, and plant growth promoters. These newly understood attributes provide possibilities to use fungi in multiple roles. In addition to arthropod pest control, some fungal species could simultaneously suppress plant pathogens and plant parasitic nematodes as well as promote plant growth. A greater understanding of fungal ecology is needed to define their roles in nature and evaluate their limitations in biological control. More efficient mass production, formulation and delivery systems must be devised to supply an ever increasing market. More testing under field conditions is required to identify effects of biotic and abiotic factors on efficacy and persistence. Lastly, greater attention must be paid to their use within integrated pest management programs; in particular, strategies that incorporate fungi in combination with arthropod predators and parasitoids need to be defined to ensure compatibility and maximize efficacy. Entomopathogenic nematodes (EPNs) in the genera Steinernema and Heterorhabditis are potent MCAs. Substantial progress in research and application of EPNs has been made in the past decade. The number of target pests shown to be susceptible to EPNs has continued to increase. Advancements in this regard primarily have been made in soil habitats where EPNs are shielded from environmental extremes, but progress has also been made in use of nematodes in above-ground habitats owing to the development of improved protective formulations. Progress has also resulted from advancements in nematode production technology using both in vivo and in vitro systems; novel application methods such as distribution of infected host cadavers; and nematode strain improvement via enhancement and stabilization of beneficial traits. Innovative research has also yielded insights into the fundamentals of EPN biology including major advances in genomics, nematode-bacterial symbiont interactions, ecological relationships, and foraging behavior. Additional research is needed to leverage these basic findings toward direct improvements in microbial control.

Malik D., Kumar S., Sindhu S.S.

Christensen T., Smilanich A.M., Carper A., Peechatt V., Bowers M.D., Forister M.L., Teglas M.B., Hurtado P., Dyer L.A.

Although infectious diseases play a critical role in population regulation, our knowledge of complex drivers of disease for insects is limited. We conducted a field study on Baltimore checkerspot caterpillars ( Euphydryas phaeton ), chemical specialists on plants containing iridoid glycosides (IGs), to investigate the roles of host plant, phytochemistry, ontogeny and spatial associations in determining viral prevalence. We analysed individuals for viral presence and loads, quantified leaf IG concentrations from their native and novel host plants, and sequestered IGs in caterpillars. We found proximate caterpillar groups had greater similarity in infection prevalence, with areas of high prevalence indicating viral hotspots. Underlying variation in host plant chemistry corresponded to differences in viral prevalence. Furthermore, we used structural equation modeling to examine causal drivers of infection prevalence and loads. Advanced ontogeny was associated with increased viral prevalence and loads, as well as decreased sequestration of IGs. Infection loads were lower on the novel host plant, but prevalence was slightly higher, partially explained by decreased sequestration of IGs. Altogether, our findings reveal that spatial proximity, ontogeny, host plant species and secondary phytochemistry can all contribute to structuring infection risk, and thus offer insight into causal drivers of disease prevalence in complex plant–insect systems.

Sanaei E., de Roode J.C.

ABSTRACTIn this comprehensive exploration, we delve into the pivotal role of host plants in shaping the intricate interactions between herbivorous insects and their pathogens. Recent decades have seen a surge in studies that demonstrate that host plants are crucial drivers of the interactions between insects and pathogens, providing novel insights into the direct and indirect interactions that shape tri‐trophic interactions. These studies have built on a wide range of pathogens, from viruses to bacteria, and from protozoans to fungi. We summarise these studies, and discuss the mechanisms of plant‐mediated insect resistance to infection, ranging from the toxicity of plant chemicals to pathogens to enhancement of anti‐pathogen immune responses, and modulation of the insect's microbiome. Although we provide evidence for the roles of all these mechanisms, we also point out that the majority of existing studies are phenomenological, describing patterns without addressing the underlying mechanisms. To further our understanding of these tri‐trophic interactions, we therefore urge researchers to design their studies to enable them specifically to distinguish the mechanisms by which plants affect insect susceptibility to pathogens.

Jones A.G., Shikano I., Mason C.J., Peiffer M., Felton G.W., Hoover K.

Baculoviruses are a group of entomopathogenic viruses that are important natural enemies of insects, particularly lepidopteran larvae. An important component of baculovirus transmission efficiency is the frequency with which hosts encounter patchily distributed virions on plants. Little is known about the ecology and bacterial composition of virus-killed cadavers. We used a baculovirus and host Trichoplusia ni caterpillars to study the effects of virus-killed cadavers on tomato plant defenses and T. ni behavior. We also compared bacterial communities associated with virus-killed and uninfected (freeze-killed) cadavers and found that there was no significant difference in community composition and membership between tomato-fed virus-killed or freeze-killed cadavers. Comparison of virus-killed cadavers from two separate experiments revealed significant differences in bacterial community composition, suggesting that host plant could play a more important role in shaping bacterial communities than virus infection status. Culture-dependent plating indicated that virus-killed cadavers had significantly higher bacterial titers compared with uninfected cadavers. We found that virus-killed cadavers suppressed polyphenol oxidase activity, an important plant defense protein, in mechanically damaged plants, but not in plants damaged by herbivory. Although cadavers did not influence plant defenses induced by feeding damage inflicted by healthy or infected T. ni, this study provides the first evidence that baculoviruses could influence plant defenses through host cadavers. When applied to intact plants, neither virus-killed or freeze-killed cadavers influenced T. ni oviposition, larval choice, or larval consumption, indicating these insects did not discriminate cadaver cues. Virus-killed cadavers could play important roles in mediating interactions between plants, herbivores, and other trophic levels, with potential implications for viral transmission dynamics.

López-Ferber M., Caballero P., Williams T.

Baculoviruses can naturally regulate lepidopteran populations and are used as biological insecticides. The genetic diversity of these viruses affects their survival and efficacy in pest control. For nucleopolyhedroviruses, occlusion-derived virions and the occlusion body facilitate the transmission of groups of genomes, whereas this is not the case for granuloviruses. We review the evidence for baculovirus genetic diversity in the environment, in the host insect, and in occlusion bodies and virions. Coinfection allows defective genotypes to persist through complementation and results in the pseudotyping of virus progeny that can influence their transmissibility and insecticidal properties. Genetic diversity has marked implications for the development of pest resistance to virus insecticides. We conclude that future research is warranted on the physical segregation of genomes during virus replication and on the independent action of virions during infection. We also identify opportunities for studies on the transmission of genetic diversity and host resistance to viruses.

Monica S.S., Rajanikanth P., Duraimurugan P., Ameer Basha S., Srinivasa Chary D.

Frattini A., González-Martínez R.M., García J.M., Minchev Z., Pozo M.J., Flors V., Crava C.M., Herrero S.

Thirumeni M.A., Yashkamal K., Mohamed R.R., Shivakumar M.S.

Agriculture productivity today faces many challenges in meeting the requirements of a growing population. Climate change and pest damage are two of the primary constraints for an increase in agricultural productivity. Damage to agricultural crops primarily occurs due to insect pests. Chemical insecticides are used for the control of insect pests on crops and for stored grain insect pests. These chemical insecticides are harmful for humans and other nontarget organisms and also result in ecosystem damage. After a few exposures, the target insect pests evolve to overcome the toxic effects of these insecticides, making them redundant. In order to overcome these drawbacks, a few alternate and environmentally friendly methods have been tested. These include the sterile insect male technique, the use of microbes and their metabolites, the bioprospecting of secondary metabolites from plants, and gene editing technology. Although each method has its pros and cons, the use of microorganisms and their metabolites has emerged as a low-cost, environmentally sustainable method for the control of insect pests. Among the microbes used in insect control, insect pathogenic fungi hold potential to be used as an inexpensive alternative to chemical insecticides. Only a few of these have been commercialized. The popularity of these entomopathogenic fungi is low, as fungal pesticides have a relatively short shelf life; their virulence gets low after long storage, and spore germination is affected by the high temperature and low humidity. Recent studies have shown that modifying the media composition can increase the virulence and shelf life of fungi. Given the diversity of entomopathogenic fungi and their secondary metabolites, there is a huge potential for developing highly effective biopesticides for insect control.

Falcon-Brindis A., Villanueva R.T.

Pest management on hemp is still in its infancy, and biological control options are limited. Helicoverpa zea (corn earworm) is one of the key pests of hemp cultivated outdoors, especially on cultivars grown for cannabinoids and grain. In a three-year study, we assessed the effect of diet on the performance of H. zea and its tachinid parasitoids. Parasitized (bearing fly eggs) and unparasitized (without eggs) H. zea larvae were fed on hemp flowers or an artificial diet. Five tachinid species parasitized H. zea larvae, but the most abundant species were Winthemia rufopicta (68.8%) and Lespesia aletiae (28.3%). Overall, 55.2% of H. zea larvae bearing tachinid eggs died, while the mortality of unparasitized larvae reached 24.7%. The success of tachinids increased by 2-fold when the host larvae were fed on an artificial diet. Our results demonstrated that high protein food (artificial diet), intensity of parasitism, and caterpillar size play a role in the fitness of both the herbivores (H. zea) and its tachinid parasitoids. These findings have important implications for understanding biological control mechanisms and open new insights into the impact of landscape variation on plant-herbivore-parasitoid interactions. This study contains supporting evidence that makes both Winthemia rufopicta and Lespesia aletiae excellent candidates for biological control programs against H. zea, a key pest of hemp in the United States.

Wang J., Jiang J., Fan N., Wu H., Ji X., Wan N., Hou Y.

Plant secondary metabolites are crucial in affecting the interactions between insect herbivores and entomoviruses. However, there is limited knowledge regarding the impact of such metabolites on the susceptibility of insect herbivores to entomoviruses. In this study, we adopted the allicin, caterpillars (Spodoptera exigua) and nucleopolyhedrovirus (SeMNPV) as a system, and found that allicin significantly increased the mortality of S. exigua larvae infected with SeMNPV by 36.03–59.45% when infected with the virus at a concentration of 2.12 × 103 OB·mL−1. Furthermore, NPV-infected larvae together treated with allicin inhibited the growth and development of larvae, comparing to individual NPV-infected larvae. Notably, we observed a significant enrichment of differentially expressed genes involved in the cytochrome P450-mediated metabolism pathway between the NPV-infected and allicin combined with NPV-treated groups. The silencing of CYP340AA1 through RNA interference significantly increased the mortality of larvae infected with SeMNPV. This investigation indicates that allicin might be a potential candidate for improving the performance of the NPV against insect herbivores and identifies that CYP340AA1 gene is important in this process.

Dhiman V.K., Singh D., Dhiman V.K., Pandey M., Sharma A., Pandey H., Verma S.K.

Plants are surrounded by the biotic factors around their ecosystem. Their interaction with insects is one such important factor that shapes the habitat around them. Insects may be beneficial or pests depending on their activity in the microclimate they live with the flora around them. The plants are provided with physical strength, chemical responses, and physiological changes with time to adapt to the herbivory nature of insects. Some insects live in mutual and take food and shelter for survival, providing plants with defense and predatory response to pest populations. Others cause tissue damage, triggering responses like chemical secretion that either attract pests for toxic feeders or repel them. The physical architecture of tissue structures also provides the advantage to plants to cope with damage and regeneration. Some defense mechanism varies with species of plants and are specific based on their interaction with insects.

Armstrong K.M., Uyeda J., Shikano I.

Fear associated with perceived predation risk can elicit behavioral and physiological changes in animals, including insects. Diamondback moth (DBM), Plutella xylostella, larvae are known to wriggle violently backwards and even fall off plants when attacked by predators and parasitoids. In the laboratory, the DBM larvae exposed to the specialist parasitoid Diadegma insulare were found to move to lower quality, older foliage, which negatively affected their development. In Hawaii, by far the dominant parasitoid species of DBM is the generalist Cotesia vestalis, with many sampling efforts often only yielding this species. Thus, in this study, we investigated the influence of C. vestalis on the behavior of DBM larvae. We focused on the distribution of DBM on cabbage plants, from the youngest to oldest leaves (leaf position 1–12, respectively), in the presence of C. vestalis in the lab and field. We found that in the lab, DBM larvae showed a clear preference for younger leaves (leaf positions 3–5). However, when C. vestalis was present, regardless of whether the parasitoids could touch the DBM, larvae were less likely to be on those preferred leaf positions. This change in larval distribution was also replicated when potted cabbage plants containing DBM larvae were placed in cabbage fields. Changes in herbivore feeding sites, whether mediated by natural enemies or other environmental factors, could have significant implications, not only for herbivore fitness, but also for economic damage to crops based on the value of plant parts affected.

Ramírez-Ordorica A., Adame-Garnica S.G., Ramos-Aboites H.E., Winkler R., Macías-Rodríguez L.

Beauveria bassiana is an entomopathogenic fungus that parasitizes and kills insects. The role of volatile organic compounds (VOCs) emitted by B. bassiana acting as semiochemicals during its interaction with lepidopterans is poorly explored. Here, we studied the effect of VOCs from B. bassiana and 3-methylbutanol (as a single compound) on the feeding behavior of L2 larvae of Spodoptera frugiperda in sorghum plants. Additionally, we assessed whether fungal VOCs induce chemical modifications in the plants that affect larval food preferences. Metabolomic profiling of plant tissues was performed by mass spectrometry and bioassays in a dual-choice olfactometer. The results showed that the larval feeding behavior was affected by the B. bassiana strain AI2, showing that the insect response is strain-specific. Furthermore, 80 µg of 3-methylbutanol affected the number of bites. The larval feeding choice was dependent on the background context. Fragment spectra and a matching precursor ion mass of 165.882 m/z enabled the putative identification of 4-coumaric acid in sorghum leaves exposed to fungal VOCs, which may be associated with larval deterrent responses. These results provide valuable insights into the bipartite interaction of B. bassiana with lepidopterans through VOC emission, with the plant as a mediator of the interaction.

Zhang B., Yang W., He Q., Chen H., Che B., Bai X.

As an indispensable part of insects, intestinal symbiotic bacteria play a vital role in the growth and development of insects and their adaptability. Rhoptroceros cyatheae, the main pest of the relict plant Alsophila spinulosa, poses a serious threat to the development of the A. spinulosa population. In the present study, 16S rDNA and internal transcribed spacer high-throughput sequencing techniques were used to analyze the structure of intestinal microbes and the diversity of the insect feeding on two different plants, as well as the similarities between the intestinal microorganisms of R. cyatheae. The dominant bacteria of leaf endophytes were also compared based on the sequencing data. The results showed that Proteobacteria, Firmicutes, and Actinobacteria were the dominant phyla of intestinal bacteria, and Ascomycota was the dominant phylum of intestinal fungi. Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, Methylobacterium-Methylorubrum, and Enterococcus were the dominant genera in the intestine of R. cyatheae feeding on two plants, and the relative abundance was significantly different between the two groups. Candida was the common dominant genus of intestinal fungi in the two groups, and no significant difference was observed in its abundance between the two groups. This showed that compared with the intestinal fungi of R. cyatheae, the abundance of the intestinal bacteria was greatly affected by food. The common core microbiota between the microorganisms in A. spinulosa leaves and the insect gut indicated the presence of a microbial exchange between the two. The network correlation diagram showed that the gut microbes of R. cyatheae feeding on Gymnosphaera metteniana were more closely related to each other, which could help the host to better cope with the adverse external environment. This study provides a theoretical basis for the adaptation mechanism of R. cyatheae and a new direction for the effective prevention and control of R. cyatheae.

Wang J., Zhang S., Kong J., Chang J.

BackgroundThe plant secondary metabolites (PSMs), as important plant resistance indicators, are important targets for screening plant insect resistance breeding. In this study, we aimed to investigate whether the population of Zeuzera coffeae (ZC) is affected by different varieties of Carya illinoinensis PSMs content. At the same time, the structure and function of the gut microbiome of ZC were also analyzed in relation to different pecan varieties.MethodsWe counted the populations of ZC larvae in four pecan varieties and determined the content of four types of PSMs. The structure and function of the larval gut microbiota were studied in connection to the number of larvae and the content of PSMs. The relationships were investigated between larval number, larval gut microbiota, and PSM content.ResultsWe found that the tannins, total phenolics, and total saponins of 4 various pecans PSMs stifled the development of the ZC larval population. The PSMs can significantly affect the diversity and abundance of the larval gut microbiota. Enrichment of ASV46 (Pararhizobium sp.), ASV994 (Olivibacter sp.), ASV743 (Rhizobium sp.), ASV709 (Rhizobium sp.), ASV671 (Luteolibacter sp.), ASV599 (Agrobacterium sp.), ASV575 (Microbacterium sp.), and ASV27 (Rhizobium sp.) in the gut of larvae fed on high-resistance cultivars was positively associated with their tannin, total saponin, and total phenolic content. The results of the gut microbiome functional prediction for larvae fed highly resistant pecan varieties showed that the enriched pathways in the gut were related to the breakdown of hazardous chemicals.ConclusionOur findings provide further evidence that pecan PSMs influence the structure and function of the gut microbiota, which in turn affects the population stability of ZC. The study’s findings can serve as a theoretical foundation for further work on selecting ZC-resistant cultivars and developing green management technology for ZC.