Belowground insect herbivory induces systemic volatile emissions that strengthen neighbouring plant resistance aboveground.

Plants transmit ecologically relevant messages to neighbouring plants through chemical cues. For instance, insect herbivory triggers the production of herbivore-induced plant volatiles (HIPVs), which can enhance neighbouring plant defences. HIPVs are emitted from directly damaged plant tissues and from systemic, nondamaged tissues. Although volatile-mediated interplant interactions have been observed both above- and belowground, it remains unknown whether belowground herbivory induces systemic HIPVs aboveground that influence neighbouring plants. To explore how belowground herbivory affects interplant interactions aboveground, we characterised systemic HIPVs from squash induced by belowground striped cucumber beetle (Acalymma vittatum) larval herbivory. We exposed squash 'receiver plants' to systemic HIPVs or volatiles from nondamaged plants. We then measured herbivore resistance by challenging 'receiver plants' with aboveground-feeding herbivores: adult beetles (A. vittatum) or squash bugs (Anasa tristis). We discovered belowground-damaged plants emitted more (E)-ß-ocimene, a key volatile from the systemic HIPV blend, than nondamaged controls, and that exposure to systemic HIPVs enhanced neighbouring plant resistance to aboveground squash bugs, but not adult beetles. Further investigations into the mechanism of interplant interaction revealed ß-ocimene alone can elicit plant resistance against squash bugs. Overall, our findings reveal a novel form of volatile-mediated interactions between plants spanning across aboveground-belowground plant systems.

Aphid-Induced Volatiles and Subsequent Attraction of Natural Enemies Varies among Sorghum Cultivars.

The production of herbivore-induced plant volatiles (HIPVs) is a type of indirect defense used by plants to attract natural enemies and reduce herbivory by insect pests. In many crops little is known about genotypic variation in HIPV production or how this may affect natural enemy attraction. In this study, we identified and quantified HIPVs produced by 10 sorghum (Sorghum bicolor) cultivars infested with a prominent aphid pest, the sorghum aphid (Melanaphis sorghi Theobald). Volatiles were collected using dynamic headspace sampling techniques and identified and quantified using GC-MS. The total amounts of volatiles induced by the aphids did not differ among the 10 cultivars, but overall blends of volatiles differed significantly in composition. Most notably, aphid herbivory induced higher levels of methyl salicylate (MeSA) emission in two cultivars, whereas in four cultivars, the volatile emissions did not change in response to aphid infestation. Dual-choice olfactometer assays were used to determine preference of the aphid parasitoid, Aphelinus nigritus, and predator, Chrysoperla rufilabris, between plants of the same cultivar that were un-infested or infested with aphids. Two aphid-infested cultivars were preferred by natural enemies, while four other cultivars were more attractive to natural enemies when they were free of aphids. The remaining four cultivars elicited no response from parasitoids. Our work suggests that genetic variation in HIPV emissions greatly affects parasitoid and predator attraction to aphid-infested sorghum and that screening crop cultivars for specific predator and parasitoid attractants has the potential to improve the efficacy of biological control.

Plant communication across different environmental contexts suggests a role for stomata in volatile perception.

Plants can detect herbivore-induced plant volatiles (HIPVs) from their damaged neighbours and respond by enhancing or priming their defenses against future herbivore attack. Plant communication and defense priming by volatile cues has been well documented, however, the extent to which plants are able to perceive and respond to these cues across different environmental contexts remains poorly understood. We investigated how abiotic changes that modulate stomatal conductance and/or defense signalling affect the ability of maize plants to perceive HIPVs and respond by priming their defenses. During light exposure, when stomata were open and conditions allowed for defense signal biosynthesis, the individual compounds indole and (Z)-3-hexenyl acetate primed maize defenses. Neither compound primed defenses under environmental conditions that closed stomata and/or altered defense signalling. Moreover, plants were not primed when exposed to indole or (Z)-3-hexenyl acetate in darkness (while stomata were closed) and then subjected to simulated herbivory in the light, to ensure defense induction. The full blend of HIPVs primed maize defenses in light conditions but suppressed defense induction during dark exposure and wounding. These findings indicate that environmental context is important for plant communication and defense priming and suggest that stomata play a role in plant perception of HIPVs.

Root volatile profiles and herbivore preference are mediated by maize domestication, geographic spread, and modern breeding.

MAIN CONCLUSION: Domestication affected the abundances and diversity of maize root volatiles more than northward spread and modern breeding, and herbivore preference for roots was correlated with volatile diversity and herbivore resistance. Studies show that herbivore defenses in crops are mediated by domestication, spread, and breeding, among other human-driven processes. They also show that those processes affected chemical communication between crop plants and herbivores. We hypothesized that (i) preference of the herbivore (Diabrotica virgifera virgifera) larvae for embryonic roots of maize (Zea mays mays) would increase and (ii) root volatile diversity would decrease with the crop's domestication, northward spread to present-day USA, and modern breeding. We used Balsas teosinte (Zea mays parviglumis), Mexican and USA landrace maizes, and US inbred maize lines to test these hypotheses. We found that herbivore preference and volatile diversity increased with maize domestication and northward spread but decreased with modern breeding. Additionally, we found that the abundances of single volatiles did not consistently increase or decrease with maize domestication, spread, and breeding; rather, volatiles grouped per their abundances were differentially affected by those processes, and domestication had the greatest effects. Altogether, our results suggested that: the herbivore's preference for maize roots is correlated with volatile diversity and herbivore resistance; changes in abundances of individual volatiles are evident at the level of volatile groups; and maize domestication, but not spread and breeding, affected the abundances of some green leaf volatiles and sesquiterpenes/sesquiterpenoids. In part, we discussed our results in the context of herbivore defense evolution when resources for plant growth and defense vary across environments. We suggested that variability in relative abundance of volatiles may be associated with their local, functional relevance across wild and agricultural environments.

Consistent signatures of urban adaptation in a native, urban invader ant Tapinoma sessile.

Biological invasions are becoming more prevalent due to the rise of global trade and expansion of urban areas. Ants are among the most prolific invaders with many exhibiting a multiqueen colony structure, dependent colony foundation and reduced internest aggression. Although these characteristics are generally associated with the invasions of exotic ants, they may also facilitate the spread of native ants into novel habitats. Native to diverse habitats across North America, the odorous house ant Tapinoma sessile has become abundant in urban environments throughout the United States. Natural colonies typically have a small workforce, inhabit a single nest, and are headed by a single queen, whereas urban colonies tend to be several orders of magnitude larger, inhabit multiple nests (i.e., polydomy) and are headed by multiple queens (i.e., polygyny). Here, we explore and compare the population genetic and breeding structure of T. sessile within and between urban and natural environments in several localities across its distribution range. We found the social structure of a colony to be a plastic trait in both habitats, although extreme polygyny was confined to urban habitats. Additionally, polydomous colonies were only present in urban habitats, suggesting T. sessile can only achieve supercoloniality within urbanized areas. Finally, we identified strong differentiation between urban and natural populations in each locality and continent-wide, indicating cities may restrict gene flow and exert intense selection pressure. Overall, our study highlights urbanization's influence in charting the evolutionary course for species.

Defense Suppression through Interplant Communication Depends on the Attacking Herbivore Species.

In response to herbivory, plants emit volatile compounds that play important roles in plant defense. Herbivore-induced plant volatiles (HIPVs) can deter herbivores, recruit natural enemies, and warn other plants of possible herbivore attack. Following HIPV detection, neighboring plants often respond by enhancing their anti-herbivore defenses, but a recent study found that herbivores can manipulate HIPV-interplant communication for their own benefit and suppress defenses in neighboring plants. Herbivores induce species-specific blends of HIPVs and how these different blends affect the specificity of plant defense responses remains unclear. Here we assessed how HIPVs from zucchini plants (Cucurbita pepo) challenged with different herbivore species affect resistance in neighboring plants. Volatile "emitter" plants were damaged by one of three herbivore species: saltmarsh caterpillars (Estigmene acrea), squash bugs (Anasa tristis), or striped cucumber beetles (Acalymma vittatum), or were left as undamaged controls. Neighboring "receiver" plants were exposed to HIPVs or control volatiles and then challenged by the associated herbivore species. As measures of plant resistance, we quantified herbivore feeding damage and defense-related phytohormones in receivers. We found that the three herbivore species induced different HIPV blends from squash plants. HIPVs induced by saltmarsh caterpillars suppressed defenses in receivers, leading to greater herbivory and lower defense induction compared to controls. In contrast, HIPVs induced by cucumber beetles and squash bugs did not affect plant resistance to subsequent herbivory in receivers. Our study shows that herbivore species identity affects volatile-mediated interplant communication in zucchini, revealing a new example of herbivore defense suppression through volatile cues.

Chemical Cues from Entomopathogenic Nematodes Vary Across Three Species with Different Foraging Strategies, Triggering Different Behavioral Responses in Prey and Competitors.

Chemical cues play important roles in predator-prey interactions. Semiochemicals can aid predator foraging and alert prey organisms to the presence of predators. Previous work suggests that predator traits differentially influence prey behavior, however, empirical data on how prey organisms respond to chemical cues from predator species with different hunting strategies, and how foraging predators react to cues from potential competitors, is lacking. Furthermore, most research in this area has focused on aquatic and aboveground terrestrial systems, while interactions among belowground, soiling-dwelling organisms have received relatively little attention. Here, we assessed how chemical cues from three species of entomopathogenic nematodes (EPNs), each with a different foraging strategy, influenced herbivore (cucumber beetle) and natural enemy (EPN) foraging behavior. We predicted these cues could serve as chemical indicators of increased predation risk, prey availability, or competition. Our findings revealed that foraging cucumber beetle larvae avoided chemical cues from Heterorhabditis bacteriophora (active-foraging cruiser EPNs), but not Steinernema carpocapsae (ambusher EPNs) or Steinernema riobrave (intermediate-foraging EPNs). In contrast, foraging H. bacteriophora EPNs were attracted to cues produced by the two Steinernema species but not conspecific cues. Notably, the three EPN species produced distinct blends of olfactory cues, with only a few semi-conserved compounds across species. These results indicate that a belowground insect herbivore responds differently to chemical cues from different EPN species, with some EPN species avoiding prey detection. Moreover, the active-hunting EPNs were attracted to heterospecific cues, suggesting these cues indicate a greater probability of available prey, rather than strong interspecific competition.

Asymmetry in Herbivore Effector Responses: Caterpillar Frass Effectors Reduce Performance of a Subsequent Herbivore.

Multiple species of phytophagous insects may co-occur on a plant and while plants can defend themselves from insect herbivory, plant responses to damage by different species and feeding guilds of insects may be asymmetric. Plants can trigger specific responses to elicitors/effectors in insect secretions altering herbivore performance. Recently, maize chitinases present in fall armyworm (FAW, Spodoptera frugiperda) frass were shown to act as effectors suppressing caterpillar-induced defenses in maize while increasing caterpillar performance. We investigated the effect of frass chitinase-mediated suppression of herbivore defenses in maize on the performance and preference of a subsequent insect herbivore from a different feeding guild, corn leaf aphid (Rhopalosiphum maidis). Aphid performance was highest on plants with FAW damage without frass chitinases compared to damaged plants with frass chitinases or undamaged plants. Plant exposure to frass chitinases post FAW damage also altered the production of herbivore-induced volatile compounds compared to damaged, buffer-treated plants. However, aphid preference to damaged, frass chitinase-treated plants was not different from damaged, buffer-treated plants or undamaged plants. This study suggests that frass effector-mediated alteration of plant defenses affects insect herbivores asymmetrically; while it enhances the performance of caterpillars, it suppresses the performance of subsequent herbivores from a different feeding guild.

Exposure of Solidago altissima plants to volatile emissions of an insect antagonist (Eurosta solidaginis) deters subsequent herbivory.

Recent work indicates that plants respond to environmental odors. For example, some parasitic plants grow toward volatile cues from their host plants, and other plants have been shown to exhibit enhanced defense capability after exposure to volatile emissions from herbivore-damaged neighbors. Despite such intriguing discoveries, we currently know relatively little about the occurrence and significance of plant responses to olfactory cues in natural systems. Here we explore the possibility that some plants may respond to the odors of insect antagonists. We report that tall goldenrod (Solidago altissima) plants exposed to the putative sex attractant of a closely associated herbivore, the gall-inducing fly Eurosta solidaginis, exhibit enhanced defense responses and reduced susceptibility to insect feeding damage. In a field study, egg-laying E. solidaginis females discriminated against plants previously exposed to the sex-specific volatile emissions of males; furthermore, overall rates of herbivory were reduced on exposed plants. Consistent with these findings, laboratory assays documented reduced performance of the specialist herbivore Trirhabda virgata on plants exposed to male fly emissions (or crude extracts), as well as enhanced induction of the key defense hormone jasmonic acid in exposed plants after herbivory. These unexpected findings from a classic ecological study system provide evidence for a previously unexplored class of plant-insect interactions involving plant responses to insect-derived olfactory cues.

The volatile emission of Eurosta solidaginis primes herbivore-induced volatile production in Solidago altissima and does not directly deter insect feeding.

BACKGROUND: The induction of plant defenses in response to herbivory is well documented. In addition, many plants prime their anti-herbivore defenses following exposure to environmental cues associated with increased risk of subsequent attack, including induced volatile emissions from herbivore-damaged plant tissues. Recently, we showed in both field and laboratory settings that tall goldenrod plants (Solidago altissima) exposed to the putative sex attractant of a specialist gall-inducing fly (Eurosta solidaginis) experienced less herbivory than unexposed plants. Furthermore, we observed stronger induction of the defense phytohormone jasmonic acid in exposed plants compared to controls. These findings document a novel class of plant-insect interactions mediated by the direct perception, by plants, of insect-derived olfactory cues. However, our previous study did not exclude the possibility that the fly emission (or its residue) might also deter insect feeding via direct effects on the herbivores. RESULTS: Here we show that the E. solidaginis emission does not (directly) deter herbivore feeding on Cucurbita pepo or Symphyotrichum lateriflorum plants--which have no co-evolutionary relationship with E. solidaginis and thus are not expected to exhibit priming responses to the fly emission. We also document stronger induction of herbivore-induced plant volatiles (HIPV) in S. altissima plants given previous exposure to the fly emission relative to unexposed controls. No similar effect was observed in maize plants (Zea mays), which have no co-evolutionary relationship with E. solidaginis. CONCLUSIONS: Together with our previous findings, these results provide compelling evidence that reduced herbivory on S. altissima plants exposed to the emission of male E. solidaginis reflects an evolved plant response to olfactory cues associated with its specialist herbivore and does not involve direct effects of the fly emission on herbivore feeding behavior. We further discuss mechanisms by which the priming of HIPV responses documented here might contribute to enhanced S. altissima defense against galling.

Mortality of resprouting chaparral shrubs after a fire and during a record drought: physiological mechanisms and demographic consequences.

We examined postfire regeneration of chaparral shrubs during an intense drought. This study focused on the demography and physiology of shrub species that resprout from a basal lignotuber following fire. We found significant levels of resprout mortality when intense drought occurred in the year following fire during the period of shrub recovery. Three of the seven sampled resprouting species had the greatest or near greatest levels of mortality ever recorded when compared to previous studies. Most shrub mortality occurred during the drought after individuals had resprouted (i.e. individuals survived fire, resprouted and then subsequently died). Physiological measurements of species with high mortality suggested that resprout stems were highly embolized and xylem hydraulic conductivities were close to zero during the peak of the drought. In addition, lignotubers of two of the three species experiencing high mortality were depleted of starch. Population densities of most shrub species declined after the drought compared with their prefire levels, with the exception of one drought tolerant obligate seeding species. Resprouting shrub species may deplete their carbohydrate reserves during the resprouting process, making them particularly vulnerable to drought because of the need to transpire water to acquire the CO2 that is used to supply energy to a large respiring root system. Drought appears to interact with fire by altering postfire shrub recovery and altering species abundances and composition of chaparral communities.

Phytohormone dynamics associated with gall insects, and their potential role in the evolution of the gall-inducing habit.

While plant galls can be induced by a variety of organisms, insects produce the most diverse and complex galls found in nature; yet, how these galls are formed is unknown. Phytohormones have long been hypothesized to play a key role in gall production, but their exact role, and how they influence galls, has been unclear. Research in the past decade has provided better insight into the role of plant hormones in gall growth and plant defenses. We review and synthesize recent literature on auxin, cytokinins, and abscisic, jasmonic, and salicylic acids to provide a broader understanding of how these phytohormones might effect gall production, help plants defend against galls, and/or allow insects to overcome host-plant defenses. After reviewing these topics, we consider the potential for phytohormones to have facilitated the evolution of insect galls. More specialized research is needed to provide a mechanistic understanding of how phytohormones operate in gall-insect-plant interactions, but current evidence strongly supports phytohormones as key factors determining the success and failure of insect galls.

Identification of a colony- and dose-dependent worker aggregation pheromone in the subterranean termite Reticulitermes virginicus.

Collective behaviors of social insects are often regulated by pheromones. In subterranean termites, some workers forage for and exploit decaying wood for new food resources while forming tunnels from their nest. Colonizing new food resources requires workers to build and disinfect tunnels and chambers inside the nest and ingest decaying wood; therefore subterranean termite colonies should have mechanisms to establish and maintain groups of workers to perform these functions. Recently, an aggregation pheromone was identified in workers of the termite Reticulitermes speratus, which induces quick attraction of nestmate workers and prolonged aggregation to the site of attraction. In this study, we extended this work to another species of Reticulitermes and identified a worker aggregation pheromone in the termite R. virginicus. GC-MS analysis and bioassays demonstrated that this pheromone consists of 3-octanone, 3-octanol, and palmitic acid and shows a colony-specific, dose-dependent attractant response but not an arrestant response. Furthermore, these pheromone components were most likely emitted from the surface of the body. This suggests that aggregation pheromone composition and function differ significantly among termite species, even within the same genus. This study advances our understanding of the regulatory mechanisms of termite aggregation behavior.

Aphelinus nigritus Howard (Hymenoptera: Aphelinidae) Preference for Sorghum Aphid, Melanaphis sorghi (Theobald, 1904) (Hemiptera: Aphididae), Honeydew Is Stronger in Johnson Grass, Sorghum halepense, Than in Grain Sorghum, Sorghum bicolor.

How aphid parasitoids of recent invasive species interact with their hosts can affect the feasibility of biological control. In this study, we focus on a recent invasive pest of US grain sorghum, Sorghum bicolor, the sorghum aphid (SA), Melanaphis sorghi. Understanding this pest's ecology in the grain sorghum agroecosystem is critical to develop effective control strategies. As parasitoids often use aphid honeydew as a sugar resource, and honeydew is known to mediate parasitoid-aphid interactions, we investigated the ability of SA honeydew to retain the parasitoid Aphelinus nigritus. Since SAs in the US have multiple plant hosts, and host-plant diet can modulate parasitoid retention (a major component in host foraging), we measured SA honeydew sugar, organic acid, and amino acid profiles, then assessed via retention time A. nigritus preference for honeydew produced on grain sorghum or Johnson grass, Sorghum halepense. Compared to a water control, A. nigritus spent more time on SA honeydew produced on either host plant. Despite similar honeydew profiles from both plant species, A. nigritus preferred honeydew produced on Johnson grass. Our results suggest the potential for SA honeydew to facilitate augmentation strategies aimed at maintaining A. nigritus on Johnson grass to suppress SAs before grain sorghum is planted.

Improving Natural Enemy Selection in Biological Control through Greater Attention to Chemical Ecology and Host-Associated Differentiation of Target Arthropod Pests.

Host-associated differentiation (HAD) refers to cases in which genetically distinct populations of a species (e.g., herbivores or natural enemies) preferentially reproduce or feed on different host species. In agroecosystems, HAD often results in unique strains or biotypes of pest species, each attacking different species of crops. However, HAD is not restricted to pest populations, and may cascade to the third trophic level, affecting host selection by natural enemies, and ultimately leading to HAD within natural enemy species. Natural enemy HAD may affect the outcomes of biological control efforts, whether classical, conservation, or augmentative. Here, we explore the potential effects of pest and natural enemy HAD on biological control in agroecosystems, with emphases on current knowledge gaps and implications of HAD for selection of biological control agents. Additionally, given the importance of semiochemicals in mediating interactions between trophic levels, we emphasize the role of chemical ecology in interactions between pests and natural enemies, and suggest areas of consideration for biological control. Overall, we aim to jump-start a conversation concerning the relevance of HAD in biological control by reviewing currently available information on natural enemy HAD, identifying challenges to incorporating HAD considerations into biological control efforts, and proposing future research directions on natural enemy selection and HAD.

Adverse effects of inbreeding on the transgenerational expression of herbivore-induced defense traits in Solanum carolinense.

In addition to directly inducing physical and chemical defenses, herbivory experienced by plants in one generation can influence the expression of defensive traits in offspring. Plant defense phenotypes can be compromised by inbreeding, and there is some evidence that such adverse effects can extend to the transgenerational expression of induced resistance. We explored how the inbreeding status of maternal Solanum carolinense plants influenced the transgenerational effects of herbivory on the defensive traits and herbivore resistance of offspring. Manduca sexta caterpillars were used to damage inbred and outbred S. carolinense maternal plants and cross pollinations were performed to produced seeds from herbivore-damaged and undamaged, inbred and outbred maternal plants. Seeds were grown in the greenhouse to assess offspring defense-related traits (i.e., leaf trichomes, internode spines, volatile organic compounds) and resistance to herbivores. We found that feeding by M. sexta caterpillars on maternal plants had a positive influence on trichome and spine production in offspring and that caterpillar development on offspring of herbivore-damaged maternal plants was delayed relative to that on offspring of undamaged plants. Offspring of inbred maternal plants had reduced spine production, compared to those of outbred maternal plants, and caterpillars performed better on the offspring of inbred plants. Both herbivory and inbreeding in the maternal generation altered volatile emissions of offspring. In general, maternal plant inbreeding dampened transgenerational effects of herbivory on offspring defensive traits and herbivore resistance. Taken together, this study demonstrates that inducible defenses in S. carolinense can persist across generations and that inbreeding compromises transgenerational resistance in S. carolinense.

A Beneficial Plant-Associated Fungus Shifts the Balance toward Plant Growth over Resistance, Increasing Cucumber Tolerance to Root Herbivory.

Plants allocate their limited resources toward different physiological processes, dynamically adjusting their resource allocation in response to environmental changes. How beneficial plant-associated microbes influence this allocation is a topic that continues to interest plant biologists. In this study, we examined the effect of a beneficial fungus, Phialemonium inflatum, on investment in growth and anti-herbivore resistance traits in cucumber plants (Cucumis sativus). We inoculated cucumber seeds with P. inflatum spores and measured several growth parameters, including germination rate, above and belowground biomass, and number of flowers. We also examined plant resistance to adult and larval striped cucumber beetles (Acalymma vitattum), and quantified levels of defense hormones in leaves and roots. Our results indicate that P. inflatum strongly enhances cucumber plant growth and reproductive potential. Although fungus treatment did not improve plant resistance to cucumber beetles, inoculated plants were more tolerant to root herbivory, experiencing less biomass reduction. Together, these findings document how a beneficial plant-associated fungus shifts plant investment in growth over herbivore resistance, highlighting the importance of microbes in mediating plant-herbivore interactions. These findings also have important implications for agricultural systems, where beneficial microbes are often introduced or managed to promote plant growth or enhance resistance.

Distinct chemical blends produced by different reproductive castes in the subterranean termite Reticulitermes flavipes.

The production of royal pheromones by reproductives (queens and kings) enables social insect colonies to allocate individuals into reproductive and non-reproductive roles. In many termite species, nestmates can develop into neotenics when the primary king or queen dies, which then inhibit the production of additional reproductives. This suggests that primary reproductives and neotenics produce royal pheromones. The cuticular hydrocarbon heneicosane was identified as a royal pheromone in Reticulitermes flavipes neotenics. Here, we investigated the presence of this and other cuticular hydrocarbons in primary reproductives and neotenics of this species, and the ontogeny of their production in primary reproductives. Our results revealed that heneicosane was produced by most neotenics, raising the question of whether reproductive status may trigger its production. Neotenics produced six additional cuticular hydrocarbons absent from workers and nymphs. Remarkably, heneicosane and four of these compounds were absent in primary reproductives, and the other two compounds were present in lower quantities. Neotenics therefore have a distinct 'royal' blend from primary reproductives, and potentially over-signal their reproductive status. Our results suggest that primary reproductives and neotenics may face different social pressures. Future studies of these pressures should provide a more complete understanding of the mechanisms underlying social regulation in termites.

Foliar herbivory increases sucrose concentration in bracteal extrafloral nectar of cotton.

Cultivated cotton, such as Gossypium hirsutum L., produces extrafloral (EF) nectar on leaves (foliar) and reproductive structures (bracteal) as an indirect anti-herbivore defense. In exchange for this carbohydrate-rich substance, predatory insects such as ants protect the plant against herbivorous insects. Some EF nectar-bearing plants respond to herbivory by increasing EF nectar production. For instance, herbivore-free G. hirsutum produces more bracteal than foliar EF nectar, but increases its foliar EF nectar production in response to herbivory. This study is the first to test for systemically induced changes to the carbohydrate composition of bracteal EF nectar in response to foliar herbivory on G. hirsutum. We found that foliar herbivory significantly increased the sucrose content of bracteal EF nectar while glucose and fructose remained unchanged. Sucrose content is known to influence ant foraging behavior and previous studies of an herbivore-induced increase to EF nectar caloric content found that it led to increased ant activity on the plant. As a follow-up to our finding, ant recruitment to mock EF nectar solutions that varied in sucrose content was tested in the field. The ants did not exhibit any preference for either solution, potentially because sucrose is a minor carbohydrate component in G. hirsutum EF nectar: total sugar content was not significantly affected by the increase in sucrose. Nonetheless, our findings raise new questions about cotton's inducible EF nectar responses to herbivory. Further research is needed to determine whether an herbivore-induced increase in sucrose content is typical of Gossypium spp., and whether it constitutes a corollary of systemic sucrose induction, or a potentially adaptive mechanism which enhances ant attraction to the plant.

Divide and conquer: Multicolonial structure, nestmate recognition, and antagonistic behaviors in dense populations of the invasive ant Brachymyrmex patagonicus.

The ecological success of ants has made them abundant in most environments, yet inter- and intraspecific competition usually limit nest density for a given population. Most invasive ant populations circumvent this limitation through a supercolonial structure, eliminating intraspecific competition through a loss of nestmate recognition and lack of aggression toward non-nestmates. Native to South America, Brachymyrmex patagonicus has recently invaded many locations worldwide, with invasive populations described as extremely large and dense. Yet, in contrast with most invasive ants, this species exhibits a multicolonial structure, whereby each colony occupies a single nest. Here, we investigated the interplay between genetic diversity, chemical recognition, and aggressive behaviors in an invasive population of B. patagonicus. We found that, in its invasive range, this species reaches a high nest density with individual colonies located every 2.5 m and that colony boundaries are maintained through aggression toward non-nestmates. This recognition and antagonism toward non-nestmates is mediated by chemical differentiation between colonies, as different colonies exhibit distinct chemical profiles. We highlighted that the level of aggression between colonies is correlated with their degree of genetic difference, but not their overall chemical differentiation. This may suggest that only a few chemical compounds influence nestmate recognition in this species or that weak chemical differences are sufficient to elicit aggression. Overall, this study demonstrates that invasive ant populations can reach high densities despite a multicolonial structure with strong aggression between colonies, raising questions about the factors underlying their ecological success and mitigating negative consequences of competitive interactions.