Aboveground Herbivores Research Articles

Characterizing rhizosphere microbial communities associated with tolerance to aboveground herbivory in wild and domesticated tomatoes.

Root-associated microbial communities are well known for their ability to prime and augment plant defenses that reduce herbivore survival or alter behavior (i.e., resistance). In contrast, the role root microbes play in plant tolerance to herbivory, an evolutionarily sustainable alternative to resistance, is overlooked. In this study, we aimed to expand our limited understanding of what role rhizosphere microbial communities play in supporting tolerance to insect damage. Using domesticated tomatoes and their wild ancestors (Solanum spp.), we first documented how tobacco hornworm (Manduca sexta) herbivory impacted tomato fruit production in order to quantify plant tolerance. We then characterized the bacterial and fungal rhizosphere communities harbored by high and low tolerance plants. Wild tomatoes excelled at tolerating hornworm herbivory, experiencing no significant yield loss despite 50% leaf area removal. Their domesticated counterparts, on the other hand, suffered 26% yield losses under hornworm herbivory, indicating low tolerance. Ontogeny (i.e., mid- vs. late-season sampling) explained the most variation in rhizosphere community structure, with tomato line, tolerance, and domestication status also shaping rhizosphere communities. Fungal and bacterial community traits that associated with the high tolerance line include (1) high species richness, (2) relatively stable community composition under herbivory, and (3) the relative abundance of taxa belonging to Stenotrophomonas, Sphingobacterium, and Sphingomonas. Characterizing tolerance-associating microbiomes may open new avenues through which plant defenses are amended in pest management, such as plant breeding efforts that enhance crop recruitment of beneficial microbiomes.

Legume plant defenses and nutrients mediate indirect interactions between soil rhizobia and chewing herbivores

Soil bacteria that associate with plant roots promote host vigor. Legume plants form mutualisms with rhizobial bacteria, and legumes grown with rhizobia have more nutrients and defenses than those grown without rhizobia. However, few studies have tested how stressors such as herbivores affect soil rhizobia, and the mechanisms mediating these interactions. Here we tested reciprocal interactions between a chewing herbivore, Sitona lineatus (pea leaf weevil), and Pisum sativum (pea) plants grown with or without rhizobia (Rhizobium leguminosarum biovar. viciae), and the plant-defense and nutritional mechanisms mediating these interactions. We hypothesized that plants grown with rhizobia would have less feeding from S. lineatus due to greater expression of phytohormones or physical defenses. We also predicted that herbivory might impede the mutualism between P. sativum and rhizobia. Our experiments showed that leaf defoliation by S. lineatus was indeed lowest on plants with rhizobia. Plants grown with rhizobia had increased gene transcript expression associated with hormone-related defense (jasmonic acid, ethylene, abscisic acid) as well as physical and antioxidant-related defense, which may explain reduced feeding by S. lineatus. Conversely, S. lineatus feeding reduced the number of root nodules and nodule fresh weight, suggesting a disruption of the symbiosis between plants and rhizobia. Our study shows that aboveground herbivores can engage in mutually antagonistic interactions with soil microbes that are mediated through multiple plant-mediated pathways.

Glyphosate residues in soil can modify plant resistance to herbivores through changes in leaf quality.

Glyphosate is the most widely used non-selective herbicide in the world. Glyphosate residues in soil can affect plant quality by modifying plant physiology, hormonal pathways and traits, with potential consequences for plants' interactions with herbivores. We explored these indirect effects in the context of plant-herbivore interactions in a perennial, nitrogen-fixing herb. We quantified leaf herbivory for glyphosate-exposed and control plants grown in phosphorus-fertilized and non-fertilized soils, and assessed the impacts of glyphosate treatment on traits related to plant resistance against herbivores (leaf trichome density, leaf mass per area) and performance (aboveground biomass, root:shoot ratio, nodule number, nodule activity). Moreover, we conducted a laboratory feeding experiment to compare the palatability of leaves from glyphosate-exposed and control plants to a generalist mollusc herbivore. Herbivore damage and intensity in situ increased during the growing season regardless of glyphosate or phosphorus treatment. Glyphosate treatment reduced leaf trichome density but had no effect on the other plant traits considered. Herbivore damage was negatively associated with leaf trichome density. The feeding experiment revealed no difference in the feeding probability of mollusc herbivores between glyphosate-exposed and control plants. However, there was an interaction between glyphosate treatment and initial leaf area for leaf consumption by herbivores: leaf consumption increased with increasing leaf area in both groups, but at a lower rate for glyphosate-exposed plants than for control plants. Our results show that glyphosate residues in soil have the potential to indirectly affect aboveground herbivores through changes in leaf quality, which may have mixed consequences for folivore damage.

Specialist root herbivore modulates plant transcriptome and downregulates defensive secondary metabolites in a brassicaceous plant.

Summary Plants face attackers aboveground and belowground. Insect root herbivores can lead to severe crop losses, yet the underlying transcriptomic responses have rarely been studied.We studied the dynamics of the transcriptomic response of Brussels sprouts (Brassica oleracea var. gemmifera) primary roots to feeding damage by cabbage root fly larvae (Delia radicum), alone or in combination with aboveground herbivory by cabbage aphids (Brevicoryne brassicae) or diamondback moth caterpillars (Plutella xylostella). This was supplemented with analyses of phytohormones and the main classes of secondary metabolites; aromatic, indole and aliphatic glucosinolates.Root herbivory leads to major transcriptomic rearrangement that is modulated by aboveground feeding caterpillars, but not aphids, through priming soon after root feeding starts. The root herbivore downregulates aliphatic glucosinolates. Knocking out aliphatic glucosinolate biosynthesis with CRISPR‐Cas9 results in enhanced performance of the specialist root herbivore, indicating that the herbivore downregulates an effective defence.This study advances our understanding of how plants cope with root herbivory and highlights several novel aspects of insect–plant interactions for future research. Further, our findings may help breeders develop a sustainable solution to a devastating root pest.

Belowground and aboveground herbivory differentially affect the transcriptome in roots and shoots of maize.

Plants recognize and respond to feeding by herbivorous insects by upregulating their local and systemic defenses. While defense induction by aboveground herbivores has been well studied, far less is known about local and systemic defense responses against attacks by belowground herbivores. Here, we investigated and compared the responses of the maize transcriptome to belowground and aboveground mechanical damage and infestation by two well‐adapted herbivores: the soil‐dwelling western corn rootworm Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) and the leaf‐chewing fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae). In responses to both herbivores, maize plants were found to alter local transcription of genes involved in phytohormone signaling, primary and secondary metabolism. Induction by real herbivore damage was considerably stronger and modified the expression of more genes than mechanical damage. Feeding by the corn rootworm had a strong impact on the shoot transcriptome, including the activation of genes involved in defense and development. By contrast, feeding by the fall armyworm induced only few transcriptional changes in the roots. In conclusion, feeding by a leaf chewer and a root feeder differentially affects the local and systemic defense of maize plants. Besides revealing clear differences in how maize plants respond to feeding by these specialized herbivores, this study reveals several novel genes that may play key roles in plant–insect interactions and thus sets the stage for in depth research into the mechanism that can be exploited for improved crop protection.Significance statementExtensive transcriptomic analyses revealed a clear distinction between the gene expression profiles in maize plants upon shoot and root attack, locally as well as distantly from the attacked tissue. This provides detailed insights into the specificity of orchestrated plant defense responses, and the dataset offers a molecular resource for further genetic studies on maize resistance to herbivores and paves the way for novel strategies to enhance maize resistance to pests.

Arbuscular mycorrhizal fungi mediate how plant herbivory history influences herbivore performance

Abstract Herbivore performance is influenced by plant host quality and chemical defences, both of which are influenced by belowground symbionts of plants and herbivory history. The previous herbivory may suppress or facilitate subsequent herbivore performance through changes in plant chemistry and tissue quality. Likewise, arbuscular mycorrhizal fungi (AM fungi) can alter both plant quality and plant responses to aboveground herbivory, as mycorrhizal‐mediated alterations of the nutritional budget may alter leaf nutritional quality, chemical profiles, and available leaf surface area. Despite evidence that both herbivory history and AM fungi can independently affect herbivore performance, few studies directly test both at the same time. Here, the role of AM fungi and herbivory history on the performance of a pest insect (Spodoptera frugiperda) on a common forb (Solidago altissima) grown with different AM fungal additions (no AM fungi, single species, multispecies) was directly tested. Evidence that the effect of herbivory history on herbivore performance depended on the AM fungal environment of the plant was found. Specifically, the composition of the AM fungal additions altered the directionality of herbivore performance on plants with and without prior herbivory. Without added AM fungi, herbivores demonstrated a classically suppressed growth rate on grazed plants. With a single species AM fungi added, herbivores performed the same on host plants, regardless of herbivory history. With diverse AM fungi added, herbivore growth increased on grazed plants. Mycorrhizal‐mediated changes in plant chemical profiles did not explain herbivore performance. These results emphasise the need for direct tests of mycorrhizal diversity on plant‐insect interactions.

Mechanisms of Isothiocyanate Detoxification in Larvae of Two Belowground Herbivores, Delia radicum and D. floralis (Diptera: Anthomyiidae).

Like aboveground herbivores, belowground herbivores are confronted with multiple plant defense mechanisms including complex chemical cocktails in plant tissue. Roots and shoots of Brassicaceae plants contain the two-component glucosinolate (GSL)-myrosinase defense system. Upon cell damage, for example by herbivore feeding, toxic and pungent isothiocyanates (ITCs) can be formed. Several aboveground-feeding herbivores have developed biochemical adaptation strategies to overcome the GSL-ITC defenses of their host plant. Whether belowground herbivores feeding on Brassica roots possess similar mechanisms has received little attention. Here, we analyze how two related belowground specialist herbivores detoxify the GSL-ITC defenses of their host plants. The larvae of the fly species Delia radicum and D. floralis are common pests and specialized herbivores on the roots of Brassicaceae. We used chemical analyses (HPLC-MS/MS and HPLC-UV) to examine how the GSL-ITC defense system is metabolized by these congeneric larvae. In addition, we screened for candidate genes involved in the detoxification process using RNAseq and qPCR. The chemical analyses yielded glutathione conjugates and amines. This indicates that both species detoxify ITCs using potentially the general mercapturic acid pathway, which is also found in aboveground herbivores, and an ITC-specific hydrolytic pathway previously characterized in microbes. Performance assays confirmed that ITCs negatively affect the survival of both species, in spite of their known specialization to ITC-producing plants and tissues, whereas ITC breakdown products are less toxic. Interestingly, the RNAseq analyses showed that the two congeneric species activate different sets of genes upon ITC exposure, which was supported by qPCR data. Based on our findings, we conclude that these specialist larvae use combinations of general and compound-specific detoxification mechanisms with differing efficacies and substrate preferences. This indicates that combining detoxification mechanisms can be an evolutionarily successful strategy to handle plant defenses in herbivores.

Foliar Herbivory Reduces Rhizosphere Fungal Diversity and Destabilizes the Co-occurrence Network.

Insect herbivores can adversely impact terrestrial plants throughout ontogeny and across various ecosystems. Simultaneously, the effects of foliar herbivory may extend belowground, to the soil microbial community. However, the responses in terms of the diversity, assembly, and stability of rhizosphere fungi to aboveground herbivory remain understudied. Here, using high-throughput sequencing, the effects of foliar insect herbivory on rhizosphere fungal microbes were investigated in a common garden experiment that manipulated herbivory intensity and time from herbivore removal. The number of observed fungal species was reduced by a greater herbivory intensity, with some species evidently sensitive to herbivory intensity and time since herbivore removal. Rhizofungal assembly processes were altered by both herbivory intensity and time since herbivore removal. Further, we found evidence that both factors strongly influenced fungal community stability: a high intensity of herbivory coupled with a shorter time since herbivore removal resulted in low stability. These results suggest that foliar herbivory can adversely alter fungal diversity and stability, which would in turn be harmful for plant health. Fortunately, the effect seems to gradually diminish with time elapsed after herbivore removal. Our findings provide a fresh, in-depth view into the roles of rhizofungi in enhancing the adaption ability of plants under environmental stress.

Rapid transfer of C and N excreted by decomposer soil animals to plants and above-ground herbivores

The role of soil decomposer animals in nutrient cycling is commonly seen as indirect, slow and cumulative, through grazing and engineering effects on soil microbes and their mineralization activities. Here, we show that nitrogen (N) and carbon (C) movement from living soil animals to plants and herbivores can be extremely rapid. In two complementary experiments under laboratory and field conditions, we used stable isotope tracers to follow nutrient transfer from earthworms to soil, wheat seedlings, and aphids. Earthworm-derived 15N and 13C was acquired by aphids after 2 h under laboratory conditions, and 15N after 24 h in the field. Since cleansed earthworms with voided guts were introduced, the main source of this 15N and 13C likely was their highly labelled cutaneous mucus. Based on the rapid tracer appearance in a phloem-feeding herbivore (whose principal N source is free amino acids), we postulate that organic earthworm-excreted compounds, possibly amino acids and acetates, may have been involved. This would suggest a previously unknown shortcut in the terrestrial N and C cycle, however, it requires confirmation through advanced techniques such as compound-specific isotope analysis or liquid chromatography with mass spectrometry. These experimental results suggest that multitrophic linkages from living decomposers in soil to plants and aboveground consumers may happen at much shorter time scales than is generally assumed.

Leafminer attack accelerates the development of soil-dwelling conspecific pupae via plant-mediated changes in belowground volatiles.

Summary Herbivore population dynamics are strongly influenced by the interactions established through their shared host. Such plant‐mediated interactions can occur between different herbivore species and different life developmental stages of the same herbivore. However, whether these interactions occur between leaf‐feeding herbivores and their soil‐dwelling pupae is unknown.We studied whether tomato (Solanum lycopersicum) leaf herbivory by the American serpentine leafminer Liriomyza trifolii affects the performance of conspecific pupae exposed to the soil headspace of the plant. To gain mechanistic insights, we performed insect bioassays with the jasmonate‐deficient tomato mutant def‐1 and its wild‐type, along with phytohormones, gene expression and root volatiles analyses.Belowground volatiles accelerated leafminer metamorphosis when wild‐type plants were attacked aboveground by conspecifics. The opposite pattern was observed for def‐1 plants, in which aboveground herbivory slowed metamorphosis. Leafminer attack induced jasmonate and abscisic acid accumulation and modulated volatile production in tomato roots in a def‐1‐dependent manner.Our results demonstrate that aboveground herbivory triggers changes in root defence signalling and expression, which can directly or indirectly via changes in soil or microbial volatiles, alter pupal development time. This finding expands the repertoire of plant–herbivore interactions to herbivory‐induced modulation of metamorphosis, with potential consequences for plant and herbivore community dynamics.

Beneficial worm allies warn plants of parasite attack below-ground and reduce above-ground herbivore preference and performance.

Antagonistic interactions among different functional guilds of nematodes have been recognized for quite some time, but the underlying explanatory mechanisms are unclear. We investigated responses of tomato (Solanum lycopersicum) to two functional guilds of nematodes-plant parasite (Meloidogyne javanica) and entomopathogens (Heterorhabditis bacteriophora, Steinernema feltiae below-ground, and S.carpocapsae)-as well as a leaf mining insect (Tuta absoluta) above-ground. Our results indicate that entomopathogenic nematodes (EPNs): (1) reduced root knot nematode (RKN) infestation below-ground, (2) reduced herbivore (T.absoluta) host preference and performance above-ground, and (3) induced overlapping plant defence responses by rapidly activating polyphenol oxidase and guaiacol peroxidase activity in roots, but simultaneously suppressing this activity in above-ground tissues. Concurrently, we investigated potential plant signalling mechanisms underlying these interactions using transcriptome analyses. We found that both entomopathogens and plant parasites triggered immune responses in plant roots with shared gene expression. Secondary metabolite transcripts induced in response to the two nematode functional guilds were generally overlapping and showed an analogous profile of regulation. Likewise, we show that EPNs modulate plant defence against RKN invasion, in part, by suppressing active expression of antioxidant enzymes. Inoculations of roots with EPN triggered an immune response in tomato via upregulated phenylpropanoid metabolism and synthesis of protease inhibitors in plant tissues, which may explain decreased egg laying and developmental performance exhibited by herbivores on EPN-inoculated plants. Furthermore, changes induced in the volatile organic compound-related transcriptome indicated that M.javanica and/or S.carpocapsae inoculation of plants triggered both direct and indirect defences. Our results support the hypothesis that plants "mistake" subterranean EPNs for parasites, and these otherwise beneficial worms activate a battery of plant defences associated with systemic acquired resistance and/or induced systemic resistance with concomitant antagonistic effects on temporally co-occurring subterranean plant pathogenic nematodes and terrestrial herbivores.

The Influence of Above-Ground Herbivory on the Response of Arctic Soil Methanotrophs to Increasing CH4 Concentrations and Temperatures.

Rising temperatures in the Arctic affect soil microorganisms, herbivores, and peatland vegetation, thus directly and indirectly influencing microbial CH4 production. It is not currently known how methanotrophs in Arctic peat respond to combined changes in temperature, CH4 concentration, and vegetation. We studied methanotroph responses to temperature and CH4 concentration in peat exposed to herbivory and protected by exclosures. The methanotroph activity was assessed by CH4 oxidation rate measurements using peat soil microcosms and a pure culture of Methylobacter tundripaludum SV96, qPCR, and sequencing of pmoA transcripts. Elevated CH4 concentrations led to higher CH4 oxidation rates both in grazed and exclosed peat soils, but the strongest response was observed in grazed peat soils. Furthermore, the relative transcriptional activities of different methanotroph community members were affected by the CH4 concentrations. While transcriptional responses to low CH4 concentrations were more prevalent in grazed peat soils, responses to high CH4 concentrations were more prevalent in exclosed peat soils. We observed no significant methanotroph responses to increasing temperatures. We conclude that methanotroph communities in these peat soils respond to changes in the CH4 concentration depending on their previous exposure to grazing. This “conditioning” influences which strains will thrive and, therefore, determines the function of the methanotroph community.

Root infection by the nematode Meloidogyne incognita modulates leaf antiherbivore defenses and plant resistance to Spodoptera exigua.

Studies on plant-mediated interactions between root parasitic nematodes and aboveground herbivores are rapidly increasing. However, outcomes for the interacting organisms vary, and the mechanisms involved remain ambiguous. We hypothesized that the impact of root infection by the root-knot nematode Meloidogyne incognita on the performance of the aboveground caterpillar Spodoptera exigua is modulated by the nematode's infection cycle. We challenged root-knot nematode-infected tomato plants with caterpillars when the nematode's infection cycle was at the invasion, galling, and reproduction stages. We found that M. incognita root infection enhanced S. exigua performance during the galling stage, while it did not affect the caterpillar's performance at the invasion and reproduction stages. Molecular and chemical analyses performed at the different stages of the nematode infection cycle revealed that M. incognita root infection systemically affected the jasmonic acid-, salicylic acid-, and abscisic acid-related responses, as well as the changes in the leaf metabolome triggered during S. exigua feeding. The M. incognita-induced leaf responses varied over the nematode's root infection cycle. These findings suggest that specific leaf responses triggered systemically by the nematode at its different life-cycle stages underlie the differential impact of M. incognita on plant resistance against the caterpillar S. exigua.

Ecological impacts of the invasive plant Xanthium strumarium and the impacts of three aboveground herbivores on the invader

Exotic plant invasions are generally assumed to negatively impact biodiversity of recipient communities. However, experimental evidence for this assumption is little, especially in developing countries. In addition, there is hardly applicable method to control invasive plants. Enemy release is often hypothesized to facilitate exotic plant invasions, thus re-association with natural enemies of invasive ranges may inhibit invasions of exotic plants, and these enemies may have the potential to be used as biological control agents for these invaders. To address above problems, we determined abundance of the invasive plant Xanthium strumarium, its impacts on co-occurring native plants, and abundances of three herbivores and their impacts on the invader at five sites in the rice tract of Punjab, Pakistan. Relative abundance of the invader varied greatly among the study sites, ranging from 3.5% to 50.4%. The invader significantly reduced abundance of co-occurring native plants at all study sites, and their richness and evenness at some sites with high abundances of the invader. The polyphagous insects Phenacoccus solenopsis, Podisus maculiventris and Spodoptera frugiperda were all found on the invader (not on crops) at all study sites, which significantly reduced growth of the invader. The abundance of P. solenopsis on the invader was much higher than those of P. maculiventris and S. frugiperda. The abundances of the herbivores were significantly higher at sites with high relative to low relative abundances of the invader. Our study indicates that natural enemies may accumulate on invasive plants with the increase of their abundance and/or invasion time, inhibiting further invasions, and provides evidence for enemy release hypothesis. More studies are needed to determine whether these herbivores, especially P. solenopsis, could be used as biological control agents for the invader.

Aboveground herbivory causes belowground changes in twelve oak Quercus species: a phylogenetic analysis of root biomass and non‐structural carbohydrate storage

Plant ecosystem structure is understood to be a result of complex multitrophic interactions. Most multitrophic studies focus on plant aboveground adaptations to aboveground herbivore pressures, neglecting belowground adaptations in response to aboveground damage. Differential investment in root structures may allow plants to compensate for tissue loss or damage due to herbivores. Furthermore, phylogeny may constrain a plant's ability to adapt belowground. We examined the belowground responses of 12 species of oak Quercus to varying locations and intensities of simulated herbivore damage. We first established that oak belowground traits responded to aboveground herbivory by measuring patterns of investment in coarse versus fine root structures and re‐allocation of non‐structural carbohydrates (NSC) to root storage. We then tested whether phylogeny could explain variations in investment patterns using phylogenetic independent contrasts. Plant adaptations to aboveground herbivory included allocating biomass and carbon reserves to root structures, depending on the location and intensity of herbivore damage. NSC re‐allocation to root storage was observed when oak species experienced any type of damage, but damage to lateral tissues caused a greater re‐allocation than apical damage or control treatments. We found that most belowground responses to aboveground herbivory are species‐specific and may be adapted for environmental conditions or type of herbivory. Some responses to herbivore damage, such as changes in fine‐root mass and root sugar concentrations, were phylogenetically constrained. Phylogenetic constraints generally occur when there is severe damage at the apical meristem. Plants may adapt to aboveground tissue loss due to varying herbivore pressures (i.e. varying location and intensity of damage) by differentially investing in root types and NSC re‐allocation to root storage. Understanding linkages between and phylogenetic constraints of plant belowground responses to aboveground herbivory will improve our understanding of the ecological processes involved in multitrophic interactions.

Aboveground herbivory suppresses the arbuscular mycorrhizal symbiosis, reducing plant phosphorus uptake

Most terrestrial plants form associations with arbuscular mycorrhizal (AM) fungi, which are soil-dwelling microbial symbionts that provide plants with soil nutrients, while plants supply the fungi with carbon. The majority of these plants are also subject to herbivory from insects, thus tripartite interactions between insect herbivores, plants, and AM fungi are ubiquitous. This study assessed how aboveground herbivory from a generalist insect herbivore (Helicoverpa punctigera) affects the AM symbiosis in two C4 grass species (Bothriochloa macra and Dichanthium sericeum) and the consequences for host plant growth and nutrient uptake. Aboveground herbivory reduced root growth and carbon allocation belowground in both plant species, along with an associated reduction in arbuscular colonisation and phosphorus uptake. These findings suggest that, in accordance with the carbon-limitation hypothesis, herbivory can suppress the AM symbiosis by decreasing carbon belowground, potentially hindering AM fungal-enhanced nutrient acquisition from the soil.

Age-dependent resistance and tolerance of an invasive tree to above- and belowground herbivore interactions: Implications for invasiveness and biological control

Resistance (reducing herbivore attack) and tolerance (buffering the negative impacts of herbivory) of invasive plants to pressure imposed by aboveground (AG) or belowground (BG) herbivore interactions have been largely overlooked. Experimental manipulations of native and invasive populations consisting of plants of various ages can help elucidate the long-term adaptations of invasive plant genotypes. Here, we subjected first-year and second-year tallow trees (Triadica sebifera) from the native and the invasive range to AG herbivory by the specialist caterpillar Gadirtha fusca and/or BG herbivory by the root-knot nematode Meloidogyne incognita, and tested their interactive effects on levels of resistance or tolerance. The results showed that in comparison to native genotypes, the invasive tallow plants showed lower resistance (higher percent leaf damage) but higher tolerance (regrew faster) to AG damage from G. fusca herbivory alone, but showed similar resistance to nematode herbivory alone, regardless of plant age. Dissimilarly, only the younger (first-year) invasive plants showed higher tolerance to BG herbivory alone. For plants under combined attack by G. fusca and M. incognita, younger invasive plants showed a mixed defense strategy that had both higher resistance (fewer root-knots) and higher tolerance to the BG nematode, while the older (second-year) invasive tallow plants showed similar levels of resistance but greater AG tolerance to both AG and BG herbivores. Our results suggest that the efficacy of a biological agent might be affected by native resident generalist herbivores. Resistance and tolerance of invasive plants to AG-BG herbivory are likely to change as plants age. Higher tolerance of invasive plants like tallow to AG-BG herbivory in the area of introduction compared to the original range may explain the invasion success of the plants as well as the reduced efficacy of biocontrol agents.

Effects of soil biota on growth, resistance and tolerance to herbivory in Triadica sebifera plants

Interactions with soil microbes can strongly affect plant growth and defense against aboveground herbivores. Plant species often accumulate specific soil pathogens in their rhizosphere, leading to reduced growth of plants in soils originating from stands of conspecific plants compared to soils from heterospecific plants. However, whereas effects of such conspecific vs. heterospecific soil biota on plant growth have been well documented, their effects on plant resistance and tolerance to aboveground insect herbivores have not. We compared growth and defense of Triadica sebifera plants from populations where the species is native (China), when grown in sterilized soils, or in soils harbouring belowground biota from conspecific (native Triadica) or heterospecific (native grass) soils. In each of these soils, plants were exposed to a 15-day period of foliar herbivory by a specialist weevil (Heterapoderopsis bicallosicollis), a specialist caterpillar (Gadirtha inexacta), or no herbivory (cage), followed by a 60-day recovery period. Soil biota from conspecific and hetetospecific soils differed in their effects on plant growth and defense. First, in the absence of herbivory, soil biota from heterospecific soils slightly enhanced plant growth, whereas those from conspecific soils strongly reduced plant growth. Second, soil biota from conspecific soils strongly affected plant resistance and tolerance to foliar herbivory, whereas soil biota from heterospecific plants did not. The effects of soil biota on plant defense were herbivore-specific. In particular, conspecific soil biota reduced resistance to caterpillar but not to weevil feeding, whereas they enhanced tolerance to weevil but not to caterpillar feeding. Conspecific soil biota also mitigated induction of root flavonoids by herbivores and led to reduced root phenolics in response to herbivory. Conversely, caterpillar feeding increased AMF colonization, but under these conditions, AMF colonization was negatively associated with plant biomass. In addition to testing effects on native plants, we also tested effects of native soil biota on growth and resistance of plants from the introduced range (North America). Plants from the introduced range had higher shoot production, shoot-to-root ratio, and leaf phenolic and flavonoid production than plants from the native range, but their interactions with soil biota showed only minor differences compared to plants from the native range. Our results suggest that incorporating the effects of soil biota in interactions between plants and foliar herbivores is critical for understanding variations in growth, defense, and performance among plant populations at local and broader geographic scales.

While shoot herbivores reduce, root herbivores increase nutrient enrichment's impact on diversity in a grassland model.

Nutrient enrichment is widespread throughout grassland systems and expected to increase during the Anthropocene. Trophic interactions, like aboveground herbivory, have been shown to mitigate its effect on plant diversity. Belowground herbivory may also impact these habitats' response to nutrient enrichment, but its influence is much less understood, and likely to depend on factors such as the herbivores' preference for dominant species and the symmetry of belowground competition. If preferential toward the dominant, fastest growing species, root herbivores may reduce these species' relative fitness and support diversity during nutrient enrichment. However, as plant competition belowground is commonly considered to be symmetric, root herbivores may be less impactful than shoot herbivores because they do not reduce any competitive asymmetry between the dominant and subordinate plants. To better understand this system, we used an established, two-layer, grassland community model to run a full-factorially designed simulation experiment, crossing the complete removal of aboveground herbivores and belowground herbivores with nutrient enrichment. After 100yr of simulation, we analyzed communities' diversity, competition on the individual level, as well as their resistance and recovery. The model reproduced both observed general effects of nutrient enrichment in grasslands and the short-term trends of specific experiments. We found that belowground herbivores exacerbate the negative influence of nutrient enrichment on Shannon diversity within our model grasslands, while aboveground herbivores mitigate its effect. Indeed, data on individuals' above- and belowground resource uptake reveals that root herbivory reduces resource limitation belowground. As with nutrient enrichment, this shifts competition aboveground. Since shoot competition is asymmetric, with larger, taller individuals gathering disproportionate resources compared to their smaller, shorter counterparts, this shift promotes the exclusion of the smallest species. While increasing the root herbivores' preferences toward dominant species lessens their negative impact, at best they are only mildly advantageous, and they do very little reduce the negative consequences of nutrient enrichment. Because our model's belowground competition is symmetric, we hypothesize that root herbivores may be beneficial when root competition is asymmetric. Future research into belowground herbivory should account for the nature of competition belowground to better understand the herbivores' true influence.

Foliar Aphid Herbivory Alters the Tomato Rhizosphere Microbiome, but Initial Soil Community Determines the Legacy Effects

Aboveground herbivory can impact the root-associated microbiome, while simultaneously different soil microbial communities influence herbivore performance. It is currently unclear how these reciprocal top-down and bottom-up interactions between plants, insects and microbes vary across different soils and over successive plant generations. In this study, we examined top-down impacts of above-ground herbivory on the rhizosphere microbiome across different soils, assessed bottom-up impacts of soil microbial community variation on herbivore performance, and evaluated their respective contributions to soil legacy effects on herbivore performance. We used Macrosiphum euphorbiae (potato aphid) and Solanum pimpinellifolium (wild tomato) to capture pre-domestication microbiome interactions with a specialist pest. First, using 16S rRNA sequencing we compared bacterial communities associated with rhizospheres of aphid-infested and uninfested control plants grown in three different soils over three time points. High aphid infestation impacted rhizosphere bacterial diversity in a soil-dependent manner, ranging from a 22% decrease to a 21% increase relative to uninfested plants and explained 6–7% of community composition differences in two of three soils. We next investigated bottom-up and soil legacy effects of aphid herbivory by growing wild tomatoes in each of the three soils and a sterilized “no microbiome” soil, infesting with aphids (phase one), then planting a second generation (phase two) of plants in the soil conditioned with aphid-infested or uninfested control plants. In the first phase, aphid performance varied across plants grown in different soil sources, ranging from a 20 to 50% increase in aphid performance compared to the “no microbiome” control soil, demonstrating a bottom-up role for soil microbial community. In the second phase, initial soil community, but not previous aphid infestation, impacted aphid performance on plants. Thus, while herbivory altered the rhizosphere microbiome in a soil community-dependent manner, the bottom-up interaction between the microbial community and the plant, not top-down effects of prior herbivore infestation, affected herbivore performance in the following plant generation. These findings suggest that the bottom-up effects of the soil microbial community play an overriding role in herbivore performance in both current and future plant generations and thus are an important target for sustainable control of herbivory in agroecosystems.