Root Herbivory Research Articles

Root and shoot glucosinolate allocation patterns follow optimal defence allocation theory

Summary Optimal defence allocation theory (ODT) is one of the most prominent theoretical frameworks to explain the allocation of defence compounds within plants. It predicts that the most valuable and vulnerable plant organs have the highest levels of chemical defence. The ODT has been well worked out and experimentally tested for shoot defences, but not for root defences. To assess if ODT principles apply similarly to roots and shoots, we examined glucosinolates in above‐ground and below‐ground organs of nine plant species belonging to two families. In order to evaluate whether ODT equally applies to shoot and root organs, we designed a conceptual model in which above‐ground and below‐ground organs were assigned to orders of importance to plant performance. We hypothesized that organs constituting the plant's core structure are better protected than more distal organs. The nine plant species were cultivated, and their roots and shoots were harvested and divided into three orders for glucosinolate analysis. Using a specialist (Delia radicum) and a generalist (Amphimallon solstitiale) root herbivore, we also experimentally tested the hypothesis that the generalist herbivore prefers to feed on fine roots (FRs) with a low glucosinolate concentration, while the specialist prefers taproots (TRs) with a high glucosinolate concentration. We found that both in roots and shoots, the higher ordered core structural organs (TRs and stems) had the highest levels of glucosinolates. Below‐ground, TRs and lateral roots were better protected than the more distal, and less costly, FRs in seven out of nine species tested. The specialist root herbivore preferred feeding on the highly defended TRs, which is in line with what has been found for above‐ground specialist herbivores. Moreover, the glucosinolate concentration in roots overall was significantly higher than that in shoots. Synthesis. These results support the hypothesis that Optimal defence allocation theory (ODT) generally applies to glucosinolate allocation in above‐ground and below‐ground organs and may mainly serve to maintain the integrity of the main plant structure. Moreover, it suggests that above‐ground and below‐ground insect herbivores independently exert similar selection pressures on defence allocation patterns in roots and shoots.

Impacts of root herbivory on seedlings of three species of boreal forest trees

Current meta-analysis has identified the shortage of information on responses of woody plants to root damage as the most critical gap in studies of belowground insect herbivory. We explored the impacts of natural and simulated root herbivory on seedlings of three boreal forest-forming trees, Pinus sylvestris, Picea abies and Betula pendula. We compared the effects of natural herbivory and mechanical damage by placing one larva of the common cockchafer, Melolontha melolontha, of different instars (to manipulate root loss) into a pot with a one-year-old seedling or we excised a portion of the roots by inserting a 13-mm diameter cork borer into the soil. During the experiment, which lasted from 2 to 8 weeks, the larvae consumed 53–73% of root biomass, whereas the mechanical damage treatment removed 30–40% of roots. Seedling mortality was low (5%) across all treatments, but the aboveground biomass decreased by 3–43% due to natural herbivory and by 6–30% due to mechanical damage. The M. melolontha larvae consumed a larger amount of roots from B. pendula than from either P. sylvestris or P. abies, but the larvae showed similar relative growth rates and mass gains, indicating a different efficiency of conversion of ingested food (ECI). The values of ECI calculated for larvae of M. melolontha (2.3–3.5%) appeared much lower than the 17% reported earlier for larvae of foliage-feeding beetles, presumably due to the high energetic cost of locomotion in the soil. Consequently, root-feeding insects consume much more plant biomass than leaf-feeding insects do to gain similar body mass, and therefore they inflict relatively more damage to plants per unit of mass gained. Based on the biomass of insect herbivores in European forests, we estimate that they consume 0.5% of the available fine root biomass. The effects of low levels of root damage on growth and reproduction of woody plants remain virtually unknown, emphasising the need for studies of plant- and ecosystem-level effects of minor but chronic root herbivory.

Host plant colonisation by arbuscular mycorrhizal fungi stimulates immune function whereas high root silicon concentrations diminish growth in a soil-dwelling herbivore

Plant nutritional quality is dependent on soil nutrients and co-evolved soil microbial symbionts. Most plants associate with arbuscular mycorrhizal (AM) fungi, which alter their nutritional quality and silicon (Si) uptake from the soil. High Si concentrations reduce plant nutritional quality and can act as an effective defence both aboveground and belowground. The growth and immune function of insect herbivores is dependent on the quality of their host plants, hence the AM symbiosis and Si concentrations can impact insect growth and immunity via changes in host plant quality. The effects of AM fungi or Si on root herbivores are poorly quantified, while impacts on insect immunity are unknown. We investigated the effects of host plant colonisation by AM fungi and high root Si concentrations on plant quality alongside the growth of a root feeding insect and the immune response to entomopathogenic nematode infection.Two sugarcane varieties (Saccharum species hybrids L.) were grown under fully factorial treatment combinations of ± Si and AM/non-AM. Root feeding insects (Dermolepida albohirtum Waterhouse) fed on the plants and their immune function was assessed in a bioassay, while insect growth and root consumption were assessed in a feeding trial. We found high Si concentrations decreased insect growth and root consumption, the latter by 71%. Insect growth was reduced on plants associated with AM fungi, which was dependent on Si treatment and plant variety. Insect immunity increased by 62% on AM colonised plants, which negatively correlated with insect growth. These results demonstrate that the impacts of the AM symbiosis on root feeding insects can depend on Si availability and plant variety. Our study suggests that AM fungi can prime insect immunity, independent of host plant quality or Si concentrations, and the negative effects of AM fungi on soil dwelling insects involves immune function stimulation which, due to a growth-immunity trade-off, results in growth reduction.

In the presence of specialist root and shoot herbivory, invasive‐range Brassica nigra populations have stronger competitive effects than native‐range populations

SUMMARY The evolution of increased competitive ability (EICA) hypothesis predicts that release from specialist herbivores enables invasive plants to evolve increased growth. The most powerful tests of EICA hypothesis are provided by approaches that simultaneously assess the effects of specialist herbivory and competitive interactions. However, such approaches are extremely rare, and hence how simultaneous release from root and shoot herbivory influence competitive ability of invasive plants remains little understood. Here, we tested whether invasive‐range Brassica nigra plants have evolved increased competitive ability, and whether expression of competitive ability depends on separate and simultaneous effects of specialist root and shoot herbivory. To do this, we grew B. nigra plants from eight invasive‐range and eight native‐range populations in the presence vs. absence of competition with a community of native plant species, and in the absence vs. presence of separate and simultaneous damage by a specialist root herbivore (Delia radicum) and a specialist shoot herbivore (Plutella xylostella). Brassica nigra performance was assessed by measuring biomass production and flowering of individual B. nigra plants. In partial support of the EICA hypothesis, invasive‐range B. nigra had greater flowering than native‐range conspecifics in the absence of competition. However, contrary to a prediction of the EICA hypothesis, invasive‐range B. nigra produced less above‐ground biomass than native‐range B. nigra in the absence of shoot herbivory and competition. Moreover, with simultaneous root and shoot herbivory, invasive‐range B. nigra suppressed a competing community more strongly than native‐range B. nigra did. Synthesis. Our results suggest that invasiveness may be driven by mechanisms other than increased individual size. Simultaneous root and shoot herbivory in the invasive range may enhance suppressive effects of introduced plant species that have not completely escaped herbivore damage in the introduced range.

Tomato‐mediated interactions between root herbivores and aphids: insights into plant defence signalling

AbstractReactions of plants to insect pests include the activation of a local and systemic defence response. This response is based on transcriptional changes that are mainly controlled and coordinated by phytohormones. The above‐ and belowground part of plants can be challenged by different insects and therefore, the defence response to one attacker can influence other insects. The study of plant‐mediated interaction between pests that are physically or temporally separated yielded a variety of outcomes, with positive, negative, and neutral effects described in the literature. In this study, we examined possible plant‐mediated interactions between above‐ and belowground insect pests with different feeding guilds in tomato, Solanum lycopersicum L. (Solanaceae). Root feeding by Agriotes lineatus (L.) (Coleoptera: Elateridae) larvae caused a decrease in the development rate, fertility, and weight in Myzus persicae (Sulzer) (Hemiptera: Aphididae). To gain insights into the plant systemic signalling mechanisms, we also performed a time‐course expression analysis of defence‐related phytohormone marker genes. The result indicated a dynamic systemic response in leaves following root herbivory, which comprises the activation of genes dependent on different molecular pathways involved in plant stress response. Our work demonstrated that root herbivory increased aphid resistance in tomato and that a combination of signals enables the communication between below‐ and aboveground pests with different feeding guilds.

Impact of predicted precipitation scenarios on multitrophic interactions

Summary Predicted changes in the frequency and intensity of extreme rainfall events in the UK have the potential to disrupt terrestrial ecosystem function. However, responses of different trophic levels to these changes in rainfall patterns, and the underlying mechanisms, are not well‐characterised. This study aimed to investigate how changes in both the quantity and frequency of rainfall events will affect the outcome of interactions between plants, insect herbivores (above‐ and below‐ground) and natural enemies. Hordeum vulgare L. plants were grown in controlled conditions and in the field, and subjected to three precipitation scenarios: ambient (based on a local 10 year average rainfall); continuous drought (40% reduction compared with ambient); drought/deluge (40% reduction compared to ambient at a reduced frequency). The effects of these watering regimes and wireworm (Agriotes species) root herbivory on the performance of the plants, aphid herbivores above‐ground (Sitobion avenae, Metapolophium dirhodum, and Rhopalosiphum padi), and natural enemies of aphids including ladybirds (Harmonia axyridis) were assessed from measurements of plant growth, insect abundance and mass, and assays of feeding behaviour. Continuous drought decreased plant biomass, whereas reducing the frequency of watering events did not affect plant biomass but did alter plant chemical composition. In controlled conditions, continuous drought ameliorated the negative impact of wireworms on plant biomass. Compared with the ambient treatment, aphid mass was increased by 15% when feeding on plants subjected to drought/deluge; and ladybirds were 66% heavier when feeding on these aphids but this did not affect ladybird prey choice. In field conditions, wireworms feeding below‐ground reduced the number of shoot‐feeding aphids under ambient and continuous drought conditions but not under drought/deluge. Predicted changes in both the frequency and intensity of precipitation events under climate change have the potential to limit plant growth, but reduce wireworm herbivory, whilst simultaneously promoting above‐ground aphid numbers and mass, with these effects transferring to the third trophic level. Understanding the effect of future changes in the precipitation on species interactions is critical for determining their potential impact on ecosystem functioning and constructing accurate predictions under global change scenarios. A lay summary is available for this article.

Multitrophic interactions between maize mycorrhizas, the root feeding insect Phyllophaga vetula and the entomopathogenic fungus Beauveria bassiana

Root feeding larvae of Phyllophaga vetula causes severe damage in maize agroecosystems and their conventional management with chemical pesticides adversely affects the environment. Plant beneficial rhizosphere microorganisms such as mycorrhzial and entomopathogenic fungi (EPF) offer an alternative sustainable pest management strategy, but their successful integration in agroecosystems requires profound knowledge about their ecology. Here we explored interactions between a commercial isolate of the EPF Beauveria bassiana and indigenous maize mycorrhizas and their single and combined effects against root herbivory from the insect larvae P. vetula. Maize plants were grown in a greenhouse for seven weeks in pots with P limited soil with all possible combinations of the three main factors including native arbuscular mycorrhizal fungi (AMF) (without and with), EPF (without and with) and P. vetula (without and with).Root herbivory by P. vetula reduced plant growth and nutrition, and in addition reduced AMF root colonization. In contrast, AMF improved plant growth and P nutrition, but also reduced the population density of B. bassiana. Inoculation with B. bassiana increased root dry weight and counteracted a decrease in shoot N content from P. vetula root herbivory, despite that only 5% of P. vetula larvae were infected with B. bassiana. Dual inoculation with AMF and B. bassiana increased plant growth and N shoot content allowing the plants to compensate for the damage caused by P. vetula herbivory. In conclusion our results show that combination of native AMF populations and the EPF B. bassiana induce tolerance in maize from root herbivory by P. vetula in terms of plant growth performance and nutrition.

Invisible but consequential: root endophytic fungi have variable effects on belowground plant–insect interactions

AbstractEndophytic fungi are ubiquitous in nature and can play important roles in regulating plant–herbivore interactions. While some aboveground obligate symbionts are considered defensive mutualists of host plants, the importance of root endophytes in plant defense, especially against root‐feeding insects, remains unclear. This study aimed to investigate the effects of root fungal endophytes on plant resistance against belowground herbivores and the recovery of host plants from damage. We grew the common grassFestuca arundinacea(tall fescue) semi‐aeroponically in the laboratory and inoculated roots with one of five fungal endophytes isolated from field‐collected tall fescue or meadow soil. Endophyte‐inoculated and uninoculated control plants were subjected to feeding by larvae of the generalist root herbivoreRhizotrogus majalis(European chafer). Herbivory intensity was quantified after eight days, and regrowth of roots and shoots following root herbivory and mechanical shoot damage was measured thereafter for each treatment. Fungal identifications byDNAsequence analysis were conducted after completion of the herbivory experiments and revealed that the five endophytes included the decomposer fungiTrametes versicolorandMortierella alpina, the entomopathogenic fungiIsaria fumosoroseaandBeauveria bassiana, and a potential plant pathogen/entomopathogenFusariumcf.equiseti. The effects of these root endophytes on plant defense against root‐feeding insects were species‐specific. While four endophytes had few effects on plant resistance, endophyticB. bassianasignificantly reduced herbivore damage to roots. In comparison, plant tolerance to damage was impaired after colonization by all endophyte species exceptT. versicolorandM. alpina. The contrasting effects of endophytes on plant resistance and plant tolerance suggest that research solely evaluating plant resistance is likely to overestimate the benefits conferred by endophytes without accounting for potential negative effects on plant tolerance. We propose a conceptual framework to include both plant resistance and tolerance as two dimensions of a defensive strategy and show that plant associations with different root endophytes may shift the relative importance of resistance and tolerance for plant defense.

A Herbivore Tag-and-Trace System Reveals Contact- and Density-Dependent Repellence of a Root Toxin.

Foraging behavior of root feeding organisms strongly affects plant-environment-interactions and ecosystem processes. However, the impact of plant chemistry on root herbivore movement in the soil is poorly understood. Here, we apply a simple technique to trace the movement of soil-dwelling insects in their habitats without disturbing or restricting their interactions with host plants. We tagged the root feeding larvae of Melolontha melolontha with a copper ring and repeatedly located their position in relation to their preferred host plant, Taraxacum officinale, using a commercial metal detector. This method was validated and used to study the influence of the sesquiterpene lactone taraxinic acid β-D-glucopyranosyl ester (TA-G) on the foraging of M. melolontha. TA-G is stored in the latex of T. officinale and protects the roots from herbivory. Using behavioral arenas with TA-G deficient and control plants, we tested the impact of physical root access and plant distance on the effect of TA-G on M. melolontha. The larvae preferred TA-G deficient plants to control plants, but only when physical root contact was possible and the plants were separated by 5cm. Melolontha melolontha showed no preference for TA-G deficient plants when the plants were grown 15cm apart, which may indicate a trade-off between the cost of movement and the benefit of consuming less toxic food. We demonstrate that M. melolontha integrates host plant quality and distance into its foraging patterns and suggest that plant chemistry affects root herbivore behavior in a plant-density dependent manner.

Nematode Root Herbivory in Tomato Increases Leaf Defenses and Reduces Leaf Miner Oviposition and Performance

The outcome of plant-mediated interactions among herbivores from several feeding guilds has been studied intensively. However, our understanding on the effects of nematode root herbivory on leaf miner oviposition behavior and performance remain limited. In this study, we evaluated whether Meloidogyne incognita root herbivory affects Tuta absoluta oviposition preference on Solanum lycopersicum plants and the development of the resulting offspring. To investigate the M. incognita-herbivory induced plant systemic responses that might explain the observed biological effects, we measured photosynthetic rates, leaf trypsin protease inhibitor activities, and analyzed the profile of volatiles emitted by the leaves of root-infested and non-infested plants. We found that T. absoluta females avoided laying eggs on the leaves of root-infested plants, and that root infestation negatively affected the pupation process of T. absoluta. These effects were accompanied by a strong suppression of leaf volatile emissions, a decrease in photosynthetic rates, and an increase in the activity of leaf trypsin protease inhibitors. Our study reveals that root attack by nematodes can shape leaf physiology, and thereby increases plant resistance.

Get Tough, Get Toxic, or Get a Bodyguard: Identifying Candidate Traits Conferring Belowground Resistance to Herbivores in Grasses.

Grasses (Poaceae) are the fifth-largest plant family by species and their uses for crops, forage, fiber, and fuel make them the most economically important. In grasslands, which broadly-defined cover 40% of the Earth's terrestrial surface outside of Greenland and Antarctica, 40–60% of net primary productivity and 70–98% of invertebrate biomass occurs belowground, providing extensive scope for interactions between roots and rhizosphere invertebrates. Grasses invest 50–70% of fixed carbon into root construction, which suggests roots are high value tissues that should be defended from herbivores, but we know relatively little about such defenses. In this article, we identify candidate grass root defenses, including physical (tough) and chemical (toxic) resistance traits, together with indirect defenses involving recruitment of root herbivores' natural enemies. We draw on relevant literature to establish whether these defenses are present in grasses, and specifically in grass roots, and which herbivores of grasses are affected by these defenses. Physical defenses could include structural macro-molecules such as lignin, cellulose, suberin, and callose in addition to silica and calcium oxalate. Root hairs and rhizosheaths, a structural adaptation unique to grasses, might also play defensive roles. To date, only lignin and silica have been shown to negatively affect root herbivores. In terms of chemical resistance traits, nitrate, oxalic acid, terpenoids, alkaloids, amino acids, cyanogenic glycosides, benzoxazinoids, phenolics, and proteinase inhibitors have the potential to negatively affect grass root herbivores. Several good examples demonstrate the existence of indirect defenses in grass roots, including maize, which can recruit entomopathogenic nematodes (EPNs) via emission of (E)-β-caryophyllene, and similar defenses are likely to be common. In producing this review, we aimed to equip researchers with candidate root defenses for further research.

A whole-plant perspective reveals unexpected impacts of above- and belowground herbivores on plant growth and defense.

Trade-offs between plant growth and defense are central to theoretical frameworks used to study the ecology and evolution of plant defense against herbivores. However, these frameworks, as well as the experiments designed to test them, rarely include belowground herbivores. We experimentally challenged seedlings of the tropical shrub Solanum lycocarpum (Solanaceae) with either aboveground foliar herbivores (Spodoptera caterpillars) or belowground root herbivores (the nematode Meloidogyne incognita) and measured the resulting changes in plant growth rates, biomass allocation, and the concentration of defensive terpenoids in roots and leaves. We found that plants that suffered aboveground herbivory responded with aboveground growth but belowground defense. Similarly, belowground herbivory resulted in root growth but elevated defenses of leaves. These results underscore the importance of belowground plant-herbivore interactions, and suggest that, in contrast to theoretical predictions, plants can simultaneously invest in both growth and defense. Finally, they emphasize the need for a "whole-plant" perspective in theoretical and empirical evaluations of plant-herbivore interactions.

Increased root herbivory under elevated atmospheric carbon dioxide concentrations is reversed by silicon‐based plant defences

Summary Predicted increases in atmospheric concentrations of CO2 may alter the susceptibility of many plants to insect herbivores due to changes in plant nutrition and defences. Silicon plays a critical role in plant defence against herbivores, so increasing such silicon‐based defences in plants may help remediate situations where plants become more susceptible to herbivores. Sugar cane (Saccharum spp. hybrid) was subjected to fully factorial treatment combinations of ambient (aCO2) or elevated (eCO2) atmospheric CO2 concentrations; ambient silicon or silicon supplementation; insect‐free or subject to root herbivory by greyback canegrub (Dermolepida albohirtum). A glasshouse study was used to determine how these factors affected rates of photosynthesis, growth, chemistry (concentrations of silicon, carbon, nitrogen and non‐structural carbohydrates). Changes in canegrub mass were determined in the glasshouse pot study, together with more detailed assessment of how eCO2 and silicon supplementation affected performance and feeding behaviour (relative growth rate and relative consumption) in a 24‐h feeding efficiency assay. Elevated CO2 and silicon supplementation increased rates of photosynthesis (+32% and 14%, respectively) and sugar cane biomass (+45% and 69%, respectively). Silicon supplementation increased silicon concentrations in both leaves and roots by 54% and 75%, respectively. eCO2 caused root C : N to increase by 12%. Canegrub performance and consumption increased under eCO2; relative growth rate (RGR) increased by 116% and consumed 57% more root material (suggestive of compensatory feeding). Silicon application reversed these effects, with large decreases in mass change, RGR and root consumption (65% less root mass consumed). Synthesis and applications. Our results suggest future atmospheric carbon dioxide concentrations could lead to increased crop damage by a below‐ground herbivore. Increasing bioavailable silicon in soil stimulated silicon‐based defences which dramatically decreased herbivory and herbivore performance. Our findings suggest future pest management strategies could benefit from characterising deficiencies in bioavailable silicon in agricultural soils and targeted application of silicon fertilisers. Moreover, future breeding programmes should exploit variation in silicon uptake between cultivars to enhance silicon uptake in new crop varieties. Silicon‐based plant defence proved to be highly beneficial for remediating the negative effects of atmospheric change on sugar cane susceptibility to herbivory and could be applicable in other crops.

Erratum to: Belowground nitrogen transfer from legumes to non-legumes under managed herbaceous cropping systems. A review

Nitrogen is the most limiting nutrient in most agro-ecosystems and thus critical for sustaining high yields. Conventional agricultural practices use synthetic fertilizers to ensure an adequate supply of nitrogen in soils, but fertilizers come at a significant monetary and environmental cost. A strategy to improve nitrogen supply in cropping systems is the inclusion of nitrogen-fixing legumes, which can provide nitrogen benefits to companion crops through belowground nitrogen transfer. However, a better understanding of the underlying mechanisms and factors that govern nitrogen transfer is important in order to determine potential areas for improving this association. Here, we review the mechanisms of belowground nitrogen transfer in managed herbaceous cropping systems, focusing on forage systems. We classify three major routes of nitrogen transfer from legumes to non-legumes: (1) decomposition of legume root tissues and uptake of mineralized nitrogen by neighboring plants, (2) exudation of soluble nitrogen compounds by legumes and uptake by non-legumes, and (3) transfer of nitrogen mediated by plant-associated mycorrhizae. Literature data shows that rates of nitrogen transfer range from 0 to 73 % from forage legumes to companion grasses in mixed stands, depending on the legume species and cultivar. We list the factors that affect nitrogen transfer including abiotic factors, e.g., water stress, temperature, light, soil available nitrogen, and application of nitrogen fertilizer, and biotic factors, e.g., root contact, plant density, growth stage, production year, defoliation, and root herbivores. While the rates of nitrogen transfer are often constrained by abiotic conditions, such as temperature and water availability, that are beyond the control of growers, agronomic practices, e.g., planting density and choice of species and cultivar, may help to increase nitrogen transfer. Ultimately, the selection of plant pairs with compatible traits offers the best path forward to improving nitrogen transfer in intercrops.

Effects of population-related variation in plant primary and secondary metabolites on aboveground and belowground multitrophic interactions.

Insects feeding on aboveground and belowground tissues can influence each other through their shared plant and this is often mediated by changes in plant chemistry. We examined the effects of belowground root fly (Delia radicum) herbivory on the performance of an aboveground herbivore (Plutella xylostella) and its endoparasitoid wasp (Cotesia vestalis). Insects were reared on three populations of wild cabbage (Brassica oleracea) plants, exhibiting qualitative and quantitative differences in root and shoot defense chemistry, that had or had not been exposed to root herbivory. In addition, we measured primary (amino acids and sugars) and secondary [glucosinolate (GS)] chemistry in plants exposed to the various plant population-treatment combinations to determine to what extent plant chemistry could explain variation in insect performance variables using multivariate statistics. In general, insect performance was more strongly affected by plant population than by herbivory in the opposite compartment, suggesting that population-related differences in plant quality are larger than those induced by herbivory. Sugar profiles were similar in the three populations and concentrations only changed in damaged tissues. In addition to population-related differences, amino acid concentrations primarily changed locally in response to herbivory. Whether GS concentrations changed in response to herbivory (indole GS) or whether there were only population-related differences (aliphatic GS) depended on GS class. Poor correlations between performance and chemical attributes made biological interpretation of these results difficult. Moreover, trade-offs between life history traits suggest that factors other than food nutritional quality contribute to the expression of life history traits.

DRI-Grass: A New Experimental Platform for Addressing Grassland Ecosystem Responses to Future Precipitation Scenarios in South-East Australia.

Climate models predict shifts in the amount, frequency and seasonality of rainfall. Given close links between grassland productivity and rainfall, such changes are likely to have profound effects on the functioning of grassland ecosystems and modify species interactions. Here, we introduce a unique, new experimental platform – DRI-Grass (Drought and Root Herbivore Interactions in a Grassland) – that exposes a south-eastern Australian grassland to five rainfall regimes [Ambient (AMB), increased amount (IA, +50%), reduced amount (RA, -50%), reduced frequency (RF, single rainfall event every 21 days, with total amount unchanged) and summer drought (SD, 12–14 weeks without water, December–March)], and contrasting levels of root herbivory. Incorporation of a belowground herbivore (root-feeding scarabs) addition treatment allows novel investigation of ecological responses to the twin stresses of altered rainfall and root herbivory. We quantified effects of permanently installed rain shelters on microclimate by comparison with outside plots, identifying small shelter effects on air temperature (-0.19°C day, +0.26°C night), soil water content (SWC; -8%) and photosynthetically active radiation (PAR; -16%). Shelters were associated with modest increases in net primary productivity (NPP), particularly during the cool season. Rainfall treatments generated substantial differences in SWC, with the exception of IA; the latter is likely due to a combination of higher transpiration rates associated with greater plant biomass in IA and the low water-holding capacity of the well-drained, sandy soil. Growing season NPP was strongly reduced by SD, but did not respond to the other rainfall treatments. Addition of root herbivores did not affect plant biomass and there were no interactions between herbivory and rainfall treatments in the 1st year of study. Root herbivory did, however, induce foliar silicon-based defenses in Cynodon dactylon and Eragrostis curvula. Rapid recovery of NPP following resumption of watering in SD plots indicates high functional resilience at the site, and may reflect adaptation of the vegetation to historically high variability in rainfall, both within- and between years. DRI-Grass provides a unique platform for understanding how ecological interactions will be affected by changing rainfall regimes and, specifically, how belowground herbivory modifies grassland resistance and resilience to climate extremes.

Novel In vitro Procedures for Rearing a Root-Feeding Pest (Heteronychus arator) of Grasslands.

Optimizing plant protection against insect herbivory relies on testing plant defense mechanisms and how the insect response to these defensive strategies. Such experiments benefit from using insects generated from standardized rearing protocols since this reduces stochastic variation. Such protocols can be challenging to devise, however, especially for root herbivores. These insects generally have complex and long life cycles, which are often only poorly described. Moreover, using field-captured root herbivores is often suboptimal because it involves extensive excavation from sites selected by chance (their location is not obvious) and larval stages are frequently indistinguishable beyond the family level. We investigated in vitro procedures to improve the availability of the African Black Beetle (ABB) Heteronychus arator, an invasive alien pest in both Australia and New Zealand. Native to Africa, this scarab beetle has established in Australian and New Zealand grasslands, pastures, and crops. Adults feed on the stem of young plants just beneath the soil surface. During the mating season, gravid females lay eggs in the soil, giving rise to larvae feeding on grass roots, causing severe damage, and impairing plant growth. Here, we propose laboratory approaches to collect eggs from field-captured adult beetles, to hatch eggs, and to rear neonate larvae to adults. We propose that these methods will provide plant scientists and entomologists with a better and more controlled supply of ABB larvae for laboratory and field assays. In turn, this will assist with the collection of important information for the management of this insect pest and enhanced protection of plants in crop and grassland ecosystems.

Trade-Offs between Silicon and Phenolic Defenses may Explain Enhanced Performance of Root Herbivores on Phenolic-Rich Plants.

Phenolic compounds play a role in plant defense against herbivores. For some herbivorous insects, particularly root herbivores, host plants with high phenolic concentrations promote insect performance and tissue consumption. This positive relationship between some insects and phenolics, however, could reflect a negative correlation with other plant defenses acting against insects. Silicon is an important element for plant growth and defense, particularly in grasses, as many grass species take up large amounts of silicon. Negative impact of a high silicon diet on insect herbivore performance has been reported aboveground, but is unreported for belowground herbivores. It has been hypothesized that some silicon accumulating plants exhibit a trade-off between carbon-based defense compounds, such as phenolics, and silicon-based defenses. Here, we investigated the impact of silicon concentrations and total phenolic concentrations in sugarcane roots on the performance of the root-feeding greyback canegrub (Dermolepida albohirtum). Canegrub performance was positively correlated with root phenolics, but negatively correlated with root silicon. We found a negative relationship in the roots between total phenolics and silicon concentrations. This suggests the positive impact of phenolic compounds on some insects may be the effect of lower concentrations of silicon compounds in plant tissue. This is the first demonstration of plant silicon negatively affecting a belowground herbivore.

Trade‐offs and tritrophic consequences of host shifts in specialized root herbivores

Summary Trade‐offs in an herbivore's ability to feed, avoid predation and succeed on alternative hosts are thought to be major driving factors in host specialization. In this study, we compared how two closely related milkweed beetles (Tetraopes spp.) that have specialized on separate Asclepias species respond to host switching to alternative milkweed plants. By additionally examining effects on the beetles’ entomopathogenic natural enemies, we test whether host plant specialization is driven by plant–herbivore interactions alone or whether there is a role for the tritrophic interactions. Tetraopes beetles feed on milkweed roots as larvae and on milkweed shoots as adults, and they sequester toxic cardenolides from their host plants. We predicted that growth and sequestration would be compromised on alternative hosts as a consequence of specialization. Larvae of each Tetraopes species were reared on their native host plant, the host plant of the other species and two closely related Asclepias species that do not typically host Tetraopes. Regardless of beetle specialization, survival of both beetles was highest on A. syriaca, which has the second‐to‐lowest levels of root cardenolides and is the native host for T. tetraophthalmus. Larval growth was not affected by host plant for T. texanus, but T. tetraophthalmus grew best on A. exaltata, a close relative of A. syriaca that does not typically experience beetle feeding. We found that larval sequestration of cardenolides largely mirrored host plant values in the roots, with the exception of T. texanus on A. asperula, which sequestered a low proportion of the cardenolides from this host. Although all larvae were susceptible to entomopathogenic nematodes (EPNs), the number of EPNs emerging from cadavers depended on the beetle–plant combination, with more EPNs produced when beetles were feeding on native compared to non‐native hosts. The observed consequences for tritrophic interactions did not fit expectations for specialized herbivores (i.e. EPN predation was highest when beetles were on their native hosts), suggesting that specialization may be driven primarily by a plant–herbivore interaction in this system, not by a tritrophic effect. A Lay Summary is available for this article.

Root Herbivores Drive Changes to Plant Primary Chemistry, but Root Loss Is Mitigated under Elevated Atmospheric CO2.

Above- and belowground herbivory represents a major challenge to crop productivity and sustainable agriculture worldwide. How this threat from multiple herbivore pests will change under anthropogenic climate change, via altered trophic interactions and plant response traits, is key to understanding future crop resistance to herbivory. In this study, we hypothesized that atmospheric carbon enrichment would increase the amount (biomass) and quality (C:N ratio) of crop plant resources for above- and belowground herbivore species. In a controlled environment facility, we conducted a microcosm experiment using the large raspberry aphid (Amphorophora idaei), the root feeding larvae of the vine weevil (Otiorhynchus sulcatus), and the raspberry (Rubus idaeus) host-plant. There were four herbivore treatments (control, aphid only, weevil only and a combination of both herbivores) and an ambient (aCO2) or elevated (eCO2) CO2 treatment (390 versus 650 ± 50 μmol/mol) assigned to two raspberry cultivars (cv Glen Ample or Glen Clova) varying in resistance to aphid herbivory. Contrary to our predictions, eCO2 did not increase crop biomass or the C:N ratio of the plant tissues, nor affect herbivore abundance either directly or via the host-plant. Root herbivory reduced belowground crop biomass under aCO2 but not eCO2, suggesting that crops could tolerate attack in a CO2 enriched environment. Root herbivory also increased the C:N ratio in leaf tissue at eCO2, potentially due to decreased N uptake indicated by lower N concentrations found in the roots. Root herbivory greatly increased root C concentrations under both CO2 treatments. Our findings confirm that responses of crop biomass and biochemistry to climate change need examining within the context of herbivory, as biotic interactions appear as important as direct effects of eCO2 on crop productivity.