Medicago truncatula mutants demonstrate the role of plant calcium oxalate crystals as an effective defense against chewing insects

Calcium oxalate is the most abundant insoluble mineral found in plants and its crystals have been reported in more than 200 plant families. In the barrel medic Medicago truncatula Gaertn., these crystals accumulate predominantly in a sheath surrounding secondary veins of leaves. Mutants of M. truncatula with decreased levels of calcium oxalate crystals were used to assess the defensive role of this mineral against insects. Caterpillar larvae of the beet armyworm Spodoptera exigua Hübner show a clear feeding preference for tissue from calcium oxalate-defective (cod) mutant lines cod5 and cod6 in choice test comparisons with wild-type M. truncatula. Compared to their performance on mutant lines, larvae feeding on wild-type plants with abundant calcium oxalate crystals suffer significantly reduced growth and increased mortality. Induction of wound-responsive genes appears to be normal in cod5 and cod6, indicating that these lines are not deficient in induced insect defenses. Electron micrographs of insect mouthparts indicate that the prismatic crystals in M. truncatula leaves act as physical abrasives during feeding. Food utilization measurements show that, after consumption, calcium oxalate also interferes with the conversion of plant material into insect biomass during digestion. In contrast to their detrimental effects on a chewing insect, calcium oxalate crystals do not negatively affect the performance of the pea aphid Acyrthosiphon pisum Harris, a sap-feeding insect with piercing-sucking mouthparts. The results confirm a long-held hypothesis for the defensive function of these crystals and point to the potential value of genes controlling crystal formation and localization in crop plants.     

An Assessment of Engineered Calcium Oxalate Crystal Formation on Plant Growth and Development as a Step toward Evaluating Its Use to Enhance Plant Defense

The establishment of new approaches to control chewing insects has been sought not only for direct use in reducing crop loss but also in managing resistance to the pesticides already in use. Engineered formation of calcium oxalate crystals is a potential strategy that could be developed to fulfill both these needs. As a step toward this development, this study investigates the effects of transforming a non-calcium oxalate crystal accumulating plant, Arabidopsis thaliana, into a crystal accumulating plant. Calcium oxalate crystal accumulating A. thaliana lines were generated by ectopic expression of a single bacterial gene encoding an oxalic acid biosynthetic enzyme. Biochemical and cellular studies suggested that the engineered A. thaliana lines formed crystals of calcium oxalate in a manner similar to naturally occurring crystal accumulating plants. The amount of calcium oxalate accumulated in leaves also reached levels similar to those measured in the leaves of Medicago truncatula in which the crystals are known to play a defensive role. Visual inspection of the different engineered lines, however, suggested a phenotypic consequence on plant growth and development with higher calcium oxalate concentrations. The restoration of a near wild-type plant phenotype through an enzymatic reduction of tissue oxalate supported this observation. Overall, this study is a first to provide initial insight into the potential consequences of engineering calcium oxalate crystal formation in non-crystal accumulating plants.     

Increased calcium bioavailability in mice fed genetically engineered plants lacking calcium oxalate

Bioavailable calcium affects bone formation and calcification. Here we investigate how a single gene mutation altering calcium partitioning in the model forage crop Medicago truncatula affects calcium bioavailability. Previously, the cod5 M. truncatula mutant was identified which contains identical calcium concentrations to wild-type, but contains no oxalate crystals. In this study, equal number of male and female mice were randomly grouped and then fed one of four ⁴⁵Ca-containing diets: M. truncatula extrinsically or intrinsically labeled, and cod5 extrinsically or intrinsically labeled. Absorption of the tracer was determined in the legs one day after consumption. The absorption was similar in the M. truncatula and cod5 extrinsically labeled diets; however, in the intrinsically labeled diets, calcium absorption was 22.87% (P < 0.001) higher in mice fed cod5. Our study presents the first genetic evidence demonstrating the nutritional impact of removing oxalate crystals from foods.     

High levels of arbuscular mycorrhizal fungus colonization on Medicago truncatula reduces plant suitability as a host for pea aphids (Acyrthosiphon pisum)

This study sheds light on a poorly understood area in insect-plant-microbe interactions,focusing on aphid probing and feeding behavior on plants with varying levels of arbuscular mycorrhizal(AM)fungus root colonization.It investigates a commonly occurring interaction of three species:pea aphid Acyrthosiphon pisum,barrel medic Medicago truncatula,and the AM fungus Rhizophagus irregularis,examining whether aphid-feeding behavior changes when insects feed on plants at different levels of AM fungus colonization(42% and 84% root length colonized).Aphid probing and feeding behavior was monitored throughout 8 h of recording using the electrical penetration graph(EPG)technique,also,foliar nutrient content and plant growth were measured.Summarizing,aphids took longer to reach their 1st sustained phloem ingestion on the 84% AM plants than on the 42% AM plants or on controls.Less aphids showed phloem ingestion on the 84% AM plants relative to the 42% AM plants.Shoots of the 84% AM plants had higher percent carbon(43.7%)relative to controls(40.5%),and the 84% AM plants had reduced percent nitrogen(5.3%)relative to the 42% AM plants(6%).In conclusion,EPG and foliar nutrient data support the hypothesis that modifications in plant anatomy(e.g.,thicker leaves),and poor food quality(reduced nitrogen)in the 84% AM plants contribute to reduced aphid success in locating phloem and ultimately to differences in phloem sap ingestion.This work suggests that M.truncatula plants benefit from AM symbiosis not only because of increased nutrient uptake but also because of reduced susceptibility to aphids.     

Influence of presence and spatial arrangement of belowground insects on host‐plant selection of aboveground insects: a field study

Abstract 1. Several studies have shown that above‐ and belowground insects can interact by influencing each others growth, development, and survival when they feed on the same host‐plant. In natural systems, however, insects can make choices on which plants to oviposit and feed. A field experiment was carried out to determine if root‐feeding insects can influence feeding and oviposition preferences and decisions of naturally colonising foliar‐feeding insects. 2. Using the wild cruciferous plant Brassica nigra and larvae of the cabbage root fly Delia radicum as the belowground root‐feeding insect, naturally colonising populations of foliar‐feeding insects were monitored over the course of a summer season. 3. Groups of root‐infested and root‐uninfested B. nigra plants were placed in a meadow during June, July, and August of 2006 for periods of 3 days. The root‐infested and the root‐uninfested plants were either dispersed evenly or placed in clusters. Once daily, all leaves of each plant were carefully inspected and insects were removed and collected for identification. 4. The flea beetles Phyllotreta spp. and the aphid Brevicoryne brassicae were significantly more abundant on root‐uninfested (control) than on root‐infested plants. However, for B. brassicae this was only apparent when the plants were placed in clusters. Host‐plant selection by the generalist aphid M. persicae and oviposition preference by the specialist butterfly P. rapae, however, were not significantly influenced by root herbivory. 5. The results of this study show that the presence of root‐feeding insects can affect feeding and oviposition preferences of foliar‐feeding insects, even under natural conditions where many other interactions occur simultaneously. The results suggest that root‐feeding insects play a role in the structuring of aboveground communities of insects, but these effects depend on the insect species as well as on the spatial distribution of the root‐feeding insects.     

Entrapped sand as a plant defence: effects on herbivore performance and preference

1. Abrasive material in the diet of herbivorous organisms comes from a variety of sources, including crystalline silica or calcium in plant tissues, accidentally ingested soil while digging or grazing, and entrapped substrate on the surfaces of plants. A wide variety of plants entrap substrate, usually with glandular trichomes. 2. A previous study demonstrated that entrapped sand provided resistance to herbivory in the field. In this study, the following questions were addressed: how does entrapped sand on Abronia latifolia (Nyctaginaceae) leaves and stems affect preference and performance of a common herbivore, the large‐bodied caterpillar Hyles lineata (Sphingidae); does this effect differ from those experienced by an internally feeding leaf miner? 3. Using a combination of experimental and observational approaches, it was found that sand comprised ～4–5% of ingested weight during normal feeding of H. lineata caterpillars. This entrapped sand caused extensive wear to their mandibles, they avoided sand‐covered plants when given the choice, and the sand negatively impacted performance metrics, including pupal weight, development time, and growth rate. In contrast, a leaf‐mining caterpillar did not have a preference for or against feeding on sandy plants. 4. These results are similar to studies on mandibular wear due to grasses, and herbivorous insects that feed on these two plant groups may have similar morphologies. It is hypothesised that increased wear potential may be a convergent solution to abrasive plants in both mammals (hypsodonty) and insects.     

Interactive direct and plant‐mediated effects of elevated atmospheric [CO2] and temperature on a eucalypt‐feeding insect herbivore

Understanding the direct and indirect effects of elevated [CO₂] and temperature on insect herbivores and how these factors interact are essential to predict ecosystem‐level responses to climate change scenarios. In three concurrent glasshouse experiments, we measured both the individual and interactive effects of elevated [CO₂] and temperature on foliar quality. We also assessed the interactions between their direct and plant‐mediated effects on the development of an insect herbivore of eucalypts. Eucalyptus tereticornis saplings were grown at ambient or elevated [CO₂] (400 and 650 μmol mol^?1 respectively) and ambient or elevated ( + 4 °C) temperature for 10 months. Doratifera quadriguttata (Lepidoptera: Limacodidae) larvae were feeding directly on these trees, on their excised leaves in a separate glasshouse, or on excised field‐grown leaves within the temperature and [CO₂] controlled glasshouse. To allow insect gender to be determined and to ensure that any sex‐specific developmental differences could be distinguished from treatment effects, insect development time and consumption were measured from egg hatch to pupation. No direct [CO₂] effects on insects were observed. Elevated temperature accelerated larval development, but did not affect leaf consumption. Elevated [CO₂] and temperature independently reduced foliar quality, slowing larval development and increasing consumption. Simultaneously increasing both [CO₂] and temperature reduced these shifts in foliar quality, and negative effects on larval performance were subsequently ameliorated. Negative nutritional effects of elevated [CO₂] and temperature were also independently outweighed by the direct positive effect of elevated temperature on larvae. Rising [CO₂] and temperature are thus predicted to have interactive effects on foliar quality that affect eucalypt‐feeding insects. However, the ecological consequences of these interactions will depend on the magnitude of concurrent temperature rise and its direct effects on insect physiology and feeding behaviour.     

Cry1Ab protein levels in phytophagous insects feeding on transgenic corn: implications for secondary exposure risk assessment

Enzyme-linked immunosorbent assays (ELISA) and bioassays were used to estimate levels of Cry1Ab protein in four species of phytophagous insects after feeding on transgenic Bt-corn plants expressing Cry1Ab protein or artificial diets containing Cry1Ab protein. The level of Cry1Ab in insects feeding on sources containing the Cry1Ab protein was uniformly low but varied with insect species as well as food source. For the corn leaf aphid, Rhopalosiphum maidis (Fitch), feeding on diet solutions containing Cry1Ab protein, the level of the protein in the aphid was 250–500 times less than the original levels in the diet, whereas no Cry1Ab was detected by ELISA in aphids feeding on transgenic Bt-Corn plants. For the lepidopteran insects, Ostrinia nubilalis (Hübner), Helicoverpa zea (Boddie), and Agrotis ipsilon (Hufnagel), levels of Cry1Ab in larvae varied significantly with feeding treatment. When feeding for 24 h on artificial diets containing 20 and 100 ppm of Cry1Ab, the level of Cry1Ab in the larvae was about 57 and 142 times lower, respectively, than the original protein level in the diet for O. nubilalis, 20 and 34 times lower for H. zea, and 10 to 14 times lower for A. ipsilon. Diet incorporation bioassays with a susceptible insect (first instar O. nubilalis) showed significant Cry1Ab bioactivity present within whole body tissues of R. maidis and O. nubilalis that had fed on diet containing a minimum of 20 ppm or higher concentrations (100 or 200 ppm) of Cry1Ab, but no significant bioactivity within the tissues of these insects after feeding on transgenic Bt-corn plants. The relevance of these findings to secondary exposure risk assessment for transgenic Bt crops is discussed.     

Contrasting calcium localization and speciation in leaves of the Medicago truncatula mutant cod5 analyzed via synchrotron X–ray techniques

Oxalate‐producing plants accumulate calcium oxalate crystals (CaOx_(c)) in the range of 3–80% w/w of their dry weight, reducing calcium (Ca) bioavailability. The calcium oxalate deficient 5 (cod5) mutant of Medicago truncatula has been previously shown to contain similar Ca concentrations to wild‐type (WT) plants, but lower oxalate and CaOx_(c) concentrations. We imaged the Ca distribution in WT and cod5 leaflets via synchrotron X–ray fluorescence mapping (SXRF). We observed a difference in the Ca distribution between cod5 and WT leaflets, manifested as an abundance of Ca in the interveinal areas and a lack of Ca along the secondary veins in cod5, i.e. the opposite of what is observed in WT. X–ray microdiffraction (μXRD) of M. truncatula leaves confirmed that crystalline CaOx_(c) (whewellite; CaC₂O₄·H₂O) was present in the WT only, in cells sheathing the secondary veins. Together with μXRD, microbeam Ca K–edge X–ray absorption near‐edge structure spectroscopy (μXANES) indicated that, among the forms of CaOx, i.e. crystalline or amorphous, only amorphous CaOx was present in cod5. These results demonstrate that deletion of COD5 changes both Ca localization and the form of CaOx within leaflets.     

Finish line plant–insect interactions mediated by insect feeding mode and plant interference: a case study of Brassica interactions with diamondback moth and turnip aphid

There are gaps in our understanding of plant responses under different insect phytophagy modes and their subsequent effects on the insect herbivores’ performance at late season. Here we compared different types of insect feeding by an aphid, Lipaphis erysimi, and a lepidopteran, Plutella xylostella, and how this affected defensive metabolites in leaves of 2 Brassica species when plants gain maturity. Thiocyanate concentrations after P. xylostella and L. erysimi feeding activities were the same. Total phenolics was higher after the phloem feeder feeding than the folivore activity. The plants compensatory responses (i.e., tolerance) to L. erysimi feeding was significantly higher than the responses to P. xylostella. This study showed that L. erysimi had higher carbon than P. xylostella whereas nitrogen in P. xylostella was 1.42 times that in L. erysimi. Population size of the phloem feeder was not affected by plant species or insect coexistence. However, there was no correlation between plant defensive metabolites and both insects’ population size and biomass. This suggests that plant root biomass and tolerance index after different insect herbivory modes are not necessarily unidirectional. Importantly, the interaction between the folivore and the phloem feeder insects is asymmetric and the phloem feeder might be a trickier problem for plants than the folivore. Moreover, as both plants’ common and special defenses decreased under interspecific interference, we suggest that specialist insect herbivores can be more challenged in ecosystems in which plants are not involved in interspecific interference.     

Root damage to apple plants by cockchafer larvae induces a change in volatile signals below‐ and above‐ground

Volatile organic compounds (VOCs) mediate communication between plants and insects. Plants under insect herbivore attack release VOCs either at the site of attack or systemically, indicating within‐plant communication. Some of these VOCs, which may be induced only upon herbivore attack, recruit parasitoids and predatory insects to feed on the attacking insects. Moreover, some plants are able to ‘eavesdrop’ on herbivore‐induced plant volatiles (HIPVs) to prime themselves against impending attack; such eavesdropping exemplifies plant–plant communication. In apple orchards, the beetle Melolontha melolontha L. (Coleoptera: Scarabaeidae) is an important insect pest whose larvae live and feed on roots for about 4 years. In this study, we investigated whether the feeding activity of M. melolontha larvae (1) alters the volatile profile of apple roots, (2) induces the release of HIPVs systemically in the leaves, and (3) whether infested plants communicate to neighbouring non‐infested conspecifics through HIPVs. To answer these questions, we collected constitutive VOCs from intact M9 roots as well as M. melolontha larvae‐damaged roots using a newly designed ‘rhizobox’, to collect root‐released volatiles in situ, without damaging the plant root system. We also collected VOCs from the leaf‐bearing shoots of M9 whose roots were under attack by M. melolontha larvae and from shoots of neighbouring non‐infested conspecifics. Gas chromatography‐mass spectrometry analysis showed that feeding activity of M. melolontha larvae induces the release of specific HIPVs; for instance, camphor was found in the roots only after larvae caused root damage. Melolontha melolontha also induced the systemic release of methyl salicylate and (E,E)‐α‐farnesene from the leaf‐bearing shoots. Methyl salicylate and (E,E)‐α‐farnesene were also released by the shoots of non‐infested neighbouring conspecifics. These phenomena indicate the induction of specific VOCs below‐ and above‐ground upon M. melolontha larvae feeding on apple roots as well as plant–plant communication in apple plants.     

堇菜叶片草酸钙晶体与水分维持的关系

随着全球气候变化加重,干旱强度和持续时间逐渐增加,严重影响植物生长和作物产量。喀斯特为典型的干旱和高钙生境,植物叶片富集大量的草酸钙晶体,而该晶体与植物耐旱性之间的关系并不清楚。该研究以喀斯特适生植物堇菜(Viola verecumda)为材料,土壤进行自然干旱,分析堇菜叶片的草酸钙晶体变化特征与水分之间的关系。结果表明:在土壤自然干旱条件下,堇菜主要通过细胞内束缚水的释放,维持细胞内水分平衡;而在干旱后期,叶片通过关闭气孔,将部分自由水转变为束缚水,防止水分流失。此外,草酸钙晶体的密度与束缚水含量具有极其显著的强正相关线性回归关系(r=0.825 3,P0.000 1),表明草酸钙晶体作为主要的束缚水物质。因此,堇菜植物在耐旱过程中可能协调草酸钙晶体和气孔的生理行为忍耐干旱胁迫。     

Do defoliating insects distinguish between symmetric and asymmetric leaves within a plant?

1. Plants represent a highly heterogeneous resource for herbivores. One dimension of this heterogeneity is reflected by the within‐plant variation in the leaf fluctuating asymmetry (FA), i.e. in the magnitude of the random deviations from the symmetry in leaf shape. 2. This study is the first to test experimentally the hypothesis that variation in the quality of individual leaves for defoliating insects (11 species) within a plant (seven tree and shrub species) is associated with the FA of these leaves. 3. It was demonstrated that specialist defoliators generally distinguish between nearly symmetric (low FA) and highly asymmetric (high FA) leaves, but do not distinguish between discs cut from these leaves. Low‐FA leaves of Salix caprea, Salix myrsinifolia and Populus tremula were of better quality for insects than high‐FA leaves, as demonstrated by both preference tests and performance trials. By contrast, high‐FA leaves of Betula pubescens were of better quality for insects than low‐FA leaves, whereas insects feeding on Alnus incana showed similar responses to high‐ and low‐FA leaves. 4. It is concluded that insect herbivores can distinguish between leaves with high and low FA, and that FA may be associated with the quality of an individual leaf for insects, although the direction and strength of the effect of leaf FA on insect preference and performance vary among study systems. The ecological significance of substantial within‐plant variation in leaf FA remains to be explored.     

Comparative feeding selectivity of herbivorous insects on water lilies: aquatic vs. semi‐terrestrial insects and submersed vs. floating leaves

1. The rate of grazing damage experienced by submersed and floating leaves of water lilies (Nuphar variegata and Nymphaea odorata) was monitored in lakes in the Upper Peninsula of Michigan, U.S.A. Herbivores damaged 0.2–1.7% of the leaf surface of water lilies per day. These grazing rates differed between plant species, between submersed and floating leaves, and between lakes. Some leaves had more than 60% of their surface damaged and an overall mean of 16% damage occurred during the 2–3 week monitoring period of this study. 2. Snapshot measurements of grazing damage on randomly collected submersed and floating leaves of Nuphar showed that submersed leaves were more damaged (11.0 ± 1.6%, n = 84) than floating leaves (3.8 ± 0.6%, n = 92). Overall, these 176 Nuphar leaves had 7.2% of their area damaged. 3. Five species of herbivorous insects were commonly found on water lilies (Nymphaeacea). One primarily aquatic insect (sensu 1 ), a caddisfly larva (Trichoptera: Limniphilidae), had a generalized diet of water lilies, other macrophytes, algae, and detritus. Four of the five insects were from primarily terrestrial insect groups (Coleoptera and Diptera;‘secondary invaders’, sensu 1 ) and consumed only water lilies in food preference experiments. 4. The feeding preferences of the generalist trichopteran were altered when the macrophytes were freeze-dried, ground into a powder, and reconstituted in an alginate gel. This suggests that plant structure may be an important feeding determinant for this insect. In contrast, a specialist weevil preferred its host plant in choice assays, regardless of whether fresh tissue or reconstituted macrophytes were used, suggesting this insect cued on a unique, non-structural property of its host plant. 5. These results suggest that herbivory on freshwater macrophytes is of a similar magnitude to that on terrestrial plants. The findings of this study are consistent with the hypothesis that herbivorous insects of primarily terrestrial groups have a narrower diet breadth than insects of primarily aquatic groups.     

Feeding preferences of Melanoplus femurrubrum grasshoppers on native and exotic grasses: behavioral and molecular approaches

Generalist insect herbivores, such as grasshoppers, may either avoid feeding on exotic plants, potentially enabling these plants to become invasive in the introduced range, or insects may incorporate exotic plants into their diet, contributing to the biotic resistance of native communities and potentially preventing plant invasions. Accurate determination of insect diet preferences with regard to native and exotic plants can be challenging, but this information is critical for understanding the interaction between native herbivores and exotic plants, and ultimately the mechanisms underlying plant invasions. To address this, we combined behavioral and molecular approaches to accurately compare food consumption of the polyphagous red‐legged grasshopper, Melanoplus femurrubrum (De Geer) (Orthoptera: Acrididae), on native [Andropogon gerardii Vitman and Bouteloua curtipendula (Michx.) Torr.] and exotic, potentially invasive grasses [Miscanthus sinensis Andersson and Bothriochloa ischaemum (L.) Keng] (all Poaceae). We found that M. femurrubrum grasshoppers demonstrated strong feeding preferences toward exotic grasses in experiments with intact plants under both field and greenhouse conditions, but they showed no preference in experiments with clipped leaves. Additionally, we sampled the gut contents of M. femurrubrum collected in the field and identified the ingested plant species based on DNA sequences for the non‐coding region of the chloroplast trnL (UAA) gene. We found that exotic plants were prevalent in the gut contents of grasshoppers collected at study sites in Ohio and Maryland, USA. These results suggest that the generalist herbivore M. femurrubrum does not avoid feeding on exotic grasses with which they do not share coevolutionary history. In addition, by demonstrating greater food consumption of exotic plants, these grasshoppers potentially provide biotic resistance should these grasses escape cultivation and become invasive in the introduced range.     

Florivores on the dioecious shrub Eurya japonica and the preferences and performances of two polyphagous geometrid moths on male and female plants

Flowers of dioecious plants have sexually dimorphic traits that may affect florivore performances, and florivores may have preferences to plant sex that are correlated with their performance on different plant sexes. We investigated the florivory on a polygamodioecious evergreen shrub Eurya japonica in Japan to reveal florivores and their feeding patterns involved in sexually biased florivory on E. japonica flowers. Flowers of E. japonica were infested by lepidopteran and dipteran larvae and hemipteran insects. Lepidopteran larvae were chewers, dipteran larvae were gall makers and hemipteran insects were suckers. Chewed flowers were most frequent among infested flowers. Of florivores, lepidopteran larvae, mostly of Geometridae, were the important florivore that damaged flowers by chewing. Florivores infested male flower buds more often than female flower buds, but only a geometrid larvae Chloroclystis excise, which exclusively uses flower buds, showed the biased infection on male flowers. Rearing experiments for two other geometrid moths which use both leaves and flowers showed that the preference and performance of Ourapteryx nivea that fed mainly leaves did not differ between the plant sexes, whereas the development of Alcis angulifera larvae which fed both leaves and flowers was slower when they fed female than male leaves and flower buds. In addition, A. angulifera larvae fed fewer flower buds on female than on male plants. These results show that the male‐biased florivory on E. japonica trees is attributed mainly to the specialist florivore and also feeding preference for male flowers in an opportunistic florivore that feed both leaves and flowers.     

Effects of leaf and sap feeding insects on photosynthetic rates of goldenrod

Summary Herbivory can alter the balance between sources and sinks within a plant, and changes in the source-sink ratio often lead to changes in plant photosynthetic rates. We investigated how feeding by three insect herbivores affected photosynthetic rates and growth of goldenrod (Solidago altissima). One, a phloem-sap feeding aphid (Uroleucon caligatum), creates an additional sink, and the other two, a leaf-chewing beetle (Trirhabda sp.) and a xylem-sap feeding spittlebug (Philaenus spumarius) both reduce source supply by decreasing leaf area. Plants were grown outside in large pots and insects were placed on them at predetermined densities, with undamaged plants included as controls. All insects were removed after a 12-day feeding period. We measured photosynthetic rates both of damaged leaves and of undamaged leaves that were produced after insect removal. Photosynthetic rates per unit area of damaged leaves were reduced by spittlebug feeding, but not by beetle or aphid feeding. Conductance of spittlebugdamaged leaves did not differ from controls, but internal carbon dioxide concentrations were increased. These results indicate that spittlebug feeding does not cause stomatal closure, but impairs fixation within the leaf. Effects of spittlebug feeding on photosynthetic rates persisted after the insects were removed from the plants. Photosynthetic rates per unit area of leaves produced after insect removal on spittlegug-damaged plants were lower than control levels, even though the measurements were taken 12 days after insect removal. The measurement leaf on spittlebugdamaged plants was reduced in area by 27% relative to the controls, but specific leaf area (leaf area/leaf weight) was increased by 18%. Because of the shift in specific leaf area, photosynthetic rates were also examined per unit leaf weight, and when this was done there were no significant differences between control and spittlebug-damaged plants. Beetle and aphid feeding had no effects on the photosynthetic rate of the leaves produced after insect removal. Plant relative growth rates (in terms of height) were reduced by spittlebugs during the period that the insects were feeding on the plants. Relative growth rates of spittlebug-damaged plants were increased above control levels after insect removal, but these plants were still shorter than controls 17 days after insect removal. Beetles and aphids did not affect plant relative growth rates or plant height. Feeding by both spittlebugs and beetles reduced leaf area, and the effect of the spittlebug was more severe than that of the beetle. These results show that effects of herbivory on photosynthetic rates cannot be predicted simply by considering changes in the source-sink ratio, and that spittlebug feeding is more damaging to the host plant than beetle or aphid feeding.     

Genomic evidence of genetic variation with pleiotropic effects on caterpillar fitness and plant traits in a model legume

Plant–insect interactions are ubiquitous, and have been studied intensely because of their relevance to damage and pollination in agricultural plants, and to the ecology and evolution of biodiversity. Variation within species can affect the outcome of these interactions. Specific genes and chemicals that mediate these interactions have been identified, but genome‐ or metabolome‐scale studies might be necessary to better understand the ecological and evolutionary consequences of intraspecific variation for plant–insect interactions. Here, we present such a study. Specifically, we assess the consequences of genome‐wide genetic variation in the model plant Medicago truncatula for Lycaeides melissa caterpillar growth and survival (larval performance). Using a rearing experiment and a whole‐genome SNP data set (>5 million SNPs), we found that polygenic variation in M. truncatula explains 9%–41% of the observed variation in caterpillar growth and survival. Genetic correlations among caterpillar performance and other plant traits, including structural defences and some anonymous chemical features, suggest that multiple M. truncatula alleles have pleiotropic effects on plant traits and caterpillar performance (or that substantial linkage disequilibrium exists among distinct loci affecting subsets of these traits). A moderate proportion of the genetic effect of M. truncatula alleles on L. melissa performance can be explained by the effect of these alleles on the plant traits we measured, especially leaf toughness. Taken together, our results show that intraspecific genetic variation in M. truncatula has a substantial effect on the successful development of L. melissa caterpillars (i.e., on a plant–insect interaction), and further point toward traits potentially mediating this genetic effect.