Facultative sex ratio shifts by a herbivorous insect in response to variation in host plant quality

Summary We tested predictions of sex allocation theory with a series of field experiments on sex allocation in an herbivorous, haplodiploid, sawfly, Euura lasiolepis. Our experiments demonstrated the following points. 1) Adult females allocated progeny sex in response to plant growth. 2) Population sex ratios varied in response to plant quality, being male-biased where plant growth was slow and female-biased where plant growth was rapid. 3) Family sex ratios varied in response to plant quality, being male-biased on slow-growing plants and female-biased on rapidly-growing plants. 4) Female fitness increased more rapidly as the result of developing on more rapidly-growing plants than male mass. We conclude from these results that there are unequal returns on investment in male and female progeny. This results in facultatively biased sawfly sex ratios as an adaptive response to variation in plant quality.     

Effects of root herbivory by an insect on a foliar-feeding species,mediated through changes in the host plant

Summary The effects of root herbivory by larvae of the scarabaeid, Phyllopertha horticola, on the growth of Capsella bursa-pastoris were examined. Individuals of Aphis fabae were reared on the leaves to determine what effect, if any, root feeding has on the performance of this insect. The experiment was conducted under two watering regimes (low and high). Low watering and root feeding caused water stress in the plants and this was reflected in a reduction in vegetative biomass and an increase in the proportion of material allocated to reproduction. Supplying plants with ample water in the high treatment enabled the water stress caused by root herbivory to be offset, but not completely overcome. Low watering and root feeding caused an increase in aphid weight and growth rate, while root feeding also increased fecundity and adult longevity. These effects are attributed to an improvement in food quality, measured by total soluble nitrogen, and caused by amino acid mobilization due to the water stress. The implications of these results in agricultural and ecological situations are discussed.     

Copper and herbivory lead to priming and synergism in phytohormones and plant volatiles in the absence of salicylate-jasmonate antagonism

Abiotic stress factors can interfere with the emission of herbivore-induced plant volatile organic compounds (VOCs) and thus disrupt chemical communication channels between plants and other organisms. We investigated whether copper (Cu) stress alone or in conjunction with insect damage modifies the kinetics of (1) VOCs, (2) the VOC-inducing phytohormone jasmonic acid (JA) and (3) its putative antagonist salicylic acid (SA). Hydroponically grown Zea mays exposed to 10 and 80 µM of Cu showed no increases in JA or VOC levels in the absence of herbivory. However when challenged by herbivores, Cu (80 µM) caused ROS generation in root tissues and primed for increased JA accumulation and VOC emission in leaves. SA synthesis was equally primed but higher concentrations were also apparent before insects started feeding. In contrast, plants grown at 10 µM Cu did not differ from controls. These results show that abiotic and biotic stresses result in concentration-dependent, non-additive defense responses. Further support is given to the notion that JA-SA antagonism is absent in Z. mays.     

Inhibition of Telenomus sphingis an egg parasitoid of Manduca spp. by trichome/2-tridecanone-based host plant resistance in tomato

The resistance of accession PI 134417 of the wild tomato Lycopersicon hirsutum f. glabratum C. H. Mull to Manduca sexta (L.) (Lepidoptera: Spingidae) and Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae) is conditioned by the high densities of 2-tridecanone-containing, glandular trichomes associated with the foliage. In laboratory experiments, rates of parasitism of M. sexta eggs by Telenomus sphingis (Ashmead) (Hymenoptera: Scelionidae) were lower among eggs on PI 134417 foliage than among eggs on foliage of the cultivated tomato L. esculentum Mill. (cv. Better Boy). The latter is characterized by a significantly lower density of type VI glandular trichomes than PI 134417 and an absence of 2-tridecanone. Parasitism by T. sphingis was also reduced among eggs on foliage of the F₁ hybrid between PI 134417 and L. esculentum. The hybrid foliage lacks 2-tridecanone but has a density of type VI glandular trichomes that is intermediate between those of PI 134417 and L. esculentum, indicating that elevated densities of type VI glandular trichomes adversely affect T. sphingis. This conclusion was further substantiated by the finding that there were no differences among plant lines in the levels of parasitism of M. sexta eggs when the eggs were on foliage that had been divested of glandular trichomes.In bioassays in which T. sphingis adults or immatures in host eggs were exposed to filter paper treated with 2-tridecanone at rates comparable to those associated with PI 134417 foliage, 2-tridecanone was acutely toxic and caused high levels of mortality. In addition, at high concentrations, 2-tridecanone vapors were repellent to T. sphingis adults. However, when exposed to PI 134417 foliage, few T. sphingis adults were killed.Parasitism of M. sexta eggs was unaffected when the eggs were deposited by moths reared as fifth instar larvae on diet containing 2-tridecanone and/or 2-undecanone at levels comparable to those associated with PI 134417 foliage.     

Influence of an endophytic fungus on host plant selection by a polyphagous moth via volatile spectrum changes

Recently the role of micro-organisms as mediators of plant-herbivore interactions has been increasingly acknowledged in ecological research. We investigated the interaction between an unspecialized root fungal endophyte (Acremonium strictum) and the polyphagous moth Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), in greenhouse and laboratory bioassays. Specifically we examined in tomato (Lycopersicon esculentum Mill.) the systemic effects of the endophytic fungus on the host selection behaviour of female moths for oviposition and the volatile profiles of host plants to understand the mechanisms acting in this multi-trophic model system. Both laboratory and field strains of H. armigera moths oviposited more on leaves of A. strictum inoculated plants as compared to endophyte-free plants, both in free flight cages and in tethered moth laboratory experiments; the moth’s preferences were significant between 10 and 18 days after inoculation. The analysis of volatile profiles showed strong quantitative differences between treatments. Endophyte inoculated plants emitted diverse terpenes and sesquiterpenes at significantly lower amounts as compared to endophyte free-plants, except for α-terpinene, which did not differ between the treatments, and trans-β-caryophyllene, which was emitted in significantly higher amounts on inoculated plants. β-Thujene and α-phellandrene accounted for 73.3 and 12.0% of total amounts of volatiles emitted from endophyte-free and inoculated plants, respectively. Our findings demonstrate that A. strictum is able to systemically influence the host selection of H. armigera moths for oviposition; conceivably mediated by the induced changes in volatile emissions (and probably additional biochemical parameters of the host plants, which have not been analysed so far). We argue for a more detailed assessment of micro-organisms invisibly colonizing plants, when studying plant-herbivore or multitrophic interactions.     

Constraints on an induced defense: the role of leaf area

Folivory results in both leaf damage and the loss of photosynthetic capacity. Leaf damage activates the production of induced defenses, but diminished photosynthetic capacity resulting from lost leaf area may impair a plant's ability to respond defensively. Because damage-induced nicotine production in Nicotiana sylvestris (Solanaceae) is an energy-demanding, active process, we predicted that the loss of leaf area would constrain this plant's ability to produce an induced nicotine pool. We examined our prediction in an experiment which combined leaf puncture and removal protocols, quantified induced nicotine pools on a whole-plant basis, and accounted for losses in the nicotine pool due to removed leaves and lost growth potential. In contrast to our prediction, leaf removal did little to diminish the growth-corrected estimates of the induced nicotine pool in plants with sufficient damage cue; only when plants had lost 88% or more of their leaf area did the induced nicotine pool decline significantly. These results demonstrate that the induced defense is relatively insensitive to current photosynthetic capacity. In contrast to the size of the induced nicotine pool, the concentration of nicotine in the remaining shoot tissues continued to rise as puncture damage increased over all defoliation levels tested. The mechanisms responsible for inducible nicotine production may have evolved as a means of providing shoot tissues with protection that is proportional to the amount of damage incurred while keeping production costs constant for the remaining plant parts.     

Effects of plant competition and herbivore density on the development of the turnip root fly (Delia floralis) in an intercropping system

In this study, interactive effects of plant competition and herbivory on plant quality and herbivore development were examined in a greenhouse experiment where cabbage plants [Brassica oleracea L. var. capitata (Brassicaceae)] were intercropped with red clover [Trifolium pratense L. (Fabaceae)]. Cabbages were grown with two red clover densities and attack rates by the root feeding herbivore the turnip root fly, Delia floralis Fall. (Diptera: Anthomyiidae). Above ground and below ground cabbage biomass was reduced through intercropping and larval damage. Intercropping also resulted in lower nitrogen and higher carbon root levels compared with levels in the roots of monocultured cabbage. Furthermore, both root nitrogen and carbon levels increased with herbivory. Root neutral detergent fibre (NDF) and lignin content increased in response to both increased plant competition and higher egg densities. For lignin, an interaction effect was observed in the form of elevated levels in intercropped plants subjected to larval damage, while levels in roots of monocultured cabbage remained unchanged. The quality changes brought about by clover competition affected D. floralis development negatively, which resulted in reduced pupal weight. In addition, increased egg density also decreased larval growth. The effects on the development of D. floralis in relation to host plant quality are discussed. Handling editor: Gimme Walter     

Infection by the VA-mycorrhizal fungusGlomus caledonium inHedysarum coronarium as influenced by host plant and P content of soil

The low degree of infection ofHedysarum coronarium L. (sulla) exposed to inoculum of the VAM endophyteGlomus caledonium was investigated. Infection began after a prolonged lag phase and remained at very low levels even after three months’ growth. Neither very high rates of inoculum, nor very low P content of the soil raised the low infection level of the sulla plants. There appeared to be some differences in rate of infection among ten different ecotypes of sulla but the level of infection remained low in all cases. In all tested populations some plants remained uninfected. The low infection rate of sulla may therefore have a genetic basis. It was shown that the growth ofH. coronarium is hardly improved by phosphate fertilization. This may explain the poor response of this plant species, adapted to grow in nutrient-deficient soil, to VAM. Programmes aimed at increasing the productivity in marginal soils through the introduction of efficient VAM endophytes should take into account the fact that certain plant species growing in marginal soils may not always benefit from mycorrhizal inoculation, due to their inherently low mycorrhizal dependency.     

Fungi associated with the southern pine beetle: avoidance of induced defense response in loblolly pine

Summary Loblolly pines (Pinus taeda) are rapidly killed by colonizing southern pine beetle (Dendroctonus frontalis). The female beetles carry two species of fungi (Ceratocystis minor var. barrasii and an unnamed basidiomycete) within a mycangium. The insects are also frequently associated with a blue-staining form of C. minor. These fungi are inoculated into the tree during colonization. The tree has an induced defensive response that involves resin soaking and necrosis of affected tissue isolating the invading organlsms. The blue-staining fungus stimulates formation of this response in the tree, but the two mycangial fungi do not. These results suggest that the beetles are closely associated with two highly pathogenic fungi that do not stimulate one of the critical components of tree defense.     

Protection of apple against fire blight induced by an hrpL mutant of Erwinia amylovora

A regulatory hrpL non-virulent mutant of Erwinia amylovora is effective in controlling fire blight disease when inoculated on apple seedlings simultaneously with the pathogenic parental strain. Mechanisms involved in this protective effect were investigated. The use of two marker genes, uidA and lacZ, expressed in the hrpL mutant and the pathogenic strain, respectively, allowed to localize simultaneously the two inoculated strains in plant tissue. An anti-β-glucuronidase antibody was also used to detect the hrpL mutant. Both techniques indicated that the two strains localized mainly in separate areas of the leaf tissue. In addition, leaves infiltrated with the hrpL mutant exhibited a significant increase in peroxidase activity in contrast to a hrp secretion mutant known to be less effective in the protection. It is suggested that protection obtained with the hrpL mutant relies on the physical separation between the mutant and the parental strain after co-inoculation and the rapid and sustained activation of plant defense mechanisms in reactive tissue, i.e. not invaded by the virulent strain.     

The olfactory response of cabbage root fly Delia radicum to the host plant volatile allylisothiocyanate

The responses of gravid female cabbage root flies, Delia radicum (L.), to brassica odour and allylisothiocyanate (ANCS) presented in either a discrete plume or uniformly dispersed in air passing through a wind tunnel were investigated. Relatively fast, straight flights occurred in diffuse odour conditions, while in discrete plumes flights were slower with more frequent changes of direction.Flies maintained a constant ground speed and track angle over the straight legs of their flight tracks in two different wind speeds by changing air speed and course. As flies approached an odour source in a discrete plume ground speed was progressively decreased and frequency of turning increased but track angles remained constant. This was achieved by reducing air speed and course angle.Flies released at increasing distances from an odour source moved upwind towards it in a series of short flights, the number relating to distance.The results are discussed in the light of current theories of anemotactic odour source locations and a sequence of orientated behaviour for host plant finding is proposed.

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Les réactions olfactives de Delia radicum à l'allylisothiocyanate volatil émis par les plantes-hôtes

Résumé L'étude a porté sur les réactions de femelles fécondées de Delia radicum L. à l'odour de chou et à l'allylisothiocyanate (ANCS) offerts, soit sous forme d'un fin panache, soit uniformément répartis dans l'air traversant un tunnel à vent.Avec une odeur diffuse les vols étaient relativement rapides et rectilignes, tandis qu'avec un fin panache les vols étaient plus lents avec des changements de direction plus fréquents.En présence de deux vitesses du courant d'air les mouches ont maintenu leur vitesse au sol et leur angle de vol par rapport à la direction du déplacement constants en modifiant leur vitesse de vol et leur direction.Quand les mouches approchaient de la source odorante dans un fin panache, la vitesse au sol diminuait progressivement et la fréquence des virages augmentait, les angles de vol restant constant. Ceci était obtenu en réduisant de la vitesse de l'air et l'angle du déplacement.Les mouches paraissent utiliser une anémotaxie optomotrice et mécanique pendant la localisation de l'hôte. Les résultats sont discutés à la lumière des théories classiques sur la localisation anémotactique des sources odorantes.

     

Performance of nymphal and adult southern green stink bug on an overwintering host plant and impact of nymph to adult food-switch

Performance of nymphs and adults of the southern green stink bug,Nezara viridula (L.) (Heteroptera: Pentatomidae) feeding on immature fruits of radish,Raphanus raphanistrum L. (Brassicaceae) was poor compared with feeding on immature soybean [Glycine max (L.) Merrill] pods, but a change from radish to soybean from nymph to adult alleviated some of the detrimental effects of this food on adult performance. Over 50% of nymphs died on radish, while only 15% died on soybean. To complete nymph development (stadia 2 through 5), male and female nymphs required 9.5 and 13.3 days longer on radish compared to soybean. Fresh body weight of 1-d-old adult females and males was ≅65% greater for those reared as nymphs on soybean than on radish, with females being significantly heavier than males on both foods. Survivorship and longevity of females, however, was greater for those fed on radish as nymphs and as adults, as compared to those that fed on soybean or radish followed by soybean. Males lived longer when fed exclusively on soybean. In general, reproductive performance and percentage gain in adult body weight was greatest for insects fed on soybean only, and were better for those whose food was switched from nymph to adult, as compared to those insects fed on radish solely.     

Baifuzi reduces transient ischemic brain damage through an interaction with the STREX domain of BKCa channels

Stroke is a long-term disability and one of the leading causes of death. However, no successful therapeutic intervention is available for the majority of stroke patients. In this study, we explored a traditional Chinese medicine Baifuzi (Typhonium giganteum Engl.). We show, at first, that the ethanol extract of Baifuzi exerts neuroprotective effects against brain damage induced by transient global or focal cerebral ischemia in rats and mice. Second, the extract activated large-conductance Ca²⁺-activated K⁺ channel (BK_Ca) channels, and BK_Ca channel blockade suppressed the neuroprotection of the extract, suggesting that the BK_Ca is the molecular target of Baifuzi. Third, Baifuzi cerebroside (Baifuzi-CB), purified from its ethanol extract, activated BK_Ca channels in a manner similar to that of the extract. Fourth, the stress axis hormone-regulated exon (STREX) domain of the BK_Ca channel directly interacted with Baifuzi-CB, and its deletion suppressed channel activation by Baifuzi-CB. These results indicate that Baifuzi-CB activated the BK_Ca channel through its direct interaction with the STREX domain of the channel and suggests that Baifuzi-CB merits exploration as a potential therapeutic agent for treating brain ischemia.     

Identification of PprM: a modulator of the PprI-dependent DNA damage response in Deinococcus radiodurans

Deinococcus radiodurans possesses a DNA damage response mechanism that acts via the PprI protein to induce RecA and PprA proteins, both of which are necessary in conferring extreme radioresistance. In an effort to further delineate the nature of the DNA damage response mechanism in D. radiodurans, we set out to identify novel components of the PprI-dependent signal transduction pathway in response to radiation stress. Here we demonstrate the discovery of a novel regulatory protein, PprM (a modulator of the PprI-dependent DNA damage response), which is a homolog of cold shock protein (Csp). Disruption of the pprM gene rendered D. radiodurans significantly sensitive to γ-rays. PprM regulates the induction of PprA but not that of RecA. PprM belongs in a distinct clade of a subfamily together with Csp homologs from D. geothermalis and Thermus thermophilus. Purified PprM is present as a homodimer under physiological conditions, as the case with Escherichia coli CspD. The pprA pprM double-disruptant strain exhibited higher sensitivity than the pprA or pprM single disruptant strains, suggesting that PprM regulates other hitherto unknown protein(s) important for radioresistance besides PprA. This study strongly suggests that PprM is involved in the radiation response mediated by PprI in D. radiodurans.     

Light-induced changes in the photoresponses of plant stems the loss of a high irradiance response to far-red light

De-etiolation results in phytochrome destruction, greening, and the loss of the far-red high irradiance responses (HIR). Evidence is presented against the hypothesis that the loss of the far-red HIR is a direct consequence of phytochrome destruction. Loss of the far-red HIR for the inhibition of elongation in hypocotyls of Raphanus sativus involves two different, but linked, actions of phytochrome. An induction reaction requires the far-red absorbing form of phytochrome for about 20 min after which accumulation of its product depends only on time. A second reaction requires continuous light or frequent short irradiations and involves cycling of the phytochrome system. This acts on the product of the induction reaction. It is proposed that in green plants an important mode of operation of phytochrome in the light depends on pigment cycling, and that during de-etiolation this system is established under phytochrome control.Abbreviations HIR high irradiance response - R red - FR farred light - P_tot phytochrome, P_r its red absorbing form, P_fr its far-red absorbing form A.M. Jose was the holder of Ministry of Agriculture, Fisheries and Food award AE 6819     

Role of plant volatiles in host location by the specialist parasitoidMicroplitis croceipes cresson (Braconidae: Hymenoptera)

A study was conducted to determine the primary source of volatile cues within the plant-host complex used by hostseeking freeflying female Microplitis cro-ceipesCresson in flight tunnel bioassays. In single-source and two-choice tests, using wasps given an oviposition experience on either cotton (Gossypium hirsutum)or cowpea (Vigna unguiculata)seedlings damaged by corn earworm (CEW; Helicoverpa zeaBoddie), the damaged seedlings were significantly more attractive than the CEW frass, which was in turn more attractive than the larvae themselves. In a series of two-choice wind-tunnel tests, the discriminatory ability of the wasps was examined, following various oviposition experiences. Significantly more wasps flew to plants with old damage than to plants with fresh damage, regardless of whether they had experience on fresh or old damage. In a comparison of plant species, wasps with only one experience on either hostdamaged cotton or host-damaged cowpea were unable to distinguish between them, and showed no preference for either plant, whereas wasps with multiple experiences on a particular plant preferentially flew to that plant in the choice test. In comparing hosts with nonhosts, wasps successfully learned to distinguish CEW from beet armyworm (BAW; Spodoptera exigua)on cotton but were unable to distinguish CEW from either BAW or cabbage looper (Trichoplusia ni)on cowpea. The results show the important role played by plant volatiles in the location of hosts by M. croceipesand indicate the wasps limitations in discriminating among the various odors. The ecological advantages and disadvantages of this behavior are discussed.     

Effects of simulated herbivory and resource availability on the invasive plant, Alternanthera philoxeroides in different habitats

In biological control programs, the insect natural enemy’s ability to suppress the plant invader may be affected by abiotic factors, such as resource availability, that can influence plant growth and reproduction. Understanding plant tolerance to herbivory under different environmental conditions will help to improve biocontrol efficacy. The invasive alligator weed (Alternanthera philoxeroides) has been successfully controlled by natural enemies in many aquatic habitats but not in terrestrial environments worldwide. This study examined the effects of different levels of simulated leaf herbivory on the growth of alligator weed at two levels of fertilization and three levels of soil moisture (aquatic, semi-aquatic, and terrestrial habitats). Increasing levels of simulated (manual) defoliation generally caused decreases in total biomass in all habitats. However, the plant appeared to respond differently to high levels of herbivory in the three habitats. Terrestrial plants showed the highest below–above ground mass ratio (R/S), indicating the plant is more tolerant to herbivory in terrestrial habitats than in aquatic habitats. The unfertilized treatment exhibited greater tolerance than the fertilized treatment in the terrestrial habitat at the first stage of this experiment (day 15), but fertilizer appears not to have influenced tolerance at the middle and last stages of the experiment. No such difference was found in semi-aquatic and aquatic habitats. These findings suggest that plant tolerance is affected by habitats and soil nutrients and this relationship could influence the biological control outcome. Plant compensatory response to herbivory under different environmental conditions should, therefore, be carefully considered when planning to use biological control in management programs against invasive plants.     

Evidence of DNA replication in an arbuscular mycorrhizal fungus in the absence of the host plant

Summary This study provides evidence thatGigaspora margarita replicates its nuclear DNA, even in the absence of a host plant. Three experimental approaches were used: (i) static cytofluorimetry to quantify the DNA content, (ii) pulse treatments with bromodeoxyuridine (BrdU), which is an analogue of thymidine, to reveal nuclei undergoing DNA synthesis, and (iii) ultrastructural observations to study changes in chromatin morphology during the fungal cell cycle. A slight second peak of approximately twice the value of a major peak was found by cytofluorimetry, showing that a small number of nuclei had entered in cycle during in vitro development. Nuclei which had incorporated BrdU were observed after pulses of 24 h; nuclei with condensed chromatin were also apparent at this time. The results demonstrate thatG. margarita has all the metabolic pathways needed to replicate its nuclear DNA even in the absence of the host, suggesting that more complex mechanisms inhibit the extended growth in vitro of arbuscular mycorrhizal fungi.Abbreviations AM-fungi arbuscular mycorrhizal fungi - A.U. arbitrary units - BrdU 5-bromo-2-deoxyuridine - DAPI 4,6-diamidino-2-phenylindole - UV ultraviolet light     

Evaluating the post-release efficacy of invasive plant biocontrol by insects: a comprehensive approach

We propose a comprehensive program to evaluate the post-release phase of biocontrol programs that use insect herbivores to control invasive plant species. We argue that any release should be done in randomized release and non-release sites and should be followed up by well-replicated sampling and experimental protocols that evaluate the degree of success or failure. These follow-up studies should include landscape scale monitoring across relevant habitat gradients of (1) the abundance of the biocontrol agent, (2) the impact of the biocontrol agent on the target plant species, (3) the potential for non-target effects, and (4) the response of native species and communities to a reduction in the invasive species. We also argue that (5) experimental reductions of the biocontrol agent are required to eliminate the chance that the putative impact of the biocontrol agent is not confounded with other causes. Finally, we describe six scenarios, informed largely by a community ecology perspective, in which a biocontrol agent may decrease the abundance or vigor of the target plant species but not lead to successful control where native communities re-establish. We classify these failure scenarios as either direct or indirect effects of the invasive plant species: Native Source Limitation, Static Competitive Hierarchies, Novel Weapons, Trophic Shifts, Invasive Engineering and Associated Invasives. Overall, we argue that well replicated and landscape-scale post release monitoring programs are required not only to evaluate critically the degree of success and failure of biocontrol programs worldwide but also to provide insights into improving future biocontrol efforts. Handling Editor: Heikki Hokkanen.     

Nickel hyperaccumulation as an anti-herbivore trait: considering the role of tolerance to damage

Metal hyperaccumulation is a striking trait exhibited by many plant species, but the evolutionary ecology of metal hyperaccumulation is poorly understood. It has been widely hypothesized that metal hyperaccumulation evolved to protect plants from herbivory. However, there is currently little evidence that metal hyperaccumulation enhances the fitness of plants in the presence of herbivory. In this study, we conducted a multi-factor greenhouse experiment to examine the effects of two soil nickel concentrations (unamended (0 μg/g) and nickel amended (600 μg/g)), and three levels of artificial damage (0, 10 and 50%) on the growth of plants from two populations of Thlaspi montanum var. montanum. We observed a significant interaction between soil nickel and artificial damage. An a posteriori analysis of this interaction revealed that the presence of nickel significantly improved the ability of T. montanum to tolerate the negative effects of intense damage. Our results indicate that metal hyperaccumulation could benefit T. montanum by increasing its tolerance to damage. This study suggests that there is a potential for the evolution of metal hyperaccumulation in response to intense herbivory on T. montanum.