Species-specific responses of herbivores to within-plant and environmentally mediated between-plant variability in plant chemistry

Allocation of resources to growth and defense against herbivores crucially affects plant competitiveness and survival, resulting in a specific distribution of assimilates and defense compounds within plant individuals. Additionally, plants rarely experience stable environmental conditions, and adaptations to abiotic and biotic stresses may involve shifts in resistance to herbivores. We studied the allocation of phytochemicals in Brassica oleracea (Brussels sprouts) due to leaf age, drought stress and herbivore damage and assessed effects on two lepidopteran herbivores differing in diet breadth: the generalist Spodoptera littoralis and the specialist Pieris brassicae. Glucosinolates as secondary defense compounds and total nitrogen and carbon were quantified and linked to plant palatability, i.e., herbivore feeding preference. Herbivore responses were highly species-specific and partially related to changes in phytochemicals. Spodoptera littoralis preferred middle-aged leaves with intermediate levels of glucosinolates and nitrogen over young, glucosinolate and nitrogen rich leaves, as well as over old leaves, poor in glucosinolates and nitrogen. In contrast, P. brassicae preferred young leaves. Both species preferred severely drought-stressed plants to the well-watered control, although analyzed glucosinolate concentrations did not differ. Both S. littoralis and P. brassicae feeding induced an increase of indole glucosinolate levels, which may explain a reduced consumption of damaged plants detected for S. littoralis but not for P. brassicae. By revealing distinct, sometimes contrasting responses of two insect herbivores to within-plant and stress-mediated intraspecific variation in phytochemistry of B. oleracea, this study emphasizes the need to consider specific herbivore responses to understand and predict the interactions between herbivores and variable plants.     

Dealing with double trouble: consequences of single and double herbivory in Brassica juncea

In their natural environment, plants are often attacked simultaneously by many insect species. The specificity of induced plant responses that is reported after single herbivore attacks may be compromised under double herbivory and this may influence later arriving herbivores. The present study focuses on the dynamics of induced plant responses induced by single and double herbivory, and their effects on successive herbivores. Morphological (leaf length, area and trichome density) and chemical changes (leaf alkenyl and indole glucosinolates) in Brassica juncea were evaluated 4, 10, 14 and 20 days after damage by the specialist Plutella xylostella alone, or together with the generalist Spodoptera litura. To assess the biological effect of the plant’s responses, the preference and performance of both herbivores on previously induced plants were measured. We found that alkenyl glucosinolates were induced 20 days after damage by P. xylostella alone, whereas their levels were elevated as early as 4 days after double herbivory. Trichome density was increased in both treatments, but was higher after double herbivory. Interestingly, there was an overall decrease in indole glucosinolates and an increase in leaf size due to damage by P. xylostella, which was not observed during double damage. S. litura preferred and performed better on undamaged plants, whereas P. xylostella preferred damaged plants and performed better on plants damaged 14 and 10 days after single and double herbivory, respectively. Our results suggest that temporal studies involving single versus multiple attacker situations are necessary to comprehend the role of induced plant responses in plant–herbivore interactions.     

The influence of metabolically engineered glucosinolates profiles in Arabidopsis thaliana on Plutella xylostella preference and performance

The oviposition preference and larval performance of the diamondback moth (DBM), Plutella xylostella, was studied using Arabidopsis thaliana plants with modified glucosinolate (GS) profiles containing novel GSs as a result of the introduction of individual CYP79 genes. The insect parameters were determined in a series of bioassays. The GS content of the plants as well as the number of trichomes were measured. Multivariate analysis was used to determine the possible relationships among insect and plant variables. The novel GSs in the tested lines did not appear to have any unequivocal effect on the DBM. Instead, the plant characteristics that affected larval performance and larval preference did not influence oviposition preference. Trichomes did not affect oviposition, but influenced larval parameters negatively. Although the tested A. thaliana lines had earlier been shown to influence disease resistance, in this study no clear results were found for P. xylostella.     

Effects of glucosinolate and myrosinase levels in Brassica juncea on a glucosinolate-sequestering herbivore – and vice versa

Summary. Larvae of the turnip sawfly, Athalia rosae L. (Hymenoptera: Tenthredinidae), sequester glucosinolates of their host plants, namely members of the Brassicaceae family, in their haemolymph. Therefore, they need to circumvent myrosinase activities of the plant tissue which normally hydrolyse the glucosinolates after plant damage. Effects of varying levels of glucosinolates and myrosinases on the performance of A. rosae were investigated using homozygous lines of Brassica juncea (L.) with either (1) low glucosinolate (lowGS) and low myrosinase (lowMR), (2) high glucosinolate (highGS) and high myrosinase (highMR), or (3) high glucosinolate (highGS) and low myrosinase (lowMR) levels. To insure that the given quantities remained as constant as possible, newly hatched larvae were enclosed on the second-youngest leaf of a plant, and were offered a new plant of comparable physiological age (6-leaf-stage) every day. The performance of A. rosae was little affected by leaf quality. Body masses of eonymphs and adults were on average lowest on the highGS/highMR-line, but these differences were rarely significant. The pupal developmental times of females and males were longest on the highGS/lowMR-line in only one of two replicate experiments. All other performance traits (developmental times of larvae, egg numbers, adult longevity) were not significantly different. Glucosinolates, sequestered by the larvae, are carried through the pupal stage. The glucosinolate concentration measured in adult insects reflected the level of the host plant line, without showing any obvious costs for sequestration. Obviously, A. rosae is highly tolerant to variation in the glucosinolate-myrosinase system of its host. In addition, induced changes of glucosinolate concentrations and myrosinase activities caused by 24 h-feeding of groups of three small larvae were analysed in the second-youngest leaves. In contrast to the patterns most herbivores evoke on Brassicaceae, namely an increase of both glucosinolate concentration and myrosinase activity, we detected a significant decrease of both traits in all three lines where the respective trait was originally high in the plants. Although glucosinolate levels dropped in the highGS lines about 50%, these still contained higher concentrations than the lowGS line. Whereas the activity of soluble myrosinases remained highest in the highMR line, even after a decrease to almost 30% due to feeding, the levels of insoluble myrosinases converged after feeding in lowMR and highMR lines. Levels of the signalling molecule salicylic acid slightly decreased on average after feeding, whereas jasmonic acid was below the detection threshold in almost all samples. The concentration of several molecules varies strongly in plant tissue with age and can change due to induction by herbivore feeding. Therefore, if performance of an insect species is measured on plants with specific traits, the variability in these traits needs to be carefully controlled in experiments.     

Impact of hydroxylated and non-hydroxylated aliphatic glucosinolates in Arabidopsis thaliana crosses on plant resistance against a generalist and a specialist herbivore

Glucosinolates (GSs) are part of a two-component defence system, characteristic for the Brassicales, including the model species Arabidopsis thaliana (L.) Heynh. The defence activity of GSs is associated with different side chain structures. The AOP genes are central in side-chain modification. AOP2 mediates formation of alkenyl GS from a methylsulfinyl precursor, whereas AOP3 catalyzes production of hydroxy-alkyl GSs from the same precursor. Although several studies have assessed the role of GSs in plant defence, the function of specific aliphatic GSs in plant defence is still not clarified. Structural different GSs may influence insect herbivores differentially. We created a set of plant lines derived of a cross between two A. thaliana accessions, Gie-0 × Sap-0, which dominantly accumulate either 3-methylsulfinylpropyl GS or 3-hydroxypropyl GS. The generalist Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) and the crucifer-specialist Pieris brassicae (L.) (Lepidoptera: Pieridae) were used as model insects, to study effects of individual aliphatic GSs on lepidopteran herbivores with a different feeding specialization. However, the experiments revealed that weight gain of S. exigua and P. brassicae third and fourth-larval instars was similar on both chemotypes. But leaf consumption of the generalist was higher on 3-methylsulfinylpropyl-producing lines with low GS levels (23.2 μmol g⁻¹) than on 3-hydroxypropyl-producing lines that contained a more than twofold higher amount of GSs (60 μmol g⁻¹). In contrast, no differential effects of non-hydroxylated and hydroxylated GSs were found on the specialist P. brassicae. Our study indicates that there is no simple relationship between GS content and insect responses.     

Local and systemic induced responses to cabbage root fly larvae (Delia radicum) in Brassica nigra and B. oleracea

Summary. Feeding by belowground herbivores may induce systemic changes in shoot defence levels that affect the performance of above ground herbivores and higher trophic levels. In this paper two wild Brassica species, B. nigra and B. oleracea were experimentally infested with 10 larvae of the cabbage root fly, Delia radicum. Plant dry masses and glucosinolate levels in shoots, main roots, and fine roots were determined at 3, 7, 12 and 14 days after infestation and compared to those of control plants. The systemic response in the leaves differed between plant species. In B. nigra shoot glucosinolate levels in D. radicum infested plants steadily increased with time until they were almost twice those of controls 14 days after infestation. B. oleracea plants infested with D. radicum did not show significant changes in shoot glucosinolate levels within 14 days, which may be due to the unexpected poorer performance of D. radicum on this species. Both plant species showed a local increase in indole glucosinolates in the main roots, which are the preferred feeding site of D. radicum larvae. B. oleracea plants however showed a stronger (1.9 – 4.7 times) increase in indole glucosinolate levels than B. nigra (1.5 – 2.6 times). The increase in indole glucosinolates in B. nigra main roots, was counterbalanced by a significant decrease in aromatic glucosinolate levels. These differences in local responses to D. radicum feeding between the two species may have contributed to the slower growth rates of the larvae on B. oleracea. D. radicum feeding did not result in altered glucosinolate levels in the fine roots in either plant species. The differences in glucosinolate induction patterns between the summer annual B. nigra and the perennial B. oleracea are discussed in the light of their different life histories.     

Chemosensory and behavioural responses of the turnip sawfly, Athalia rosae, to glucosinolates and isothiocyanates

Summary. The turnip sawfly Athalia rosae sequesters glucosinolates from its cruciferous host plants in the larval stage. Investigation of the chemosensory and behavioural responses of adult A. rosae to glucosinolates and their volatile hydrolysis products, isothiocyanates, revealed that females detect glucosinolates by contact chemoreception and isothiocyanates by antennal olfaction. In electroantennogram recordings, four isothiocyanates (allyl [2-propenyl] isothiocyanate, benzyl isothiocyanate, butyl isothiocyanate and iberverin [3-methylthiopropyl isothiocyanate]) were active at all doses presented, including the lowest (0.1 μg), whilst the threshold for detection of three others, iberin [3-methylsulphinylpropyl isothiocyanate], methyl isothiocyanate, and sulforaphane [4-methylsulphinylbutyl isothiocyanate], was higher, at between 1 and 10 μg (source concentration of volatiles). Allyl isothiocyanate attracted experienced females in a four-chambered olfactometer, whilst na?ve females showed no response. Allyl isothiocyanate also attracted mature females to baited yellow water traps in field trials, although immature females were repelled at high isothiocyanate concentrations. In laboratory behavioural bioassays the glucosinolates sinigrin (allyl [2-propenyl] glucosinolate) and sinalbin (p-hydroxybenzyl glucosinolate), stimulated ovipositor probing in mature female A. rosae to an extent comparable to hot-water extracts of their host plants. These responses show that glucosinolates and isothiocyanates play an important role in host finding and host recognition in A. rosae.     

Secondary metabolites of the leaf surface affected by sulphur fertilisation and perceived by the diamondback moth

Summary. Summary. Oilseed rape, Brassica napus L. (cv Express), plants were grown under three different sulphur regimes: sulphur-free (S₀), normal sulphur (S_n, normal field concentration) and a sulphur-rich (S₊, 2 × concentration of S_n). We performed dual choice oviposition assays with the diamondback moth, Plutella xylostella, using real plants and, for the first time with this insect, artificial leaves sprayed with methanolic leaf-surface extracts. The results mirrored those of a separate study of preferences for whole plants. Females laid more eggs on surrogate leaves that were treated with S_n extracts than on S₀ plants, while only a slight, not significant, difference was observed between extracts of normal and sulphur-rich plants. This shows that chemical compounds on the leaf surface mediate the oviposition preference and that the female insect can perceive the quality of the host-plants in terms of their fertilisation status.Since leaf volatiles are known to be oviposition stimulants, we investigated the effects of leaf-surface extracts on insect olfactory responses using electroantennograms (EAGs). In agreement with the behavioural data, we found that extracts of sulphur-treated plants yielded higher EAG amplitudes than the S₀ extracts. Since the leaf content of the volatiles isothiocyanates is influenced by sulphur nutrition, we analysed the extracts for these compounds. Above the detection threshold of our GC-MS system, no isothiocyanates were found. Thus, other compounds present in the surface extracts must be perceived by the antenna.However, the HPLC analysis revealed 11 different glucosinolates. Progoitrin (2-Hydroxy-3-butenyl) and gluconapoleiferin (2-Hydroxy-4-pentenyl), which belong to the hydroxy-alkene class of glucosinolates, were the most abundant compounds. The total glucosinolate content sharply increased from S₀ to S_n plants, whereas it was slightly lower in _n versus S₊ plants. Since it is known that glucosinolates can stimulate oviposition, it seems likely that the increased content we observed was influencing the insect preference in this study too.     

Responses of the flea beetles Phyllotreta nemorum and P. cruciferae to metabolically engineered Arabidopsis thaliana with an altered glucosinolate profile

Summary. Insects feeding on Cruciferae recognize their host plants at least partially by means of specific responses to glucosinolates. However, the effects of variations in glucosinolate levels on the acceptability of plants for specialized insects are not well understood. A survey of the literature demonstrated positive, no, as well as negative correlations between plant acceptability and glucosinolate levels. The present study took advantage of the presence of transgenic Arabidopsis thaliana plants with increased glucosinolate levels. Transgenic A. thaliana contain the CYP79A1 gene from Sorghum bicolor. This gene encodes an enzyme which converts L-tyrosine into p-hydroxyphenylacetaldoxime in the biosynthesis of cyanogenic glycosides in S. bicolor. In transgenic A. thaliana plants, endogenous enzymes convert p-hydroxyphenylacetaldoxime into p-hydroxybenzylglucosinolate (sinalbin), which is not found naturally in this plant. The introduction of CYP79A1 resulted in a four-fold increase in total glucosinolate levels in transgenic A. thaliana plants. Although these changes in glucosinolate levels were rather dramatic, they did not have any effects on the acceptability of A. thaliana for the two flea beetle species, Phyllotreta nemorum and P. cruciferae. The flea beetles did not discriminate between transgenic and wildtype plants. Furthermore, they did not discriminate between leaf discs of wildtype plants where different concentrations of p-hydroxybenzylglucosinolate had been applied topically on the leaf surface. Feeding in P. nemorum was stimulated by extremely high levels of allylglucosinolate while this compound had no effect on P. cruciferae. It is concluded that the effect of glucosinolates on adapted insects depends on the chemical or physical environment in which the glucosinolates are found.     

Multiple feeding stimulants in <Emphasis Type="Italic">Sinapis alba</Emphasis> for the oligophagous leaf beetle <Emphasis Type="Italic">Phaedon cochleariae</Emphasis>

Summary. In earlier investigations on host plant discrimination of leaf beetles glucosinolates were described as feeding stimulants for the Brassicaceae specialist Phaedon cochleariae F. (Coleoptera: Chrysomelidae). However, since these findings could not be confirmed in later studies offering 2-propenylglucosinolate in concentrations corresponding to those detected in host plant leaf material, the identification of feeding stimulants of this leaf beetle species remained unclear. In order to investigate which compounds of the host plant Sinapis alba (Brassicaceae) are involved in feeding stimulation, leaf extracts of different polarities were tested in bioassays with adults of P. cochleariae. Number of feeding beetles and net consumption rates were highest on pea leaves painted with methanol extracts of S. alba, whereas weak feeding responses were also detectable for hexane extracts. In subsequent bioassay-guided fractionations of methanol extracts with semi-preparative high performance liquid chromatography, two distinct fractions, one containing glucosinolates and another containing flavonoids, were found to stimulate beetles to feed to variable degrees. Other collected fractions had zero activity. The combination of both active fractions evoked significantly higher consumption rates and stimulated more beetles to feed than fractions tested individually. At least one compound of each fraction, among these the main glucosinolate of S. alba, 4-hydroxybenzylglucosinolate, act additively. Effects of two different naturally-occurring ratios of glucosinolates and flavonoids on the strength of feeding responses were investigated by use of extracts of two sets of host plants differently exposed to radiation. One set was outdoors-exposed, whereas the second set was kept in the greenhouse. However, the feeding behaviour of P. cochleariae was not affected by the significantly different relative compositions of both compound classes in the host material. In conclusion, mustard leaf beetles need a combination of distinct plant metabolites acting in concert for feeding stimulation, whereby the mere presence of these stimulants, but probably not the ratio of involved compounds, determines their feeding response.     

Lack of sequestration of host plant glucosinolates in <Emphasis Type="Italic">Pieris rapae</Emphasis> and <Emphasis Type="Italic">P. grarricae</Emphasis>

Summary. Sequestration of plant toxins in herbivores is often correlated with aposematic coloration and gregarious behaviour. Larvae of Pieris brassicae show these conspicuous morphological and behavioural characteristics and were thus suggested to sequester glucosinolates that are characteristic secondary metabolites of their host plants. P. rapaeare camouflaged and solitary, and are thus not expected to sequester. To test this hypothesis and to check the repeatabi-lity of a study that did report the presence of the glucosinolate sinigrin in P. brassicae, larvae were reared on three species of Brassicaceae (Sinapis alba, Brassica nigra and Barbarea stricta), and different leaf and insect samples were taken for glucosinolate analysis. The major host plant glucosinolates could only be found in traces or not at all in larval haemolymph, bled or starved larvae, faeces or pupae of both species or P. brassicae regurgitant. Haemolymph of both Pieris spp. was not rejected by the ant Myrmica rubra in dual-choice assays; the regurgitant of P. brassicae was rejected. This suggests the presence of compounds other than glucosinolates that might be sequestered in or produced by P. brassicae only. In faeces of both Pieris spp. a compound which yielded 4-hydroxybenzylcyanide (HBC) upon incubation with sulfatase was detected in high concentrations when larvae had been reared on S. alba. This compound may be derived from hydrolysis of sinalbin, the main glucosinolate of that plant. The unidentified HBC progenitor was apparently not sequestered in the two Pieris spp., and was not detected in faeces of larvae reared on B. nigra or B. stricta. Received 18 July 2002; accepted 11 September 2002.     

Resistance of the generalist moth Trichoplusia ni (Noctuidae) to a novel chemical defense in the invasive plant Conium maculatum

Summary. Conium maculatum is an apiaceous species native to Eurasia that is highly toxic to vertebrates due to the presence of piperidine alkaloids, including coniine and γ-coniceine. More than 200 years after invading the United States this species remains mostly free from generalist insect herbivores. The presence of novel chemical defenses in the introduced range could provide invasive species with a competitive advantage relative to native plants. The cabbage looper (Trichoplusia ni) is a generalist lepidopteran found throughout the US that occasionally feeds on C. maculatum. We evaluated the toxicity of piperidine alkaloids to T. ni and determined putative resistance mechanisms, both behavioral and physiological, that allows this insect to develop successfully on C. maculatum foliage. T. ni larvae raised on diets enriched with coniine and γ-coniceine showed a decrease in consumption and longer development time, but no effects on growth were found at any alkaloid concentration. In a diet choice experiment T. ni larvae showed no avoidance of alkaloid-enriched diets, suggesting that the deterrence produced by alkaloids was related to a post-ingestive metabolic response. The ability of T. ni to consume diets high in alkaloid content could be due to at least three different mechanisms: 1) a decreased consumption rate, 2) efficient excretion of at least 1/3 of ingested alkaloids unmetabolized in frass, and 3) partial detoxification of alkaloids by cytochrome P450 s, as shown by the decreased larval growth in the presence of piperonyl butoxide, a P450 inhibitor. Even though T. ni tolerates C. maculatum alkaloids, the use of this species as a host plant could be ecologically disadvantageous due to prolonged larval growth and thus increased exposure to predators. Novel plant secondary compounds do not guarantee increased resistance to generalist herbivores.     

Chemical characterization of contact semiochemicals for host-recognition and host-acceptance by the specialist parasitoid <Emphasis Type="Italic">Cotesia plutellae</Emphasis> (Kurdjumov)

Summary. Cotesia plutellae is a specialist parasitoid of Plutella xylostella. This specificity is potentially under the control of several factors before and after oviposition. Thereby, the stimuli that lead female parasitoids to host locations and to oviposition, might be at the basis of the specificity. We explore here the response of C. plutellae females exposed to host cuticular lipids. A total cuticular lipid extract of host caterpillars was fractionated into a hydrocarbon fraction and a non-hydrocarbon fraction. Neither fraction alone had any effect on oviposition behaviour in C. plutellae but the hydrocarbon fraction alone did seem to have a positive effect on the rate of antennal contact by the females. To induce oviposition behaviour, both fractions were necessary and reflect cooperation between at least one compound in each fraction. Identification of cuticular lipids shows that hydrocarbons were dominant (77%). Non-hydrocarbon compounds were mainly represented by 15-nonacosanone (18% of the total lipid extract). This ketone is rare in insect cuticle lipids and is thought to originate from the cabbage epicuticle where it is dominant with n-C₂₉ and 14- and 15-nonacosanol also found among the cuticular lipids of the host caterpillar.     

Integrating phenological, chemical and biotic defences in ant-plant protection mutualisms: a case study of two myrmecophyte lineages

Summary. We examined the role of plant phenology in the evolution of anti-herbivore defence in symbiotic ant-plant protection mutualisms. Phenology of the host-plant affects traits of its herbivores, including size, growth rate, development time, and gregariousness. Traits of herbivores in turn determine what traits ants must have to protect their host. Diversity in plant phenological traits could thus help explain the great ecological diversity of coevolved ant-plant mutualisms. We explored the postulated causal chain linking phenology of the plant, herbivore adaptations to phenology, and ant adaptations for protection, by comparing two myrmecophytes presenting strong contrasts in phenology. In Leonardoxa africana, a slow-growing understory tree, growth at each twig terminal is intermittent, the rapid flushing of a single leaf-bearing internode being followed by a pause of several months. In contrast, axes of Barteria nigritana, a tree of open areas, grow continuously. Analysis of the phenology (kinetics of expansion) and chemistry of leaf development (contents of chlorophylls, lignin, and nitrogen during leaf growth) showed that these two species exhibit strongly contrasting strategies. Leonardoxa exhibited a delayed greening strategy, with rapid expansion of leaves during a short period, followed by synthesis of chlorophylls and lignins only after final leaf size has been reached. In contrast, leaves of Barteria expanded more slowly, with chlorophylls and lignin gradually synthesised throughout development. Differences in the phenology of leaf development are reflected in differences in the duration of larval development, and thereby in size, of the principal lepidopteran herbivores observed on these two plants. This difference may in turn have led to different requirements for effective defence by ants. The strategy of phenological defence may thus affect the evolution of biotic defence.     

Pyrrolizidine alkaloids on three trophic levels – evidence for toxic and deterrent effects on phytophages and predators

Summary. Pyrrolizidine alkaloids (PAs) present a model system in the investigation of tritrophic interactions mediated by plant secondary compounds. However, their toxicity for insect herbivores has never been experimentally proven. Here, we demonstrate the toxic effects of a PA on growth and survival of the eri silk moth Philosamia ricini. In a feeding experiment, larvae of this generalist herbivore fed with an artificial PA diet gained weight significantly slower than control animals, and died as pupae. We suggest that derivatives of the ingested PA N-oxide damage developmental functions during metamorphosis. A tracer test with [¹⁴C]senecionine N-oxide revealed that the caterpillars lack adaptations that would prevent conversion of the chemical into the pro-toxic free base. In contrast, the PA adapted leaf beetle Longitarsus anchusae accumulates PAs as N-oxides. We tested the purpose of sequestration in this species as defence against predators. Through a series of prey choice experiments with three carabid predator species, chemically non-protected bark beetle pupae were chosen almost uniformly over L. anchusae pupae. In a following choice test with one of these predators, artificially PA-treated mealworm segments deterred the predator from feeding. Overall the study corroborates the immediate toxic effect of PAs on non-adapted herbivores and the protective effect that adapted insects may gain by sequestering them. It thereby underlines the potential for PAs to play a central role in multitrophic interactions between plants, phytophages and their predators.     

Direct and indirect effects of CO2, nitrogen, and community diversity on plant-enemy interactions

Resource abundance and plant diversity are two predominant factors hypothesized to influence the amount of damage plants receive from natural enemies. Many impacts of these environmental variables on plant damage are likely indirect and result because both resource availability and diversity can influence plant traits associated with attractiveness to herbivores or susceptibility to pathogens. We used a long-term, manipulative field experiment to investigate how carbon dioxide (CO2) enrichment, nitrogen (N) fertilization, and plant community diversity affect plant traits and the amount of herbivore and pathogen damage experienced by the common prairie legume Lespedeza capitata. We detected little evidence that CO2 or N affected plant traits; however, plants growing in high-diversity treatments (polycultures) were taller, were less pubescent, and produced thinner leaves (higher specific leaf area). Interestingly, we also detected little evidence that CO2 or N affect damage. Plants growing in polycultures compared to monocultures, however, experienced a fivefold increase in damage from generalist herbivores, 64% less damage from specialist herbivores, and 91% less damage from pathogens. Moreover, within diversity treatments, damage by generalist herbivores was negatively correlated with pubescence and often was positively correlated with plant height, while damage by specialist herbivores typically was positively correlated with pubescence and negatively associated with height. These patterns are consistent with changes in plant traits driving differences in herbivory between diversity treatments. In contrast, changes in measured plant traits did not explain the difference in disease incidence between monocultures and polycultures. In summary, our data provide little evidence that CO2 or N supply alter damage from natural enemies. By contrast, plants grown in monocultures experienced greater specialist herbivore and pathogen damage but less generalist herbivore damage than plants grown in diverse communities. Part of this diversity effect was mediated by changes in plant traits, many of which likely are plastic responses to diversity treatments, but some of which may be the result of evolutionary changes in response to these long-term experimental manipulations.     

Mother’s choice of the oviposition site: balancing risk of egg parasitism and need of food supply for the progeny with an infochemical shelter?

Summary. Oviposition site selection of herbivorous insects depends primarily on host plant presence which is essential for offspring survival. However, parasitoids can exploit host plant cues for host location. In this study, we hypothesised that herbivores can solve this dilemma by ovipositing within high plant diversity. A diverse plant species composition might represent an ‘infochemical shelter’, as a potentially complex volatile blend can negatively affect the host location ability of parasitoids. We examined this exemplarily for the egg-laying response of the generalist leaf beetle, Galeruca tanaceti, in relation to (1) host plant availability and (2) plant species diversity in the field. Further, we investigated the effect of odours from mixed plant species compositions on (3) leaf beetle oviposition site selection and on (4) the orientation of its specialised egg parasitoid, Oomyzus galerucivorus. In the field, egg clutch occurrence was positively related to the presence and quantity of two major host plants, Achillea millefolium (yarrow) and Centaurea jacea, and to the number of herbaceous plant species. In two-choice bioassays, female beetles oviposited more frequently on sites surrounded by an odour blend from a diverse plant species composition (including yarrow) than on sites with a pure grass odour blend. In the presence of yarrow odour and an odour blend from a diverse plant mixture (including yarrow) no difference in the oviposition response was recorded. Experienced parasitoid females were attracted to yarrow odours, but showed no response when yarrow odours were offered simultaneously with odours of a non-host plant. In conclusion, it could be shown in laboratory bioassays that the parasitoid responds only to pure host plant odours but not to complex odour blends. In contrast, the herbivore prefers to oviposit within diverse vegetation in the field and in the laboratory. However, the laboratory results also point to a priority of host plant availability over the selection of a potential ‘infochemical shelter’ for oviposition due to high plant diversity.     

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.     

Herbivores on a dominant understory shrub increase local plant diversity in rain forest communities

Indirect effects of trophic interactions on biodiversity can be large and common, even in complex communities. Previous experiments with dominant understory Piper shrubs in a Costa Rican rain forest revealed that increases in herbivore densities on these shrubs caused widespread seedling mortality as a result of herbivores moving from Piper to seedlings of many different plant genera. We tested components of the Janzen-Connell hypothesis by conducting focused studies on the effects of specialist and generalist Piper herbivores on local seedling diversity. Whereas specialist herbivores are predicted to increase mortality to neighboring seedlings that are closely related to the source plant, true generalists moving from source plants may cause density-dependent mortality of many species, and possibly increase richness if new species replace abundant species that have been thinned by herbivores. Therefore, we hypothesized that seedling richness would be greater in understory control plots created in patches of Piper that had normal densities of generalist herbivores compared to plots from which we removed generalist herbivores manually from all Piper shrubs. After 15 months, generalist-herbivore-removal plots had > 40% fewer seedlings, > 40% fewer species, and 40% greater seedling evenness, on average, than control plots with generalist herbivores intact. Using a complementary approach in unmanipulated plots in four forests, we used path analysis to test for a positive association between seedling diversity and herbivore damage on Piper species. In unmanipulated plots, for both generalist and specialist herbivores, our data were significant fits to the causal model that Piper herbivores decrease evenness and increase plant species richness, corroborating the experimental results. Because herbivores changed how individuals were apportioned among the species and families present (lower evenness), one interpretation of these associations between herbivores on Piper shrubs and local seedling richness is that high seedling mortality in dominant families allowed the colonization or survival of less common species. If interspecific or apparent competition allowed for a relative increase in species richness, then the Janzen-Connell hypothesis may extend its predictions to generalist seedling predators. We speculate that apparent competition may explain some of the deviations from neutral model predictions, especially at small scales.     

Variation in composition of predator-attracting allelochemicals emitted by herbivore-infested plants: Relative influence of plant and herbivore

Summary During foraging, natural enemies of herbivores may employ volatile allelochemicals that originate from an interaction of the herbivore and its host plant. The composition of allelochemical blends emitted by herbivore-infested plants is known to be affected by both the herbivore and the plant. Our chemical data add new evidence to the recent notion that the plants are more important than the herbivore in affecting the composition of the volatile blends. Blends emitted by apple leaves infested with spider mites of 2 different species,T. urticae andP. ulmi, differed less in composition (principally quantitative differences for some compounds) than blends emitted by leaves of two apple cultivars infested by the same spider-mite species,T. urticae (many quantitative and a few qualitative differences). Comparison between three plant species — apple, cucumber and Lima bean — reveals even larger differences between volatile blends emitted upon spider-mite damage (many quantitative differences and several qualitative differences).