Search

In natural and agricultural ecosystems, plants are often simultaneously or sequentially exposed to combinations of stressors. Here we tested whether limited water availability (LWA) affects plant response to insect herbivory using two populations of Eruca sativa from desert and Mediterranean habitats that differ in their induced defenses. Considering that such differences evolved as responses to biotic and possibly abiotic stress factors, the two populations offered an opportunity to study ecological aspects in plant response to combined stresses. Analysis of chemical defense mechanisms showed that LWA significantly induced total glucosinolate concentrations in the Mediterranean plants, but their concentrations were reduced in the desert plants. However, LWA, with and without subsequent jasmonate elicitation, significantly induced the expression of proteinase inhibitor in the desert plants. Results of a no-choice feeding experiment showed that LWA significantly increased desert plant resistance to Spodoptera littoralis larvae, whereas it did not affect the relatively strong basal resistance of the Mediterranean plants. LWA and subsequent jasmonate elicitation increased resistance against the generalist insect in Mediterranean plants, possibly due to both increased proteinase inhibitor expression and glucosinolate accumulation. The effect of LWA on the expression of genes involved in phytohormone signaling, abscisic acid (ABA-1) and jasmonic acid (AOC1), and the jasmonate responsive PDF1.2, suggested the involvement of abscisic acid in the regulation of defense mechanisms in the two populations. Our results indicate that specific genotypic responses should be considered when estimating general patterns in plant response to herbivory under water deficiency conditions.

Erysimum cheiranthoides L (Brassicaceae; wormseed wallflower) accumulates not only glucosinolates, which are characteristic of the Brassicaceae, but also abundant and diverse cardenolides. These steroid toxins, primarily glycosylated forms of digitoxigenin, cannogenol, and strophanthidin, inhibit the function of essential Na+/K+-ATPases in animal cells. We screened a population of 659 ethylmethanesulfonate-mutagenized E. cheiranthoides plants to identify isolates with altered cardenolide profiles. One mutant line exhibited 66% lower cardenolide content, resulting from greatly decreased cannogenol and strophanthidin glycosides, partially compensated for by increases in digitoxigenin glycosides. This phenotype was likely caused by a single-locus recessive mutation, as evidenced by a wildtype phenotype of F1 plants from a backcross, a 3:1 wildtype:mutant segregation in the F2 generation, and genetic mapping of the altered cardenolide phenotype to one position in the genome. The mutation created a more even cardenolide distribution, decreased the average cardenolide polarity, but did not impact most glucosinolates. Growth of generalist herbivores from two feeding guilds, Myzus persicae Sulzer (Hemiptera: Aphididae; green peach aphid) and Trichoplusia ni Hubner (Lepidoptera: Noctuidae; cabbage looper), was decreased on the mutant line compared to wildtype. Both herbivores accumulated cardenolides in proportion to the plant content, with T. ni accumulating higher total concentrations than M. persicae. Helveticoside, a relatively abundant cardenolide in E. cheiranthoides, was not detected in M. persicae feeding on these plants. Our results support the hypothesis that increased digitoxigenin glycosides provide improved protection against M. persicae and T. ni, despite an overall decrease in cardenolide content of the mutant line.

Plants defend themselves against herbivore attack by constitutively producing toxic secondary metabolites, as well as by inducing them in response to herbivore feeding. Induction of secondary metabolites can cross plant tissue boundaries, such as from root to shoot. However, whether the potential for plants to systemically induce secondary metabolites from roots to shoots shows genetic variability, and thus, potentially, is under selection conferring fitness benefits to the plants is an open question. To address this question, we induced 26 maternal plant families of the wild species Cardamine hirsuta belowground (BG) using the wound-mimicking phytohormone jasmonic acid (JA). We measured resistance against a generalist (Spodoptera littoralis) and a specialist (Pieris brassicae) herbivore species, as well as the production of glucosinolates (GSLs) in plants. We showed that BG induction increased AG resistance against the generalist but not against the specialist, and found substantial plant family-level variation for resistance and GSL induction. We further found that the systemic induction of several GSLs tempered the negative effects of herbivory on total seed set production. Using a widespread natural system, we thus confirm that BG to AG induction has a strong genetic component, and can be under positive selection by increasing plant fitness. We suggest that natural variation in systemic induction is in part dictated by allocation trade-offs between constitutive and inducible GSL production, as well as natural variation in AG and BG herbivore attack in nature.

The effects of temporal variation in the quality of short-lived annual plants on oviposition preference and larval performance of insect herbivores has thus far received little attention. This study examines the effects of plant age on female oviposition preference and offspring performance in the large cabbage white butterfly Pieris brassicae. Adult female butterflies lay variable clusters of eggs on the underside of short-lived annual species in the family Brassicaceae, including the short-lived annuals Brassica nigra and Sinapis arvensis, which are important food plants for P. brassicae in The Netherlands. Here, we compared oviposition preference and larval performance of P. brassicae on three age classes (young, mature, and pre-senescing) of B. nigra and S. arvensis plants. Oviposition preference of P. brassicae declined with plant age in both plant species. Whereas larvae performed similarly on all three age classes in B. nigra, preference and performance were weakly correlated in S. arvensis. Analysis of primary (sugars and amino acids) and secondary (glucosinolates) chemistry in the plant shoots revealed that differences in their quality and quantity were more pronounced with respect to tissue type (leaves vs. flowers) than among different developmental stages of both plant species. Butterflies of P. brassicae may prefer younger and smaller plants for oviposition anticipating that future plant growth and size is optimally synchronized with the final larval instar, which contributes >80% of larval growth before pupation. Electronic supplementary material The online version of this article (doi:10.1007/s10886-017-0853-9) contains supplementary material, which is available to authorized users.

We investigated the influences of two structurally similar glucosinolates, phenethylglucosinolate (gluconasturtiin, NAS) and its (S)-2-hydroxyl derivative glucobarbarin (BAR), as well as their hydrolysis products on larvae of the generalist Mamestra brassicae (Lepidoptera: Noctuidae). Previous results suggested a higher defensive activity of BAR than NAS based on resistance toward M. brassicae larvae of natural plant genotypes of Barbarea vulgaris R. Br. (Brassicaceae) dominated by BAR. In the present study, the hypothesis of a higher defensive activity of BAR than NAS was tested by comparing two Barbarea species similarly dominated either by BAR or by NAS and by testing effects of isolated BAR and NAS on larval survival and feeding preferences. Larvae reared on leaf disks of B. verna (Mill.) Asch. had a lower survival than those reared on B. vulgaris P- and G-chemotypes. Leaves of B. verna were dominated by NAS, whereas B. vulgaris chemotypes were dominated by BAR or its epimer. In addition, B. verna leaves showed a threefold higher activity of the glucosinolate-activating myrosinase enzymes. The main product of NAS from breakdown by endogenous enzymes including myrosinases ("autolysis") in B. verna leaves was phenethyl isothiocyanate, while the main products of BAR in autolyzed B. vulgaris leaves were a cyclized isothiocyanate product, namely an oxazolidine-2-thione, and a downstream metabolite, an oxazolidin-2-one. The glucosinolates BAR and NAS were isolated and offered to larvae on disks of cabbage. Both glucosinolates exerted similar negative effects on larval survival but effects of NAS tended to be more detrimental. Low concentrations of BAR, but not of NAS, stimulated larval feeding, whereas high BAR concentrations acted deterrent. NAS only tended to be deterrent at the highest concentration, but the difference was not significant. Recoveries of NAS and BAR on cabbage leaf disks were similar, and when hydrolyzed by mechanical leaf damage, the same isothiocyanate-type products as in Barbarea plants were formed with further conversion of BAR to cyclic products, (R)-5-phenyloxazolidine-2-thione [(R)-barbarin] and (R)-5-phenyloxazolidin-2-one [(R)-resedine]. We conclude that a previously proposed generally higher defensive activity of BAR than NAS to M. brassicae larvae could not be confirmed. Indeed, the higher resistance of NAS-containing B. verna plants may be due to a combined effect of rather high concentrations of NAS and a relatively high myrosinase activity or other plant traits not investigated yet.

In some plant-insect interactions, specialist herbivores exploit the chemical defenses of their food plant to their own advantage. Brassica plants produce glucosinolates that are broken down into defensive toxins when tissue is damaged, but the specialist aphid, Brevicoryne brassicae, uses these chemicals against its own natural enemies by becoming a "walking mustard-oil bomb". Analysis of glucosinolate concentrations in plant tissue and associated aphid colonies reveals that not only do aphids sequester glucosinolates, but they do so selectively. Aphids specifically accumulate sinigrin to high concentrations while preferentially excreting a structurally similar glucosinolate, progoitrin. Surveys of aphid infestation in wild populations of Brassica oleracea show that this pattern of sequestration and excretion maps onto host plant use. The probability of aphid infestation decreases with increasing concentrations of progoitrin in plants. Brassica brassicae, therefore, appear to select among food plants according to plant secondary metabolite profiles, and selectively store only some compounds that are used against their own enemies. The results demonstrate chemical and behavioral mechanisms that help to explain evidence of geographic patterns and evolutionary dynamics in Brassica-aphid interactions.

Garlic mustard is a well-known invader of deciduous forests of North America, yet the influence of environmental factors on garlic mustard allelochemical production is not well understood. Three experiments were conducted to detect interactions between one garlic mustard allelochemical (glucosinolate) production and light availability. First, to detect patterns of glucosinolate production across a natural light gradient, leaves and roots of mature plants and first-year rosettes were sampled in patches ranging from 100 to 2 % of full sun within an Indiana forest. Second, to determine whether genetic variation drives observed correlations between glucosinolate content and light, seed collected across light gradients within six sites was grown in a common garden and glucosinolate production was measured. Finally, to understand whether local adaptation occurred in garlic mustard's response to light, seed collected from defined light environments across six sites was grown under four light treatments. Results of the field sampling showed that mature plants' root glucosinolate content was elevated in high compared to low light. In the common garden experiment, however, there was no correlation between light availability at seed origin and constitutive glucosinolate content. Additionally, in the common light treatments, there was no evidence for local adaptation to light environment. Overall, the results indicate that plasticity in response to light, not genetic variation among plants growing in different light environments, generates correlations between glucosinolate content and light in the field. Since mature garlic mustard populations in high light may exhibit increased glucosinolate content, it makes them potential targets for management.

The performance and behavior of herbivores is strongly affected by the quality of their host plants, which is determined by various environmental conditions. We investigated the performance and preference of the polyphagous shoot-infesting aphid Myzus persicae on the host-plant Arabidopsis thaliana in a two-factorial design in which nitrate fertilization was varied by 33 %, and the root-infesting cyst-nematode Heterodera schachtii was present or absent. Aphid performance was influenced by these abiotic and biotic factors in an interactive way. Nematode presence decreased aphid performance when nitrate levels were low, whereas nematode infestation did not influence aphid performance under higher nitrate fertilization. Aphids followed the "mother knows best" principle when given a choice, settling preferentially on those plants on which they performed best. Hence, they preferred nematode-free over nematode-infested plants in the low fertilization treatment but host choice was not affected by nematodes under higher nitrate fertilization. The amino acid composition of the phloem exudates was significantly influenced by fertilization but also by the interaction of the two treatments. Various glucosinolates in the leaves, which provide an estimate of phloem glucosinolates, were not affected by the individual treatments but by the combination of fertilization and herbivory. These changes in primary and secondary metabolites may be decisive for the herbivore responses. Our data demonstrate that abiotic and biotic factors can interactively affect herbivores, adding a layer of complexity to plant-mediated herbivore interactions.

Due to global warming, species are expanding their range to higher latitudes. Some range expanding plants have become invasive in their new range. The Evolution of Increased Competitive Ability (EICA) hypothesis and the Shifting Defense Hypothesis (SDH) predict altered selection on plant defenses in the introduced range of invasive plants due to changes in herbivore pressures and communities. Here, we investigated chemical defenses (glucosinolates) of five native and seven invasive populations of the Eurasian invasive range expanding plant, Rorippa austriaca. Further, we studied feeding preferences of a generalist and a specialist herbivore among the populations. We detected eight glucosinolates in the leaves of R. austriaca. 8-Methylsulfinyloctyl glucosinolate was the most abundant glucosinolate in all plants. There were no overall differences between native and invasive plants in concentrations of glucosinolates. However, concentrations among populations within each range differed significantly. Feeding preference between the populations by a generalist herbivore was negatively correlated with glucosinolate concentrations. Feeding by a specialist did not differ between the populations and was not correlated with glucosinolates. Possibly, local differences in herbivore communities within each range may explain the differences in concentrations of glucosinolates among populations. Little support for the predictions of the EICA hypothesis or the SDH was found for the glucosinolate defenses of the studied native and invasive R. austriaca populations. © 2014 Springer Science+Business Media New York.

Plants of the Brassicaceae are defended from feeding by generalist insects by constitutively-expressed and herbivory-induced glucosinolates (GS). We induced Arabidopsis plants 1, 16 and 24 h prior to allowing neonate larvae of the generalist Helicoverpa armigera to feed on whole plants for 72 h. These plants were subsequently retested with another group of neonates for a further 72 h. We used wild-type A. thaliana Col-0, and mutant lines lacking indolic GS, aliphatic GS or all GS. We hypothesized that larvae would not grow well on defended plants (WT) compared to those lacking GS, and would not grow well if plants had been primed or fed on for longer, due to the expected induced GS. There was survivorship on all lines suggesting H. armigera is a suitable generalist for these experiments. Larvae performed less well on wild-type and no indolic lines than on no aliphatic and no GS lines. Larvae distributed feeding damage extensively in all lines, more so on wild type and no-indolic lines. Contrary to expectations, larvae grew better on plants that had been induced for 1 to 16 h than on un-induced plants suggesting they moved to and selected less toxic plant parts within a heterogeneously defended plant. Performance declined on all lines if plants had been induced for 24 h, or had been fed upon for a further 72 h. However, contrary to expectation, individual and total GS did not increase after these two treatments. This suggests that Arabidopsis plants induce additional (not GS) defenses after longer induction periods.

Generalist insects show reduced selectivity when subjected to similar, but not identical, host plant chemical signatures. Here, we produced transgenic Arabidopsis thaliana plants that over-express genes regulating the aliphatic- and indolyl- glucosinolates biosynthetic pathways with either a constitutive (CaMV 35S) or a phloem-specific promoter (AtSUC2). This allowed us to examine how exposure to high levels of aliphatic- or indolyl-glucosinolates in homogenous habitats (leaf cage apparatus containing two wild-type or two transgenic leaves) and heterogeneous habitats (leaf cage apparatus containing one wild-type and one transgenic leaf) affects host selection and performance of Bemsia tabaci, a generalist phloem-feeding insect. Data from homogenous habitats indicated that exposure to A. thaliana plants accumulating high levels of aliphatic- or indolyl-glucosinolates negatively affected the performance of both adult females and nymphs of B. tabaci. Data from heterogeneous habitats indicated that B. tabaci adult females selected for oviposition plants on which their offspring perform better (preference-performance relationship). However, the combinations of wild-type and transgenic plants in heterogeneous habitats increased the period of time until the first choice was made and led to increased movement rate on transgenic plants, and reduced fecundity on wild-type plants. Overall, our findings are consistent with the view that both performance and selectivity of B. tabaci decrease in heterogeneous habitats that contain plants with closely-related chemical signatures.

Aboveground and belowground herbivore species modify plant defense responses differently. Simultaneous attack can lead to non-additive effects on primary and secondary metabolite composition in roots and shoots. We previously found that aphid (Brevicoryne brassicae) population growth on Brassica oleracea was reduced on plants that were infested with nematodes (Heterodera schachtii) prior (4 weeks) to aphid infestation. Here, we examined how infection with root-feeding nematodes affected primary and secondary metabolites in the host plant and whether this could explain the increase in aphid doubling time from 3.8 to 6.7 days. We hypothesized that the effects of herbivores on plant metabolites would depend on the presence of the other herbivore and that nematode-induced changes in primary metabolites would correlate with reduced aphid performance. Total glucosinolate concentration in the leaves was not affected by nematode presence, but the composition of glucosinolates shifted, as gluconapin concentrations were reduced, while gluconapoleiferin concentrations increased in plants exposed to nematodes. Aphid presence increased 4-methoxyglucobrassicin concentrations in leaves, which correlated positively with the number of aphids per plant. Nematodes decreased amino acid and sugar concentrations in the phloem. Aphid population doubling time correlated negatively with amino acids and glucosinolate levels in leaves, whereas these correlations were non-significant when nematodes were present. In conclusion, the effects of an herbivore on plant metabolites were independent of the presence of another herbivore. Nematode presence reduced aphid population growth and disturbed feeding relations between plants and aphids. Electronic supplementary material The online version of this article (doi:10.1007/s10886-013-0338-4) contains supplementary material, which is available to authorized users.

Light of different wavelengths affects various physiological processes in plants. Short-wavelength radiation (like UV) can activate defense pathways in plants and enhance the biosynthesis of secondary metabolites (such as flavonoids and glucosinolates) responsible for resistance against certain herbivorous insects. The intensity of light-induced, metabolite-based resistance is plant- and insect species-specific and depends on herbivore feeding guild and specialization. In this study, broccoli (Brassica oleracea var. italica) plants were grown for 4 weeks in a climate chamber under conventional fluorescent tubes and were additionally treated with UV-B (310 nm), UV-A (365 or 385 nm), or violet (420 nm) light generated with UV-B tubes or light-emitting diodes (LEDs). The objective was to determine the influence of narrow bandwidths of light (from UV-B to violet) on plant secondary metabolism and on the performance of the cabbage aphid Brevicoryne brassicae (a specialist) and the green peach aphid Myzus persicae (a generalist). Among flavonol glycosides, specific quercetin and kaempferol glycosides increased markedly under UV-B, while among glucosinolates only 4-methoxy-3-indolylmethyl showed a 2-fold increase in plants exposed to UV-B and UV-A. The concentration of 3-indolylmethyl glucosinolate in broccoli plants increased with UV-B treatment. Brevicoryne brassicae adult weights and fecundity were lower on UV-B treated plants compared to UV-A or violet light-treated plants. Adult weights and fecundity of M. persicae were increased under UV-B and UV-A treatments. When specific light wavelengths are used to induce metabolic changes in plants, the specificity of the induced effects on herbivores should be considered.

Beneficial soil microbes can promote plant growth and induce systemic resistance (ISR) in aboveground tissues against pathogens and herbivorous insects. Despite the increasing interest in microbial-ISR against herbivores, the underlying molecular and chemical mechanisms of this phenomenon remain elusive. Using Arabidopsis thaliana and the rhizobacterium Pseudomonas simiae WCS417r (formerly known as P. fluorescens WCS417r), we here evaluate the role of the JA-regulated MYC2-branch and the JA/ET-regulated ORA59-branch in modulating rhizobacteria-ISR to Mamestra brassicae by combining gene transcriptional, phytochemical, and herbivore performance assays. Our data show a consistent negative effect of rhizobacteria-mediated ISR on the performance of M. brassicae. Functional JA- and ET-signaling pathways are required for this effect, as shown by investigating the knock-out mutants dde2-2 and ein2-1. Additionally, whereas herbivory mainly induces the MYC2-branch, rhizobacterial colonization alone or in combination with herbivore infestation induces the ORA59-branch of the JA signaling pathway. Rhizobacterial colonization enhances the synthesis of camalexin and aliphatic glucosinolates (GLS) compared to the control, while it suppresses the herbivore-induced levels of indole GLS. These changes are associated with modulation of the JA-/ET-signaling pathways. Our data show that the colonization of plant roots by rhizobacteria modulates plant-insect interactions by prioritizing the JA/ET-regulated ORA59-branch over the JA-regulated MYC2-branch. This study elucidates how microbial plant symbionts can modulate the plant immune system to mount an effective defense response against herbivorous plant attackers. Electronic supplementary material The online version of this article (doi:10.1007/s10886-016-0787-7) contains supplementary material, which is available to authorized users.

Glucosinolates are plant secondary defense metabolites confined nearly exclusively to the order Brassicales. Upon tissue rupture, glucosinolates are hydrolyzed to various bioactive breakdown products by the endogenous plant enzyme myrosinase. As the feeding of chewing insect herbivores is associated with plant tissue damage, these insects have developed several independent strategies for coping with the glucosinolate-myrosinase defense system. On the other hand, our knowledge of how phloem-feeding insects interact with the glucosinolate-myrosinase system is much more limited. In fact, phloem feeders might avoid contact with myrosinase altogether so their susceptibility to intoxication by glucosinolate hydrolysis products is unclear. Previous studies utilizing Arabidopsis thaliana plants accumulating high levels of aliphatic- or indolic-glucosinolates indicated that both glucosinolate groups have moderate negative effects on the reproductive performance of Bemisia tabaci, a generalist phloem-feeding insect. To get a deeper understanding of the interaction between B. tabaci and glucosinolate-defended plants, adults were allowed to feed on artificial diet containing intact glucosinolates or on Brussels sprout and A. thaliana plants, and their honeydew was analyzed for the presence of possible metabolites. We found that B. tabaci is capable of cleaving off the sulfate group of intact glucosinolates, producing desulfoglucosinolates that cannot be activated by myrosinases, a mechanism described to date only in several chewing insect herbivores. The presence of desulfated glucosinolates in the honeydew of a generalist phloem-feeder may indicate the necessity to detoxify glucosinolates, likely due to some level of cellular damage during feeding, which results in glucosinolate activation, or as a mechanism to circumvent the non-enzymatic breakdown of indolic glucosinolates.

The performance of natural enemies, such as parasitoid wasps, is affected by differences in the quality of the host’s diet, frequently mediated by species or population-related differences in plant allelochemistry. Here, we compared survival, development time, and body mass in a generalist herbivore, the cabbage moth, Mamestra brassicae, and its solitary endoparasitoid, Microplitis mediator, when reared on two cultivated (CYR and STH) and three wild (KIM, OH, and WIN) populations of cabbage, Brassica oleracea. Plants either were undamaged or induced by feeding of larvae of the cabbage butterfly, Pieris rapae. Development and biomass of M. brassicae and Mi. mediator were similar on both cultivated and one wild cabbage population (KIM), intermediate on the OH population, and significantly lower on the WIN population. Moreover, development was prolonged and biomass was reduced on herbivore-induced plants. However, only the survival of parasitized hosts (and not that of healthy larvae) was affected by induction. Analysis of glucosinolates in leaves of the cabbages revealed higher levels in the wild populations than cultivars, with the highest concentrations in WIN plants. Multivariate statistics revealed a negative correlation between insect performance and total levels of glucosinolates (GS) and levels of 3-butenyl GS. However, GS chemistry could not explain the reduced performance on induced plants since only indole GS concentrations increased in response to herbivory, which did not affect insect performance based on multivariate statistics. This result suggests that, in addition to aliphatic GS, other non- GS chemicals are responsible for the decline in insect performance, and that these chemicals affect the parasitoid more strongly than the host. Remarkably, when developing on WIN plants, the survival of Mi. mediator to adult eclosion was much higher than in its host, M. brassicae. This may be due to the fact that hosts parasitized by Mi. mediator pass through fewer instars, and host growth is arrested when they are only a fraction of the size of healthy caterpillars. Certain aspects of the biology and life-history of the host and parasitoid may determine their response to chemical challenges imposed by the food plant.

The allelopathic potential of the Eurasian invasive plant Alliaria petiolata has been well documented, with the bulk of the effects believed to be mediated by arbuscular mycorrhizal fungi (AMF). We exposed the herbaceous annual Impatiens pallida, which is native to North America, to fractionated A. petiolata extracts at four developmental stages (germination, presymbiosis growth, symbiosis formation, and symbiosis growth) by using exposure levels expected to be similar to field levels. Surprisingly, we found strong direct effects on I. pallida germination and growth, but no indirect effects on I. pallida growth mediated by AMF. We also observed strong synergistic effects with a complete A. petiolata extract that inhibited I. pallida germination and presymbiosis root growth more than either a glucosinolate or flavonoid enriched fraction alone. In fact, the flavonoid enriched fraction tended to stimulate germination and presymbiosis root growth. In contrast to these strong direct effects, I. pallida plant growth during both the symbiosis formation and symbiosis growth phases was unaffected by A. petiolata extracts. We also found no inhibition of AMF colonization of roots or soils by A. petiolata extracts. We show that AMF can actually ameliorate allelopathic effects of an invasive plant, and suggest that previously observed allelopathic effects of A. petiolata may be due to direct inhibition of plant and fungal growth before symbiosis formation.

Invasive plants are by definition excellent competitors, either indirectly through competition for resources or directly through allelopathic inhibition of neighboring plants. Although both forms of competition are commonly studied, attempts to explore the interactions between direct and indirect competition are rare. We monitored the effects of several doses of extracts of Alliaria petiolata, a Eurasian invader in North America, on the growth of Impatiens pallida, a North American native, at several planting densities. The density-dependent phytotoxicity model predicts that as plant density increases, individual plant size will decrease, unless a toxin is present in the soil. In this case, individual plant size is predicted to increase as plant density increases, as plants share a limited toxin dose. We tested this model using fractions of an A. petiolata extract enriched in flavonoids or glucosinolates, as well as a combined fraction. The flavonoid-enriched fraction and the combined fraction suppressed I. pallida growth but only when applied at a dose eight times higher than that expected in the field. When treated with a dose equivalent to estimated field exposure levels, I. pallida growth was not distinguishable from that of control plants that received no extract, showing that indirect competition for resources was more important for determining the growth of I. pallida than direct allelopathic inhibition by A. petiolata. This is an important reminder that, even though many plants have the demonstrated potential to exert strong allelopathic effects, those effects may not always be apparent when other forms of competition are considered as well.

The effects of plant competition and herbivory on glucosinolate concentrations in cabbage root and foliage were investigated in a cabbage-red clover intercropping system. Cabbage plants were grown under different competitive pressures and with varying degrees of attack by root-feeding Delia floralis larvae. Glucosinolate concentrations in cabbage were affected both by intercropping and by D. floralis density. Glucosinolate concentrations in foliage generally decreased as a response to intercropping, while the responses to insect root damage of individual glucosinolates were weaker. Root glucosinolates responded more strongly to both intercropping and egg density. Total root glucosinolate concentration decreased with clover density, but only at high egg densities. Increased egg density led to opposite reactions by the indole and aliphatic glucosinolates in roots. The responses of individual root glucosinolates to competition and root damage were complex and, on occasion, nonlinear. Reduced concentrations of several glucosinolates and the tendency towards a decrease in total concentration in cabbage foliage caused by intercropping and larval damage suggest that competing plants or plants with root herbivory do not allocate the same resources as unchallenged plants towards sustaining levels of leaf defensive compounds. This could also be true for root glucosinolate concentrations at high egg densities. In addition, the results suggest that changes occurring within a structural group of glucosinolates may be influenced by changes in a single compound, e.g., glucobrassicin (indol-3-ylmethyl) in foliage or sinigrin (2-propenyl) in roots.

Thlaspi praecox Wulfen (Brassicaceae) is a perennial Cd/Zn hyperaccumulating plant species that forms functional arbuscular mycorrhizal (AM) symbiosis. Glucosinolates (GS) were studied in different organs of field-collected T. praecox at differing plant developmental stages. Additionally, AM colonization was recorded. Total GS concentrations and profiles of nine individual GS varied during the plant life cycle. Novel individual GS that were related to specific developmental phases, mainly to flowering and seed production, were identified. The highest total GS and sinalbin concentrations in rosette leaves were found in the vegetative phase, possibly contributing to protection of young, palatable leaves. The lowest were found in roots during the flowering and the seeding phases. Increased total GS concentrations in roots and enhanced aliphatic GS, especially glucobrassicanapin, in the senescence phase may protect roots from herbivory during winter and early spring. The presence of glucotropaeolin and the absence of glucobrassicanapin in the flowering phase coincided with peak AM colonization. This is the first report on GS profiles in an AM and metal-hyperaccumulating plant.

The composition of secondary metabolites and the nutritional value of a plant both determine herbivore preference and performance. The genetically determined glucosinolate pattern of Barbarea vulgaris can be dominated by either glucobarbarin (BAR-type) or by gluconasturtiin (NAS-type). Because of the structural differences, these glucosinolates may have different effects on herbivores. We compared the two Barbarea chemotypes with regards to the preference and performance of two lepidopteran herbivores, using Mamestra brassicae as a generalist and Pieris rapae as a specialist. The generalist and specialist herbivores did not prefer either chemotype for oviposition. However, larvae of the generalist M. brassicae preferred to feed and performed best on NAS-type plants. On NAS-type plants, 100% of the M. brassicae larvae survived while growing exponentially, whereas on BAR-type plants, M. brassicae larvae showed little growth and a mortality of 37.5%. In contrast to M. brassicae, the larval preference and performance of the specialist P. rapae was unaffected by plant chemotype. Total levels of glucosinolates, water soluble sugars, and amino acids of B. vulgaris could not explain the poor preference and performance of M. brassicae on BAR-type plants. Our results suggest that difference in glucosinolate chemical structure is responsible for the differential effects of the B. vulgaris chemotypes on the generalist herbivore. Electronic supplementary material The online version of this article (doi:10.1007/s10886-007-9424-9) contains supplementary material, which is available to authorized users.

Through artificial selection, domesticated plants often contain modified levels of primary and secondary metabolites compared to their wild progenitors. It is hypothesized that the changed chemistry of cultivated plants will affect the performance of insects associated with these plants. In this paper, the development of several specialist and generalist herbivores and their endoparasitoids were compared when reared on a wild and cultivated population of cabbage, Brassica oleracea, and a recently established feral Brassica species. Irrespective of insect species or the degree of dietary specialization, herbivores and parasitoids developed most poorly on the wild population. For the specialists, plant population influenced only development time and adult body mass, whereas for the generalists, plant populations also affected egg-to-adult survival. Two parasitoid species, a generalist (Diadegma fenestrale) and a specialist (D. semiclausum), were reared from the same host (Plutella xylostella). Performance of D. semiclausum was closely linked to that of its host, whereas the correlation between survival of D. fenestrale and host performance was less clear. Plants in the Brassicaceae characteristically produce defense-related glucosinolates (GS). Levels of GS in leaves of undamaged plants were significantly higher in plants from the wild population than from the domesticated populations. Moreover, total GS concentrations increased significantly in wild plants after herbivory, but not in domesticated or feral plants. The results of this study reveal that a cabbage cultivar and plants from a wild cabbage population exhibit significant differences in quality in terms of their effects on the growth and development of insect herbivores and their natural enemies. Although cultivated plants have proved to be model systems in agroecology, we argue that some caution should be applied to evolutionary explanations derived from studies on domesticated plants, unless some knowledge exists on the history of the system under investigation.

The glucosinolate-myrosinase system of Brassicaceae is known to hold a defensive function in both a constitutive and an inducible fashion. Glucosinolates are sulfur- and nitrogen-containing metabolites that are hydrolyzed upon tissue disruption by myrosinase enzymes. The resulting products are toxic for most herbivores. Nevertheless, some insects evolved detoxification mechanisms that enable them to feed exclusively on Brassicaceae. Induction of plant chemical defenses that deter or poison generalists might be ineffective against adapted specialists. We investigated the specificity of short-term induction patterns of chemical defenses in Sinapis alba damaged by a glucosinolate-sequestering specialist herbivore (turnip sawfly, Athalia rosae), a generalist herbivore (fall armyworm, Spodoptera frugiperda), or mechanical wounding (cork borer), and their effects on the behavior of A. rosae. After 24 hr of damage to young leaves, local as well as systemic changes in glucosinolate and myrosinase levels were analyzed. The intensity of the resulting changes was highest in damaged leaves. Induction responses in S. alba were dependent upon the attacking herbivore and were distinct from a mere wound response. Specialist feeding and mechanical wounding evoked up to threefold increases in levels of both parts of the glucosinolate-myrosinase system, whereas generalist feeding induced up to twofold increases in glucosinolate levels only. The majority of constitutive and induced myrosinase activity was found in the insoluble fractions. Possible consequences for the plant-specialist interaction were examined in behavioral tests with larvae and adult females of A. rosae on induced S. alba plants. Larval feeding and adult oviposition patterns were not modulated in relation to plant treatment. Thus, specificity was found in S. alba responses in relation to the inducing agent, but it was not present in return in the effects on the behavior of an adapted herbivore.

As it pertains to insect herbivores, the preference-performance hypothesis posits that females will choose oviposition sites that maximize their offspring's fitness. However, both genetic and environmental cues contribute to oviposition preference, and occasionally "oviposition mistakes" occur, where insects oviposit on hosts unsuitable for larval development. Pieris virginiensis is a pierine butterfly native to North America that regularly oviposits on an invasive plant, Alliaria petiolata, but the caterpillars are unable to survive. Alliaria petiolata has high concentrations of the glucosinolate sinigrin in its tissues, as well as a hydroxynitrile glucoside, alliarinoside. We investigated sinigrin as a possible cause of mistake oviposition, and sinigrin and alliarinoside as possible causes of larval mortality. We found that sinigrin applied to leaves of Cardamine diphylla, a major host of P. virginiensis that does not produce sinigrin, had no effect on oviposition rates. We tested the effect of sinigrin on larval performance using two host plants, one lacking sinigrin (C. diphylla) and one with sinigrin naturally present (Brassica juncea). We found no effect of sinigrin application on survival of caterpillars fed C. diphylla, but sinigrin delayed pupation and decreased pupal weight. On B. juncea, sinigrin decreased survival, consumption, and caterpillar growth. We also tested the response of P. virginiensis caterpillars to alliarinoside, a compound unique to A. petiolata, which was applied to B. oleracea. We found a significant reduction in survival, leaf consumption, and caterpillar size when alliarinoside was consumed. The 'novel weapon' alliarinoside likely is largely responsible for larval failure on the novel host A. petiolata. Sinigrin most likely contributes to the larval mortality observed, however, we did not observe any effect of sinigrin on oviposition by P. virginiensis females. Further research needs to be done on non-glucosinolate contact cues, and volatile signals that may induce P. virginiensis oviposition.

Glucosinolates (GS) and trichomes contribute to plant resistance against insect herbivores in the model Arabidopsis thaliana. The functional and genetic characteristics of herbivore defense, however, can differ even between closely related species. In a quantitative genetic experiment with the out-crossing perennial Arabidopsis lyrata spp. petraea, we measured constitutive GS composition, trichome density, leaf thickness, and plant resistance in four different herbivore interactions. In a single population of A. lyrata, we found heritable variation for trichome density as well as GS amount and carbon side-chain elongation ratios associated with activity in methylthioalkylmalate synthase (MAM). Unexpectedly, heritabilities for indole GS in A. lyrata were high and less affected by differences in plant age and environment than aliphatic GS. We found significant heritability in plant resistance to the specialist Plutella xylostella and generalist Trichoplusia ni, but not to the specialists Pieris brassicae and Phyllotreta cruciferae. Analyses of phenotypic and genetic correlations between candidate defense traits and insect resistance suggested that A. lyrata resistance was conferred by a combination of indole GS amount and trichome density, and, to a lesser extent, aliphatic GS ratios and leaf thickness. Variation in the most abundant compound, the aliphatic 3-hydroxypropyl GS, had little impact on A. lyrata herbivore resistance. The contribution of defense traits to resistance depended on the experimental herbivory context, and resistances were weakly correlated. A diversified defense strategy is likely to be important for long-lived individuals of A. lyrata that are subject to attack by many different herbivores in nature.

The Evolution of Increased Competitive Ability (EICA) hypothesis posits that invasive plants in introduced habitats with reduced herbivore pressure will evolve reduced levels of costly resistance traits. In light of this hypothesis, we examined the constitutive and inducible expression of five chemical defense traits in Alliaria petiolata from four invasive North American and seven native European populations. When grown under common conditions, significant variation among populations within continents was found for trypsin inhibitors and peroxidase activity, and glucosinolates and trypsin inhibitors were significantly jasmonate-inducible across populations. Across populations, constitutive levels of glucosinolates and trypsin inhibitors were negatively correlated with their degree of induction, with three North American populations tending to have lower constitutive levels and higher inducibility of glucosinolates than the seven European populations. Alliarinoside and isovitexin 6"-O-beta-glucopyranoside levels were both higher in North American plants than in European plants, but levels of these compounds were generally increased by jasmonate in European plants and decreased by the same treatment in North American plants. Aside from the tendency for invasive populations to have reduced constitutive glucosinolate levels coupled with increased inducibility, little support for the predictions of EICA was evident in the chemical defenses that we studied.

The interactions of plants with herbivores and pathogens have been suggested to drive the evolution of resistances in plants and in some cases new lineages and taxa. However, such divergence may require reproductive isolation, e.g., in allopatry. In the crucifer Barbarea vulgaris, some plants are resistant to the flea beetle Phyllotreta nemorum, due to production of specific saponins, whereas others are susceptible. Resistant and susceptible plants additionally differ in resistance to the pathogen Albugo candida, content of glucosinolates, and leaf pubescence, and they are genetically strongly divergent and partially reproductively incompatible. This suggests that at some point they were separated for a considerable length of time. Previously, the insect susceptible P-type had been described only from Denmark, Sweden, and Estonia, whereas the resistant G-type is widely distributed in Western Europe. Here, we tested whether the two plant types have divergent geographical distributions and maintain their distinct trait associations throughout their range. The insect-susceptible type was found in Russia, the Baltics, and parts of Fennoscandia, but not in Central Europe. In contrast, the insect resistant type was found from Finland and westwards. Their different trait associations were consistent within the two ranges. We therefore suggest that the two plant types diverged in allopatry at some time in the past, and evolved different resistances in response to local antagonists. The two plant types probably maintain their distinctness due to a hybridization barrier. Thus, the present distributions of the two types may be shaped by both historical processes and current differential biotic selection.

In this study, the performance and sequestration specificity of the sawfly Athalia rosae L. (Hymenoptera: Tenthredinidae) reared on two wild Brassica oleracea L. (Brassicaceae) populations (KIM and WIN), which express indole glucosinolates (GSs) in very high concentrations, were investigated. The local and systemic plant responses to larval feeding also were analyzed. Indole GSs predominated in KIM plants, whereas aliphatic GSs prevailed in the WIN population, which had several times higher total GS concentrations than KIM plants. Plants of the KIM population had lower specific leaf areas, and higher carbon/nitrogen ratios than WIN plants. The insects reared on WIN plants performed better for most traits than insects reared on the KIM population. The larvae preferentially sequestered aliphatic GSs but when feeding on KIM plants, indole GSs were also concentrated in the hemolymph. In response to feeding by A. rosae larvae, indole GSs were induced locally and systemically, whereas aliphatic GSs were reduced only in systemic leaves of the WIN population. Soluble myrosinase activities were constitutively higher in WIN than in KIM plants, and feeding damage led to a significant reduction of this glucosinolate-degrading enzyme in WIN plants only. The data suggest that the sawfly is well adapted to high concentrations of aliphatic GSs, which are sequestered by the larvae. In contrast, the larvae may be less adapted to plants dominated by indole GSs. Selective induction of indole GSs by the plants in response to larval feeding may be adaptive as accumulation of indole GSs may lower the performance of the larvae.

In an attempt to identify chemical signals governing the general flower and silique feeding behavior of larvae of the orange tip butterfly, Anthocharis cardamines (L.), we investigated feeding behavior and chemistry of two major host plants: Cardamine pratensis L. and Alliaria petiolata (Bieb.) Cavara & Grande (garlic mustard). Larvae reportedly feed mainly on flowers and siliques rather than leaves in nature, and did so when observed on the original host plants. Behavioral experiments, using detached A. petiolata branches, however, showed that larvae readily accepted leaves and only the final instar showed a tendency for directed movement towards floral parts. To search for semiochemicals that control plant part preference and to assess possible nutritional consequences of floral parts feeding, we determined glucosinolate profiles and total nitrogen levels of floral parts and leaves. There was only moderate difference between glucosinolate profiles of leaves and floral parts within each of two host plant species. In contrast, the profiles of floral parts differed significantly between them. A. petiolata was dominated by 2-propenyl glucosinolate, while C. pratensis was dominated by aromatic glucosinolates and branched aliphatic glucosinolates, with considerable variation among populations. Nitrogen levels tended to be higher in floral parts than in leaves in A. petiolata, but not in C. pratensis, so floral feeding could not generally be attributed to higher N content. With the exception of a tendency of last instar larvae (L5) to move to the apex and ingest flowers and upper stem, we did not find either a plant chemistry basis or larval acceptance/rejection behavior that could explain the usual feeding of floral parts by orange tip larvae of all instars. However, by artificial manipulation of vertical larval position on host plants, we found that the frequency of leaf vs. flower feeding during 24 hr depended significantly on the initial larval position. Hence, we suggest that the placement of eggs on floral parts by ovipositing female butterflies is a major explanation of orange tip feeding habits previously known from field observations.

Glucosinolates are a diverse group of defensive secondary metabolites that is characteristic of the Brassicales. Arabidopsis thaliana (L.) Heynh. (Brassicaceae) lines with mutations that greatly reduce abundance of indole glucosinolates (cyp79B2 cyp79B3), aliphatic glucosinolates (myb28 myb29), or both (cyp79B2 cyp79B3 myb28 myb29) make it possible to test the in vivo defensive function of these two major glucosinolate classes. In experiments with Lepidoptera that are not crucifer-feeding specialists, aliphatic and indole glucosinolates had an additive effect on Spodoptera exigua (Hubner) (Lepidoptera: Noctuidae) larval growth, whereas Trichoplusia ni (Hubner) (Lepidoptera: Noctuidae) and Manduca sexta (L.) (Lepidoptera: Sphingidae) were affected only by the absence of aliphatic glucosinolates. In the case of two crucifer-feeding specialists, Pieris rapae (L.) (Lepidoptera: Pieridae) and Plutella xylostella (L.) (Lepidoptera: Plutellidae), there were no major changes in larval performance due to decreased aliphatic and/or indole glucosinolate content. Nevertheless, choice tests show that aliphatic and indole glucosinolates act in an additive manner to promote larval feeding of both species and P. rapae oviposition. Together, these results support the hypothesis that a diversity of glucosinolates is required to limit the growth of multiple insect herbivores.

The diamondback moth (Plutella xylostella), a crucifer-specialist pest, has been documented to employ glucosinolates as host recognition cues for oviposition. Through the use of mutant Arabidopsis thaliana plants, we investigated the role of specific classes of glucosinolates in the signaling of oviposition by P. xylostella in vivo. Indole glucosinolate production in A. thaliana was found to be crucial in attracting oviposition. Additionally, indole glucosinolates functioned as oviposition cues only when in their intact form. 4-Methoxy-indol-3-ylmethylglucosinolate was implicated as an especially strong oviposition attractant in vitro, suggesting that indole glucosinolate secondary structure may play a role in P. xylostella host recognition as well. Aliphatic glucosinolate-derived breakdown products were found to attract P. xylostella, but only after damage or in the absence of indole glucosinolates. Furthermore, mutant plants lacking both intact indole glucosinolates and aliphatic glucosinolate breakdown products exhibited decreased oviposition attractiveness beyond that of the progenitor mutants lacking either component of the glucosinolate-myrosinase system. Therefore, we conclude that nonvolatile indole glucosinolates and volatile aliphatic glucosinolate breakdown products both appear to play important roles as host recognition cues for P. xylostella oviposition.

Cruciferous plants (Brassicaceae) are characterized by the accumulation of a group of secondary metabolites known as glucosinolates that, following attack by pathogens or herbivores, may be hydrolyzed to one of a number of products including isothiocyanates and nitriles. Despite the range of hydrolysis products that may be produced, the toxicity of glucosinolates to pathogens and herbivores may be explained largely by the production of isothiocyanates. Isothiocyanates are also known to provide an indirect defense by acting as host finding cues for parasitoids of insect herbivores that attack crucifers. It has been speculated that nitriles may provide a similar indirect defense. Here, we investigate the olfactory perception and orientation behavior of the aphid parasitoid Diaeretiella rapae, to a range of alkenylglucosinolate hydrolysis products, including isothiocyanates, nitriles, and epithionitriles. Electroantennogram responses indicated peripheral odor perception in D. rapae females to all 3-butenylglucosinolate hydrolysis products tested. By contrast, of the 2-propenylglucosinolate hydrolysis products tested, only the isothiocyanate elicited significant responses. Despite showing peripheral olfactory detection of a range of 3-butenylglucosinolate hydrolysis products, naive females oriented only to the isothiocyanate. Similarly, parasitoids oriented to 3-isothiocyanatoprop-1-ene, but not to the corresponding nitrile or epithionitrile. However, by rearing D. rapae either on Brassica nigra, characterized by the accumulation of 2-propenylglucosinolate, or Brassica rapa var rapifera, characterized by the accumulation of 3-butenylglucosinolate, altered the innate response of parasitoids to 3-isothiocyanatoprop-1-ene and 4-isothiocyanatobut-1-ene. These results are discussed in relation to the defensive roles of glucosinolate hydrolysis products and the influence of the host plant on aphid parasitoid behavior.

Glucosinolates are commonly found in Arabidopsis thaliana and its crucifer relatives, which are known for their role in defense against insect herbivory. In a common garden experiment, we assessed genotypic variation in glucosinolates in A. thaliana and evaluated the association between this chemistry and both plant damage and fitness. Specifically, glucosinolate concentrations were directly associated with damage levels and inversely associated with fitness. These results are contrary to the general expectation that enhanced chemical defense should result in decreased insect herbivory. As the measured insect community in this field trial was dominated by specialist herbivores, this positive relationship between glucosinolates and herbivory agrees with previous observations that glucosinolates (or their hydrolysis products) attract specialist insects. In addition, glucosinolate diversity in this common garden appeared to affect herbivore damage levels. For example, genotypes that contained alkenyl glucosinolates had higher mean damage levels than those that contained hydroxyalkyl glucosinolates. Results suggest that genotypic variation in glucosinolates may be a major factor in determining plant utilization patterns by insect herbivores in the field.

Brassica nigra plants, characterized by high levels of sinigrin, and artificial aphid diets to which sinigrin was selectively added were used to rear the crucifer specialist, Brevicoryne brassicae. Aphids were provided as a food source to two species of polyphagous ladybird, Adalia bipunctata and Coccinella septempunctata. First instar A. bipunctata were unable to survive when fed with B. brassicae reared on B. nigra or diets containing 0.2% sinigrin, but when fed with aphids reared on diets containing 0% sinigrin, survival rates were high. By contrast, first instar C. septempunctata were able to survive when fed with aphids reared on B. nigra or artificial diets containing up to 1% sinigrin. However, the presence of sinigrin in the aphid diet decreased larval growth and increased the time necessary for larvae to reach second instar for this species of ladybird. These results indicate that the presence of sinigrin in the diet of B. brassicae makes this aphid unsuitable as a food source for A. bipunctata but not for C. septempunctata, although for this ladybird species, there appear to be costs associated with feeding on aphids that contain this secondary metabolite.

Desert locusts (Schistocerca gregaria) occasionally feed on Schouwia purpurea, a plant that contains tenfold higher levels of glucosinolates than most other Brassicaceae. Whereas this unusually high level of glucosinolates is expected to be toxic and/or deterrent to most insects, locusts feed on the plant with no apparent ill effects. In this paper, we demonstrate that the desert locust, like larvae of the diamondback moth (Plutella xylostella), possesses a glucosinolate sulfatase in the gut that hydrolyzes glucosinolates to their corresponding desulfonated forms. These are no longer susceptible to cleavage by myrosinase, thus eliminating the formation of toxic glucosinolate hydrolysis products. Sulfatase is found throughout the desert locust gut and can catalyze the hydrolysis of all of the glucosinolates present in S. purpurea. The enzyme was detected in all larval stages of locusts as well as in both male and female adults feeding on this plant species. Glucosinolate sulfatase activity is induced tenfold when locusts are fed S. purpurea after being maintained on a glucosinolate-free diet, and activity declines when glucosinolates are removed from the locust diet. A detoxification system that is sensitive to the dietary levels of a plant toxin may minimize the physiological costs of toxin processing, especially for a generalist insect herbivore that encounters large variations in plant defense metabolites while feeding on different species.

Interactions between butterflies and caterpillars in the genus Pieris and plants in the family Brassicaceae are among the best explored in the field of insect-plant biology. However, we report here for the first time that Pieris brassicae, commonly assumed to be a typical folivore, actually prefers to feed on flowers of three Brassica nigra genotypes rather than on their leaves. First- and second-instar caterpillars were observed to feed primarily on leaves, whereas late second and early third instars migrated via the small leaves of the flower branches to the flower buds and flowers. Once flower feeding began, no further leaf feeding was observed. We investigated growth rates of caterpillars having access exclusively to either leaves of flowering plants or flowers. In addition, we analyzed glucosinolate concentrations in leaves and flowers. Late-second- and early-third-instar P. brassicae caterpillars moved upward into the inflorescences of B. nigra and fed on buds and flowers until the end of the final (fifth) instar, after which they entered into the wandering stage, leaving the plant in search of a pupation site. Flower feeding sustained a significantly higher growth rate than leaf feeding. Flowers contained levels of glucosinolates up to five times higher than those of leaves. Five glucosinolates were identified: the aliphatic sinigrin, the aromatic phenylethylglucosinolate, and three indole glucosinolates: glucobrassicin, 4-methoxyglucobrassicin, and 4-hydroxyglucobrassicin. Tissue type and genotype were the most important factors affecting levels of identified glucosinolates. Sinigrin was by far the most abundant compound in all three genotypes. Sinigrin, 4-hydroxyglucobrassicin, and phenylethylglucosinolate were present at significantly higher levels in flowers than in leaves. In response to caterpillar feeding, sinigrin levels in both leaves and flowers were significantly higher than in undamaged plants, whereas 4-hydroxyglucobrassicin leaf levels were lower. Our results show that feeding on flower tissues, containing higher concentrations of glucosinolates, provides P. brassicae with a nutritional benefit in terms of higher growth rate. This preference appears to be in contrast to published negative effects of volatile glucosinolate breakdown products on the closely related Pieris rapae.

To examine how plant allelochemicals in prey affect foraging choices made by generalist predator paper wasps, Polistes dominulus (Vespidae), we compared predation on Pieris napi (Pieridae) caterpillars reared on host plants with different allelochemicals. In naturalistic behavioral choice experiments, free-flying wasps chose between living pierids reared on cabbage (Brassica oleracea), which lacks deterrent allelochemicals, or alternate host plants with potentially deterrent allelochemicals. The alternative host plants were: wormwood mustard, (Erysimum cheiranthoides: Brassicaceae), which contains cardenolides; nasturtium (Tropaeolum majus: Tropaeolaceae) with high concentrations of chlorogenic acid; and black mustard (Brassica nigra: Brassicaceae) with high concentrations of the aliphatic glucosinolate, sinigrin. Although wasps captured equal numbers of caterpillars reared on cabbage and each alternate host plant, they spent significantly longer handling prey from the alternate host plants as they selectively removed the caterpillar's gut, which contained the plant material. This was true even if the caterpillar did not sequester toxins in its tissues, as revealed by high performance liquid chromatography (HPLC) analysis of Erysimum-reared pierids. Because handling time is longer, predators that capture pierids containing non-sequestered allelochemicals experience an overall reduction in foraging rate that may translate into a fitness cost.

Variation in chemical defenses remains underexplored. In particular, little is known about patterns of variation at small scales within leaves and spatial variation of induction. I examined variation in glucosinolate concentrations in the leaves of Raphanus sativus at several different spatial scales in two related experiments. I used samples equivalent in area to the amount an intermediate-sized caterpillar might eat in 1 d, a smaller scale than used in most previous studies. I examined variation due to induction and leaf age and small-scale spatial variation within leaves. The mean and variance of glucosinolate concentrations were higher in induced plants, young leaves, and the proximal half of leaves. Higher glucosinolate concentrations in the proximal half of leaves are previously unreported. Small-scale variation was extreme, accounting for 57% of the total random variation, and spatially random. There was no spatial autocorrelation found at scales as small as 1-2 cm. The high degree of small-scale, spatially random variation in glucosinolate concentrations in leaves is previously unreported. This small-scale variation and the variation caused by induction may have significant effects on herbivores and could be an important component of plant defense.