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Insect pest occurrences on cabbage fields intercropped with leafy daikon, which is known to present a decoy plant for clubroot disease and belongs to the same family as cabbage (Brassicaceae), were examined at open field in the Miyagi prefecture, northern Japan. Four experiments were conducted to examine the influence of: intercropped leafy daikon’s seeding density on insect pest occurrence, trimming of intercropped leafy daikon on both insect pest occurrence and cabbage yield, and mix-cropping of leafy daikon and cabbage on insect pest occurrence. Leafy daikon intercropping reduced the occurrence of some insect pest species, especially that of the pests categorized as leaf-chewing pests, such as beet semi-loopers, Autographa nigrisigna Walker (Lepidoptera: Noctuidae), and small white butterflies, Pieris rapae crucivora Boisduval (Lepidoptera: Pieridae), and the results were compared with their occurrences in barley intercropping and non-intercropping. These two insect species were rarely observed on intercropped and mix-cropped leafy daikon, indicating it did not work as a trap crop. However, a reduction in cabbage yield due to shading of leafy daikon intercropping was observed, but the yield loss was mitigated by trimming of intercropped leafy daikon during the cultivation. Furthermore, leafy daikon intercropping with trimming had the same occurrence reduction effects against A. nigrisigna and P. rapae crucivora as leafy daikon intercropping without trimming. Our findings suggested that intercropping of closely related plants was possible by selecting the appropriate combinations of parameters and proper strategies, which proved to be useful for insect pest management in mix-cropping systems.

Plants grow under reduced water availability can have divergent effects on insect herbivores, in some instances producing benefits to them. However, the forces mediating these positive impacts remain mostly unclear. We conducted a manipulative field study using a specialist herbivore Pieris rapae, and its host plant, Rorippa indica, in two populations to identify how water availability impacts overall plant quality and multitrophic interactions. We observed that R. indica growing under low water availability led to higher survival of P. rapae larvae. The increase in survival of eggs and larvae was related to the reduced abundance of other herbivores and natural enemies. Water availability had differential impacts on members of the herbivore community through changes in plant quality. Low water availability decreased the quality of R. indica to most herbivores as indicated by reduced abundance in the field and decreased relative growth rate in feeding assays. In contrast, the performance of P. rapae larvae were not affected by differences in sympatric R. indica grown under different water availability. These results indicate that local P. rapae possess some physiological adaptation to overcome fluctuations in host quality. Our findings illustrate that reduced water availability is beneficial to a specialist herbivore, but detrimental to most other herbivores. Our work highlights the complex roles of the arthropod communities associated with plants in determining the impacts of water availability on insect herbivores.

Insect population count and yield were used to determine the effectiveness of two bio-rational pesticides -a commercial formulation of Pieris rapae granulosis virus and Bacillus thuriengensis, Bypel 1® (PrGV + Bt) (1.5 g/l w/v) and aqueous neem kernel extract (ANKE) (50 g/l w/v), on the insect pests complex of two cabbage varieties, KK cross and Oxylus for two consecutive cropping seasons of 2016/2017. The experiment was laid in a split-plot design with three replications, with the cabbage varieties as main plots and the bio-rational pesticides as subplots. Cabbage plots treated with Bypel 1® and ANKE had the lowest population of Plutella xylostella, Brevicoryne brassicae, Hellula undalis, Bemisia tabaci and Thrips tabaci for both seasons. The control plot consistently had higher pest population in both seasons. There was no detrimental effect of bio-rational pesticides on natural enemies. The bio-rational pesticides-treated plots produced higher yield than control plots. Mean numbers of P. xylostella was higher on KK (0.61, 1.46) compared to Oxylus (0.65, 1.70) in both seasons, respectively. The yield between the cabbage varieties was not significantly different in both seasons, although Bypel 1® and ANKE treated plots generally produced significantly higher yields than the control plot. Yield between both cropping seasons was significantly different (p = 0.039). These findings provide evidence that bio-rational pesticides may offer effective management of the pest complex of cabbage and may be successfully used as an integral part of IPM in Africa as a means to curb the abuse of synthetic insecticides.

Interspecific competition for limited resources can drive ecological specialization and trait expression. Organisms released from intense competition may exploit a broader range of resources, but if reunited with stronger competitors, survivorship may depend on foraging behaviors that reduce competition. We compared the host selection behavior of the parasitoid Cotesia glomerata from two North American populations that differ in their association with Cotesia rubecula, a superior competitor. Both parasitoids originate from Europe and attack the imported cabbageworm (a.k.a. small cabbage white) Pieris rapae, but C. glomerata was introduced into North America almost a century before C. rubecula. After re-association in North America, C. rubecula has displaced C. glomerata in several regions, but not in other regions. Host selection was measured in female C. glomerata from Maryland (MD) where it coexists with C. rubecula, and in conspecifics from Colorado (CO) where C. rubecula is absent. Unparasitized and C. rubecula-parasitized P. rapae hosts were used in choice tests to examine whether C. glomerata host selection behavior differed based on the population’s association history with C. rubecula. We found that C. glomerata from MD had a higher likelihood of avoiding hosts parasitized by C. rubecula (and thus avoiding competition) than did wasps from CO. The ability of C. glomerata to avoid hosts parasitized by C. rubecula may facilitate coexistence in MD; whereas, the lack of discrimination in CO populations of C. glomerata naïve to C. rubecula could contribute to the displacement of C. glomerata were C. rubecula to enter the same habitat.

The two subspecies of the small white butterfly, the European Pieris rapae rapae and the Asian P. r. crucivora, differ in wing colouration. Under ultraviolet light, the wings of both male and female P. r. rapae appear dark, whereas the wings of male P. r. crucivora are dark and those of females are bright. It has been hypothesized that these sexually dimorphic wing reflections in P. r. crucivora may have induced the evolution of a fluorescing-screening pigment in the violet-opsin-expressing photoreceptors of males, thus facilitating greater wavelength discrimination near 400nm. Comparing the compound eyes of the two subspecies using genetic, microscopical, spectrographic, and histological methods revealed no differences that would meaningfully affect photoreceptor sensitivity, suggesting that the fluorescing-screening pigment did not evolve in response to sexually dimorphic wing reflections. Our investigation further revealed that (i) the peri-rhabdomal reddish-screening pigments differ among the three ommatidial types; (ii) each of the ommatidial types exhibits a unique class of red photoreceptor with a distinct spectral peak; and (iii) the blue, green, and red photoreceptors of P. rapae exhibit a polarization sensitivity >2, with red photoreceptors allowing for a two-channel opponency form of polarization sensitivity.

Oligophagous insects can consume a wide range of different host plant species, but how these host plants vary in their metabolite compositions and the extent to which this variation affects the biochemistry of the insect herbivores is largely unknown. An understanding of how defensive metabolites from plants are processed by insects may help us develop more effective pesticides. We studied the interactions between the oligophagous insect herbivore Pieris rapae (Lepidoptera: Pieridae) and five species of its larval host plants (family Brassicaceae and Cleomaceae) by examining untargeted metabolic fingerprints of the plants and the larval herbivores feeding on them. Visualisation of the metabolic fingerprints of the different host plant species showed highly distinctive clusters in the PCA-X score plots. Larvae could also be distinguished based on the species of host plant they fed on but clusters overlapped to a greater extent. The fingerprints of larvae feeding on Cleome spinosa plants were most distinctive due to a large group of abundant metabolites also found in high abundance in C. spinosa, but not in the other host plants examined. We conclude that host plants influence the biochemistry of their larval herbivores, and that some metabolites are conserved from one trophic level to the next.

Mating might significantly affect the host selection behaviors of phytophagous insects. Here, we investigated the post-mating changes in behavioral and antennal responses of Pieris rapae (Lepidoptera: Pieridae) females to host plant volatiles. In two-choice bioassays using artificial plant models, mated females visited the model scented with synthetic blends (15-, 9-, or 6-components) of cabbage plant volatiles more frequently than the unscented control, whereas virgin females did not exhibit this preference. Because single compounds and the 3-component blend did not induce preferential visiting, mated females apparently utilized complex odor blends as their host-finding cue. Moreover, 2- to 4-day-old mated females visited the models, scented and unscented, more frequently than did their virgin counterparts. Therefore, mating enhanced the host-finding behavior of young females and their responsiveness to plant volatiles. Gas chromatography-electroantennographic detector analysis revealed that eight of the 15 compounds in the cabbage plant volatiles elicited responses from female antennae. However, post-mating and age-dependent changes in antennal responses were not detected. Because female peripheral (antennal) sensitivity to volatiles remained practically unchanged after emergence, post-mating changes in host selection might be attributed to changes in the central nervous system.

To increase the tolerance of Chinese cabbage (Brassica campestris L. ssp. pekinensis) to Pieris rapae, we introduced a modified cowpea trypsin inhibitor (CpTI) gene, SCK, into various cultivars. SCK was derived from CpTI, an insect-resistance gene. The protein accumulating capacity of CpTI can be improved by adding a signal peptide sequence at the 5′ end and an endoplasmic reticulum-detained signal sequence at the 3′ end. Using an optimized Agrobacterium tumefaciens-mediated transformation system in Chinese cabbage, we obtained a maximum transformation efficiency of ~6.83%. Insect resistance tests and CpTI enzymatic assays showed that most of the transgenic plants had significant resistance to cabbage worm (Pieris rapae) larvae and that the plants with the highest levels of insect resistance had the greatest CpTI-related capacity, indicating a high correlation between SCK expression and insect resistance. An evaluation of segregation patterns in the independent transgenic line with the highest insect resistance, ‘ZB-08-04’, showed that kanamycin resistant versus sensitive plants segregated in a 3:1 Mendelian fashion. This study provides a potential germplasm resource for Chinese cabbage breeding in the future.

Butterflies are regularly used as model systems for understanding the role of coloration in communication because of their highly variable and conspicuous phenotypes. Most research showing a role for color in communication has focused on aspects of brightness or hue of entire wings or large color patches. However, evidence is accumulating that butterflies sometimes use smaller wing pattern elements in communication. Here we provide evidence that both male and female cabbage white butterflies (Pieris rapae L.) discriminate among conspecifics on the basis of the number of small but conspicuous black wing spots of the dorsal forewing. Male butterflies were more interactive with model butterflies with two spots, which resemble female butterflies, than with model butterflies with only one spot, which resemble male butterflies. Female butterflies showed the opposite response, being more interactive with male-like (one-spot) models than with female-like (two-spot) models. Some of our experiments were conducted with an electronic device designed to create a realistic and controlled fluttering behavior of the models. We describe the design and function of this device and provide evidence that it increased butterfly responses compared to a non-fluttering model. This device could prove useful for others addressing questions of communication in butterflies or other flying insects.

Oviposition decisions by herbivorous insects hinge on multiple factors, with some of the most important being enemy-free space and competition for resources. It is important to understand whether and how herbivores and predators can influence the maternal egg-laying preference when they are alone and in combination with host plants. Here, we evaluate whether the presence of aphids (a competitor) or a lady beetle larvae (a predator) influence host plant selection by an ovipositing butterfly. Canola (Brassica napus L.) was the highest quality of three putative Brassicaceae host plants for aphids Myzus persicae (Sulzer) (Hemiptera: Aphididae), while the butterfly Pieris rapae (L.) (Lepidoptera: Pieridae) showed similar survival on all. Canola was used to determine that the presence of a competitor herbivore (aphids) had no effect on butterfly oviposition behavior. However, predators significantly influenced the number of eggs laid on the plants, especially on those plants that had both aphids and a lady beetle larva present in combination. We expect that adult female P. rapae did not lay their eggs on the treatment that involved both herbivorous competition and predation risk, due to the combined risk factors along with the volatile chemicals and aphid alarm pheromones emitted on those plants that contained both the aphids and lady beetle larva.

Treatment of plants with exogenous salicylic acid (SA) improves resistance to many bacterial pathogens, but can suppress resistance to insect herbivores. While plants vary naturally in constitutive SA, whether such differences are predictive of resistance to insect herbivores has not been studied previously. We examined the possible role of this endogenous SA in structuring the interactions between the cabbage white butterfly, Pieris rapae, and ten hosts in the mustard family (Brassicaceae). Because P. rapae has multiple generations that utilize different hosts across the year, we included five spring-flowering mustards and five summer-flowering mustards that co-occur in ruderal habitats in upstate New York. Under common garden conditions, the spring flowering mustards (Capsella bursa-pastoris, Draba verna, Cardamine impatiens, Barbarea vulgaris, and Arabidopsis thaliana) were significantly more resistant to P. rapae, supporting 42 % less herbivory (P = 0.015) and 64 % lower relative growth rates (P = 0.007), relative to the summer flowering mustards (Sisymbrium altissimum, Brassica nigra, Sinapis arvense, Lepidium campestre, and Arabis canadensis). Leaf total constitutive SA explained significant variation in larval herbivory (R2 = 75.3 %, P = 0.007) and relative growth rates (R2 = 59.4 %, P = 0.043). The three species with the lowest levels of constitutive SA (Capsella bursa-pastoris, Draba verna, and Cardamine impatiens) were the most resistant to larvae. Barbarea vulgaris and Arabis canadensis were notable exceptions, exhibiting high SA concentrations and intermediate resistance to P. rapae. These results suggest a curvilinear relationship between leaf constitutive SA and the herbivory by P. rapae, and they provide some insight into the ecology and possible management of this economically important crop pest. Electronic supplementary material The online version of this article (doi:10.1007/s10886-015-0572-z) contains supplementary material, which is available to authorized users.

Glucosinolates may deter generalist insect feeding as their toxicity causes fitness damage, whereas insects specialized in brassicaceous plants may circumvent the toxic effect. By using no-choice leaf tests, we investigated whether larval development time, body weight, mortality and feeding rate of the generalist Mamestra brassicae (Lepidoptera, Noctuidae) and the specialist Pieris rapae (Lepidoptera, Pieridae), were affected by six genotypes of Brassica oleracea var. acephala, selected for having high or low concentration of sinigrin, glucoiberin (aliphatics) and glucobrassicin (indole). Two phenological plant stages were used. On young plants, M. brassicae most consumed the high sinigrin and low glucoiberin genotypes. Larvae weighed more on the high sinigrin plants. Development time took longer on the low glucoiberin genotype. On mature plants, consumption rate decreased on the high glucoiberin genotype. Larval weight decreased on the high sinigrin, glucoiberin and glucobrassicin genotypes, and development time increased with high glucobrassicin concentration. Pupal weight and mortality rate increased on mature plants, irrespective of the genotype. Pieris rapae fed most on young plants with high sinigrin, and larval weight increased on the high glucoiberin genotype. Mortality increased with low glucoiberin and low glucobrassicin. On mature plants, larval weight decreased with high sinigrin and glucoiberin. The high glucoiberin genotype was the less consumed and also induced a longer development time. High content of aliphatic glucosinolates offered mature plants significant antibiosis defence against both the lepidopterans, whereas the indole glucosinolate was marginally effective. Young plants were more consumed and increased larval weight likely because glucosinolate concentration was still not optimal.

The preference-performance hypothesis posits that the host plant range of plant-feeding insects is ultimately limited by larval costs associated with feeding on multiple resources, and that female egg-laying preferences evolve in response to these costs. The trade-off of either using few host plant species and being a strong competitor on them due to effective utilization or using a wide host plant range but being a poor competitor is further predicted to result in host plant specialization. This follows under the hypothesis that both females and offspring are ultimately favoured by utilizing only the most suitable host(s). We develop an experimental approach to identify such trade-offs, i.e. larval costs associated with being a host generalist, and apply a suite of experiments to two sympatric and syntopic populations of the closely related butterflies Pieris napi and Pieris rapae. These butterflies show variation in their level of host specialization, which allowed comparisons between more and less specialized species and between families within species. Our results show that, first, the link between female host preference and offspring performance was not significantly stronger in the specialist compared to the generalist species. Second, the offspring of the host plant specialist did not outperform the offspring of the generalist on the former's most preferred host plant species. Finally, the more generalized species, or families within species, did not show higher survival or consistently higher growth rates than the specialists on the less preferred plants. Thus, the preference and performance traits appear to evolve as largely separated units.

Population sizes and range of the native butterfly Pieris oleracea declined after habitat loss and parasitism by an exotic braconid wasp (Cotesia glomerata) introduced to control the exotic invasive butterfly Pieris rapae. Further declines are attributed to the invasive exotic weed garlic mustard (Alliaria petiolata), an oviposition sensory trap on which P. oleracea larval survival and growth are very poor. But a population of P. oleracea has adapted to garlic mustard over the past several decades, coincident with the introduction of a second parasitoid, C. rubecula, a specialist on P. rapae that is competitively dominant to C. glomerata. We used stochastic simulation models to assess the plausibility of a hypothesis that reduced parasitoid pressure over this time period enabled P. oleracea to adapt to A. petiolata. We simulated scenarios of trait proliferation via spontaneous mutation or immigration of the trait, and residual variation in the trait following the butterfly’s isolation in North America. Results indicate that the most likely scenario for the population that has adapted to garlic mustard includes (1) a change in selection following garlic mustard invasion to favor previously neutral residual variation in the population, (2) release from parasitism, and (3) evolution of improved larval survival on garlic mustard, which allowed an increased host range, and potentially, population size.

We manipulated the density of the wolf spider, Pardosa astrigera L. Koch in enclosed cabbage plots (2 × 2 m) to assess the impact on the herbivore density and crop yield. P. astrigera was introduced to half of the enclosures at densities of 25 and 50 individuals/m2 in the spring and autumn experiments, respectively; spiders were removed from other enclosures. The results indicated that P. astrigera significantly suppressed the densities of the diamondback moth, Plutella xylostella (L.), in the spring, and the green peach aphid, Myzus persicae (Sulzer), in the autumn experiments. The cabbage yield did not increase, despite the suppression of these herbivores. P. astrigera had no impact on three other lepidopteran species: Pieris rapae crucivora Boisduval, Autographa nigrisigna (Walker), and Mamestra brassicae (L.). The number of spiders increased by 23 % but decreased by 62 % at the end of the spring and autumn experiments, respectively, compared with the initial numbers introduced. Our results show that wolf spiders (Lycosidae) would be effective biocontrol agents of certain herbivore species if spiders abundantly occur early in the crop season and remain stable throughout the season. However, further investigations are required to determine the effect of spiders on the crop yield.

While the phenomenon of male mate choice has attracted considerable attention in the last two decades, whether this sexual selection mechanism could drive the evolution of female ornaments remains poorly understood. Here, we used experimental manipulation of female wing coloration to investigate male mate choice in Pieris rapae, a gift-giving butterfly. Further, we tested whether males’ nutritional status influenced their mating preferences by subjecting larvae to short periods of starvation. We found that males showed significantly more mating approaches toward control females with more colorful wings (higher pteridine content), and that this preference was strongest in low-nutrition males. Additionally, a study of field-collected females revealed that pteridine-based wing coloration was positively correlated with female egg load, which suggests such ornaments may signal female quality. Pteridine-based ornaments are widespread in nature, however their potential as honest signals in male mate choice remains largely unexplored. This work furthers our understanding of how male mate choice and female ornamentation may evolve in species whose mating systems include nutritional nuptial gifts.

The native pierid butterfly Pieris oleracea underwent a large range reduction in New England in the twentieth century, likely due to the introduction the invasive butterfly Pieris rapae (Lep.: Pieridae) to North America in 1860, and later the oligophagous parasitoid Cotesia glomerata (Hymenoptera: Braconidae) in 1884. Thought extirpated from the state by the 1970s, one large dense population of the butterfly was found in the mid 1980s in a flood plain meadow along the Housatonic River in Lenox, Berkshire Co., Massachusetts. We examined how this native pierid was able to maintain a relatively dense local population by feeding on a novel, invasive host plant, Cardamine pratensis (cuckoo flower), in a meadow habitat despite known parasitoid presence. We approached this question in three ways. First, we deployed trap host plants (cuckoo flower and collards) stocked host larvae (first and second instars of either P. rapae or P. oleracea) at the Lenox site and other locations to determine current rates of C. glomerata attack, for comparison with historical information. Second, we used olfactometer experiments to determine if C. glomerata females could detect the cuckoo flower volatiles released during P. oleracea larval feeding. Third, we used field-cage experiments to determine if the plant architecture found in the flood plain meadow inhibited the ability of C. glomerata females to locate and parasitize hosts. Specifically, we asked if overtopping vegetation prevented or reduced parasitism of P. oleracea larvae feeding on the covered basal rosettes of C. pratensis, which is the physical form of host plant for three of the four butterfly generations at the site.

Non-lethal methods of tissue sampling are increasingly used for genetic studies of insect species and the effects of this approach have long been assumed to be minimal. Tissue removal has the potential to influence insect reproductive behaviours such as mate recognition, courtship or oviposition but the effects of non-lethal sampling on reproductive success have not been widely and adequately tested. Here, we test potential effects of both wing-clipping and leg removal on reproductive behaviours of the cabbage white butterfly (Pieris rapae). We conducted a total of 93 male and 59 female mating trials, and found no significant differences in mating success between treated (i.e., tissue removed) and control individuals in either sex. We also monitored the number and location of eggs laid by 58 females. We found no significant differences in egg-laying behaviour among leg removed and control individuals. Power analysis indicated that we had sufficient statistical power to detect moderate effects of treatment on both mating and oviposition. Our study provides the most comprehensive examination to date of the effects of non-lethal sampling on reproductive behaviours in a butterfly/insect species, and supports the contention that tissue sampling is non-detrimental. To fully comprehend the general impacts of tissue sampling on butterfly reproductive behaviour however, additional similar studies need to be conducted on a variety of species with differing mating behaviours. Only through meta-analysis, may it be possible to detect more subtle effects of tissue removal which cannot be revealed within a single study due to sample size limitations.

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.

A general prediction of the specialist/generalist paradigm indicates that plant responses to insect herbivores may depend on the degree of ecological specialization of the insect attacker. However, results from a single greenhouse experiment evaluating the responses of the model plant Arabidopsis thaliana to three specialist (Plutella xylostella, Pieris rapae, and Brevicoryne brassicae) and three generalist (Trichoplusia ni, Spodoptera exigua, and Myzus persicae) insect species did not support the previous prediction. Using an ecological genomic approach, we assessed plant responses in terms of herbivore-induced changes in genome-wide gene expression, defense-related pathways, and concentrations of glucosinolates (i.e., secondary metabolites that are ubiquitously present in cruciferous plants). Our results showed that plant responses were not influenced by the degree of specialization of insect herbivores. In contrast, responses were more strongly shaped by insect taxa (i.e., aphid vs. lepidopteran species), likely due to their different feeding modes. Interestingly, similar patterns of plant responses were induced by the same insect herbivore species in terms of defense signaling (jasmonic acid pathway), aliphatic glucosinolate metabolism (at both the gene expression and phenotypic levels) and genome-wide responses. Furthermore, plant responses to insect herbivores belonging to the same taxon (i.e., four lepidopteran species) were not explained by herbivore specialization or phylogenetic history. Overall, this study suggests that different feeding modes of insect taxa as well as herbivore-specific plant responses, which may result from distinct ecological/evolutionary interactions between A. thaliana (or a close relative) and each of the lepidopteran species, may explain why observed responses deviate from those predicted by the specialist/generalist paradigm.

Food plant preferences of some Lepidoptera species associated with particular colour of the flowers were investigated. Based on 1,329 field observations of 43 Lepidoptera and 66 plant species, Lepidoptera showed a high tendency (G-test, Gadj = 698.6, df = 6, P < 0.001) to use the yellow (29%) and pink (28%) coloured flowers for foraging. Compared to the other colours it was evident that plants with red flowers (2%) were not preferred. Moreover, the plants with red (H = 0.435) and yellow-white (H = 0.543) flowers were not visited by diverse Lepidoptera species. Although yellow and pink flowers were most frequently visited, the highest degrees of the Lepidoptera diversity values were associated with the plants having blue (H = 0.647) and purple (H = 0.634) flowers. Species of Nymphalidae were most numerous (14 spp.) in the study area and the members of this family were observed 430 times on 39 different plant species, but never on plants with red flowers. Pieris rapae was the most abundant species that occurred 136 times on a total of 21 different plant species of which eight had yellow flowers. But, this species has never been seen while feeding on red flowers.

The indirect defense mechanisms of plants comprise the production of herbivore-induced plant volatiles that can attract natural enemies of plant attackers. One of the often emitted compounds after herbivory is methyl salicylate (MeSA). Here, we studied the importance of this caterpillar-induced compound in the attraction of the parasitoid wasp Diadegma semiclausum by using a mutant Arabidopsis line. Pieris rapae infested AtBSMT1-KO mutant Arabidopsis plants, compromised in the biosynthesis of MeSA, were more attractive to parasitoids than infested wild-type plants. This suggests that the presence of MeSA has negative effects on parasitoid host-finding behavior when exposed to wild-type production of herbivore-induced Arabidopsis volatiles. Furthermore, in line with this, we recorded a positive correlation between MeSA dose and repellence of D. semiclausum when supplementing the headspace of caterpillar-infested AtBSMT1-KO plants with synthetic MeSA. Electronic supplementary material The online version of this article (doi:10.1007/s10886-010-9787-1) contains supplementary material, which is available to authorized users.

The evolution of perfect adaptive phenotypic plasticity of a given trait may be influenced by, among other things, phenotypic costs associated with the expression of a given trait value, relative to alternative trait values. One potential cause of such phenotypic costs is the allocation of limited resources to multiple traits. When multiple traits rely on the same resource, trait values for one adaptively plastic trait might be unavoidably associated with maladaptive trait values for other traits. I address this problem in three traits of Pieris rapae L. (the small cabbage white butterfly) that all rely on the pigment melanin and are adaptively plastic, but have very different functions: wing pattern, immune defense, and pupal color. Cool, short-day rearing conditions simultaneously increased total wing melanization and decreased a melanin-based immune response in females, consistent with predictions. However, cool, short days also reduced the melanin-based immune response in males, despite little effect on male wing melanization. Furthermore, contrary to predictions, these patterns were not altered by differences in dietary resources. Finally, dark-colored rearing backgrounds during pupation substantially increased pupal melanization in both sexes, but was not associated with differences in wing melanization. These results offer only mixed support for the hypothesis of melanin-based trade offs as a source of phenotypic costs to adaptive plasticity in these traits. However, patterns of sexual dimorphism for these traits suggest trade offs might be at work at another level: relative to males, females have consistently more heavily melanized wings but less heavily melanized pupae and immune responses. The reduced immune response under cool, short-day conditions may also have implications for the evolutionary ecology of these butterflies.

Encroachment by invasive plants is a leading threat to rare butterflies. Restoration plans increasingly recommend herbicides to control invasive plants within butterfly habitats. Few studies address the effects of these herbicides on at-risk butterflies. The effects of two graminicides (fluazifop-p-butyl and sethoxydim) and a surfactant (Preference®) were evaluated on Icaricia icarioides blackmorei and Pieris rapae. The effects on butterfly larvae were assessed by mimicking recommended timing and mixture rates of field applications. Differences in survival to adult eclosure, development time, biomass, sex ratio and adult morphology were assessed. Survival of P. rapae was reduced by 32% with sethoxydim and 21% with fluazifop-p-butyl. Wing size and pupal weights of P. rapae were reduced by herbicide treatments. Icaricia icarioides blackmorei experienced a 21% reduction in development time from the date of treatment to eclosure. These results highlight the importance of careful consideration in the use of herbicides in habitats harboring at-risk butterfly populations.

We analyzed a portion of mitochondrial COI gene sequences (658 bp) to investigate the genetic diversity and geographic variation of the swallowtail butterfly, Papilio xuthus L. (Lepidoptera: Papilionidae), and the cabbage butterfly, Pieris rapae L. (Lepidoptera: Pieridae). Papilio xuthus showed a moderate level of sequence divergence (0.91% at maximum) in 15 haplotypes, whereas Pi. rapae showed a moderate to high level of sequence divergence (1.67% at maximum) in 30 haplotypes, compared with other relevant studies. Analyses of population genetic structure showed that most populations are not genetically differentiated in both species. The distribution pattern of both species appears to be consistent with category IV of the phylogeographic pattern sensu Avise: a phylogenetic continuity, an absence of regional isolation of mtDNA clones, and extensive distribution of close clones. The observed pattern of genetic diversity and geographic variation of the two butterfly species seem to reflect the abundant habitats, abundant host plants, and flying abilities in connection with the lack of historical biogeographic barriers.

Pieris rapae extract was sprayed on the surface of cucumber leaf to determine the induction of resistance to anthracnose. The enzyme activities of peroxidase (POD) and polyphenoloxidase (PPO) were detected on cucumber leaves after P. rapae extract induction and pathogen challenge. The results showed that the disease index of cucumber anthracnose was significantly decreased after the cucumber was induced with the P. rapae extract at a concentration of 5.0 mg·mL−1. The POD and PPO activities in foliar-applied P. rapae extract without pathogen inoculation (PETO) or with pathogen inoculation (PETI) were relatively higher than those with no-P. rapae extract treatment and without pathogen inoculation (CONO) or with pathogen inoculation (CONI), respectively. The results suggested that the increased levels of POD and PPO activities in PETO and PETI play an important role in the induction of resistance to cucumber anthracnose.

Many parasitoids are known to use herbivore-induced plant volatiles as cues to locate hosts. However, data are lacking on how much of an advantage a parasitoid can gain from following these plant cues and which factors can limit the value of these cues to the parasitoid. In this study, we simulate the Cotesia rubecula–Pieris rapae–Brassica oleracea system, and ask how many more hosts can a parasitoid attack in a single day of foraging by following plant signals versus randomly foraging. We vary herbivore density, plant response time, parasitoid flight distance, and available host stages to see under which conditions parasitoids benefit from herbivore-induced plant cues. In most of the parameter combinations studied, parasitoids that responded to cues attacked more hosts than those that foraged randomly. Parasitoids following plant cues attacked up to ten times more hosts when they were able to successfully attack herbivores older than first instar; however, if parasitoids were limited to first instar hosts, those following plant cues were at a disadvantage when plants took longer than a day to respond to herbivory. At low herbivore densities, only parasitoids with a larger foraging radius could take advantage of plant cues. Although preference for herbivore-induced volatiles was not always beneficial for a parasitoid, under the most likely natural conditions, the model predicts that C. rubecula gains fitness from following plant cues.

Parasitoids respond to volatiles that plants produce when injured by herbivores. A considerable body of literature addresses the chemical pathways of herbivore-induced volatile production. However, there is almost no theory or data on how timing of volatile release in relationship to host availability for parasitization impacts the utility of these cues to parasitoids and on the extent that this volatile release timing might increase or decrease the percent of herbivores that become parasitized. This kind of information is critical in judging the benefits that might accrue from a breeding program aimed at enhancing herbivore-responsive volatile production. We developed a general model to begin examining this issue by using available parameters from two tritrophic systems. The model uses herbivore oviposition, development, and mortality rates, linked to a range of plant volatile induction and cessation periods for calculating the proportion of plants in a field that are (1) not producing volatiles but occupied by suitable herbivore hosts, (2) producing volatiles and occupied by suitable herbivore hosts, (3) producing volatiles but not occupied by suitable herbivore hosts, and (4) not producing volatiles and not occupied by suitable herbivore hosts. The impact of the plant volatiles on parasitoid foraging success is then determined by comparing the expected number of hosts parasitized when the parasitoid focuses solely on the volatile-producing plants to when it forages randomly among all plants. Under some conditions, parasitoids can attack three times more herbivores if they focus on volatile-producing plants. However, when we simulate plants that take several days to cease volatile production after pupation or death of the herbivore, parasitization rate does not increase when parasitoids use volatiles as cues. The utility of the volatile cues is consistently greater when a smaller proportion of plants is occupied by herbivores, indicating that their usefulness may be reduced to zero in fields saturated with volatiles.

Phenoloxidase (PO) is a key enzyme in insect development, responsible for catalyzing the hydroxylation of tyrosine into o-diphenols and the oxidation of o-diphenols into o-quinones. In the present study, the kinetic assay in air-saturated solutions and the kinetic behavior of PO from Pieris rapae (Lepidoptera) larvae in the oxidation of L-tyrosine (a monophenol) and L-DOPA (l-3, 4-dihydroxyphenylalanine) (a diphenol) was studied. The inhibitory effects of 3-hydroxy-4-methoxybenzaldehyde thiosemicarbazone (3-H-4-MBT) on the monophenolase and diphenolase activities of PO were also studied. The results show that 3-H-4-MBT can inhibit both the monophenolase and diphenolase activities of PO. The lag period of L-tyrosine oxidation catalyzed by the enzyme was obviously lengthened and the steady-state activities of the enzyme sharply decreased. The inhibitor was found to be noncompetitively reversible with a K I (K I = K IS) of 0.30 micromol/L and an estimated IC50 of 0.14 +/- 0.02 micromol/L for monophenolase and 0.26 +/- 0.04 micromol/L for diphenolase. In the time course of the oxidation of L-DOPA catalyzed by the enzyme in the presence of different concentrations of 3-H-4-MBT, the rate decreased with increasing time until a straight line was approached. The microscopic rate constants for the reaction of 3-H-4-MBT with the enzyme were determined.

Caffeine (1,3,7-trimethylxanthine) is one of the most widely used plant secondary metabolites, primarily as a stimulant and an ingredient in drugs. In nature, caffeine is believed to function in chemical defense, acting as an antiherbivory and allelopathic agent, and therefore it might be employed to protect agriculturally important crop plants. In coffee plants, caffeine is synthesized from the precursor xanthosine in four steps, three N-methylations and removal of ribose. We had previously isolated genes encoding three distinct N-methyltransferases, and we demonstrated production of recombinant enzymes that yielded caffeine in in vitro reconstitution experiments. When these caffeine biosynthetic pathway genes were simultaneously expressed in tobacco plants (Nicotiana tabacum), caffeine was successfully produced up to 5 microg/g fresh weight in leaves. The leaves were unpalatable to tobacco cutworms (Spodoptera litura). This repellent action appeared to be more widely applicable to lepidopteran caterpillars as observed with small white (Pieris rapae) fed on Chinese cabbages that had been top-treated with caffeine. Our recent results suggest a novel approach to strengthen anti-herbivore traits by producing caffeine in crop plants.

The thale cress, Arabidopsis thaliana, is considered to be an important model species in studying a suite of evolutionary processes. However, the species has been criticized on the basis of its comparatively small size at maturity (and consequent limitations in the amount of available biomass for herbivores) and on the duration and timing of its life cycle in nature. In the laboratory, we studied interactions between A. thaliana and the cabbage butterfly, Pieris rapae, in order to determine if plants are able to support the complete development of the herbivore. Plants were grown in pots from seedlings in densities of one, two, or four per pot. In each treatment, one, two, or five newly hatched larvae of P. rapae were placed on fully developed rosettes of A. thaliana. In a separate experiment, the same densities of P. rapae larvae were reared from hatching on single mature cabbage (Brassica oleracea) plants. Pupal fresh mass and survival of P. rapae declined with larval density when reared on A. thaliana but not on B. oleracea. However, irrespective of larval density and plant number, some P. rapae were always able to complete development on A. thaliana plants. A comparison of the dry mass of plants in different treatments with controls (= no larvae) revealed that A. thaliana partially compensated for plant damage when larval densities of P. rapae were low. By contrast, single cress plants with 5 larvae generally suffered extensive damage, whereas damage to B. oleracea plants was negligible. Rosettes of plants that were monitored in spring, when A. thaliana naturally grows, were not attacked by any insect herbivores, but there was often extensive damage from pulmonates (slugs and snails). Heavily damaged plants flowered less successfully than lightly damaged plants. Small numbers of generalist plant-parasitic nematodes were also recovered in roots and root soil. By contrast, plants monitored in a sewn summer plot were heavily attacked by insect herbivores, primarily flea beetles (Phyllotreta spp.). These results reveal that, in natural populations of A. thaliana, there is a strong phenological mismatch between the plant and most of its potential specialist insect herbivores (and their natural enemies). However, as the plant is clearly susceptible to attack from non-insect generalist invertebrate herbivores early in the season, these may be much more suitable for studies on direct defense strategies in A. thaliana.

To elucidate the role of the plant lipoxygenase (LOX)/lyase pathway for host search behavior of two parasitic wasps attacking herbivorous larvae, an Arabidopsis mutant (all84) was isolated with a mutation somewhere in the LOX/lyase pathway. Detached leaves of the mutant were shown to release less (Z)-3-hexenal, a first green leaf volatile (GLV) product of the LOX/lyase pathway. The braconid larval parasitoids studied, Cotesia glomerata and Cotesia plutella, differ in their ability to discriminate among plant volatiles induced by feeding of lepidopteran hosts and nonhosts: C. plutella only responds to plant volatiles induced by hosts (Plutella larvae), whereas the response by the more generalist C. glomerata is not host specific. The Arabidopsis mutant all84 infested by Pieris larvae was less attractive to C. glomerata than Arabidopsis wild type (wt) infested by the host larvae. C. glomerata was attracted by two of the GLV biosynthesized through the LOX/lyase pathway, (E)-2-hexenal and (Z)-3-hexenyl acetate. However, attraction of C. plutellae to volatiles from Plutella-infested all84 plants did not differ from attraction to host-infested wt Arabidopsis. Both wasp species were arrested to the respective host-infested edge of the wt leaf by showing characteristic antennal searching behavior on the edge. In C. glomerata, the duration of this searching behavior at the infested leaf edge was significantly shorter on all84 plants than on wt plants. By contrast, the duration of the searching behavior of C. plutellae on the host-infested leaf edge of all84 was not significantly different from that on the wt leaf. These data suggest that the LOX/lyase pathway is directly involved in the production of attractants and arrestants important for host search behavior of the more generalist C. glomerata, but not for the specialist C. plutellae.

Ascoviruses (AVs) infect larvae of various insect pests belonging to the family Noctuidae. The result of AV infection in the hosts is cleavage of infected cells into vesicles, a unique feature of AV infection. Since insect cell lines facilitate the study of virus life cycles, attempts were made to analyze Heliothis virescens AV (HvAV3e) infection in several cell lines and compare cell pathology to larval infection. In this study, replication and cytopathological effects of HvAV3e on four different cell lines were investigated. HvAV3e replication was confirmed in three noctuid cell lines from Spodoptera frugiperda (Sf9) and Helicoverpa zea (BCIRL-Hz-AM1 and FB33). However, the virus did not replicate in the non-noctuid insect cell line from Pieris rapae (Pieridae). Despite replication of the virus in the three permissive cell lines, the cytopathological effects of the virus were significantly different from that of larval infection.

Larvae of the cabbage white Pieris rapae are specialists on plants belonging to the family Brassicaceae (Cruciferae). Adult females have been shown to use the glucosinolate gluconasturtiin (phenylethylglucosinolate) as a recognition cue for cruciferous plants, so they can identify an appropriate host for oviposition (Huang and Renwick in J Chem Ecol 20:1025-1037, 1994). Here, we report our results from a study of the role of this glucosinolate in feeding preferences of P. rapae larvae. The larvae were allowed to choose between leaf disks from the non-host cowpea Vigna sinensis (Fabaceae) that were treated with pure gluconasturtiin in solvent, or solvent alone. Our results showed that gluconasturtiin is a feeding stimulant for P. rapae larvae. A series of chemosensory ablations revealed that this response is mediated by one set of taste sensilla, the sensilla styloconica. Electrophysiological tip recordings revealed two neurons in the lateral sensillum styloconicum that are sensitive to gluconasturtiin. These neurons show significantly higher firing frequencies with 4 mM gluconasturtiin added to the recording pipette than for recording solution alone. We propose that the sensitivity to gluconasturtiin shown by these two taste neurons is an important contributor to the animals' behavioral preference for this compound.

Effect of GCSC-BtA (Germany-China Scientific Cooperation — Bacillus thuringiensis — Abamectin) biocide on abundance and diversity of some important cabbage pests, i. e., Plutella xylostella (L.) (Lep., Plutellidae), Pieris rapae (L.), (Lep., Pieridae), Brevicoryne brassicae (L.) (Hom., Aphididae), Liriomyza sativae Blanch. (Dip., Agromyzidae), Phyllotreta vittata Fabric. (Col., Chrysomelidae) as well as their natural enemies, i. e., Apanteles plutellae Kurdj. (Hym., Braconidae), Erigonidium graminicola (Sundv.) (Araneida, Linyphiidae), Coccinella septempunctata L. (Col., Coccinellidae), were determined in comparison to that of methomyl insecticide in cabbage fields in Fuzhou region of the southeastern China.

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.

We experimentally reanalyzed the classic interaction between Pieris rapae, a specialist lepidopteran herbivore, and isothiocyanates (mustard oils) that are characteristic phytochemicals of the Brassicaceae. Previous investigations have suggested that P. rapae is unaffected by isothiocyanates. Using whole plants, root extracts, and a microencapsulated formulation of allyl isothiocyanate, we now show that isothiocyanates reduce herbivore survival and growth, and increase development time, each in a dose-dependent manner. Neither the substrate allyl glucosinolate, nor myrosinase, the enzyme that results in the breakdown of glucosinolates, negatively affected P. rapae. Thus, we present strong evidence for a role for isothiocyanates in plant resistance against the specialist herbivore P. rapae.

Male Pieris napi butterflies previously have been shown to synthesize and transfer an antiaphrodisiac, methyl salicylate (MeS), to females at mating. This substance curtails courtship and decreases the likelihood of female remating. Here, we show that similar systems occur in Pieris rapae and Pieris brassicae. In P. rapae, 13C-labeling studies showed that males utilize the amino acids phenylalanine and tryptophan as precursors to MeS and indole, respectively. These volatiles are transferred to females at mating and function as antiaphrodisiacs, as demonstrated by field tests entailing painting MeS, indole, or a mixture on the abdomens of virgin females and assessing their attractiveness to wild males. With P. brassicae, 13C-labeling studies showed that males use phenylalanine as a precursor to synthesize benzyl cyanide, which was demonstrated to function as an antiaphrodisiac by field tests similar to those for P. rapae. This communication system exhibits both similarities and differences among the three species; in P. napi and P. rapae, males are fragrant but transfer a volatile antiaphrodisiac to females that is completely different from the male odor, whereas in P. brassicae the antiaphrodisiac transferred by male to female is identical with male odor.

Specificity of plant induction responses may be important to the interactions between mustards and insect herbivores. This study compared the effects of the cabbage white butterfly, Pieris rapae (L.), cabbage looper, Trichoplusia ni (Hubner), and the mustard flea beetle, Phyllotreta cruciferae (Goeze) on induction of leaf trichome density, sinigrin concentration, and nitrogen concentration in black mustard, Brassica nigra (L.) Koch. Plants were damaged for 12 h at the four-leaf stage, with effort made to standardize the damage applied. Induction responses were measured on the fifth, seventh, ninth, and 11th leaves counted from the cotyledons. Seventh leaves of plants damaged by P. rapae had 76% more trichomes per unit area than controls, whereas equivalent leaves of plants damaged by the other two herbivores exhibited no response. Ninth leaves of plants damaged by T. ni had 113% more trichomes per unit area than controls, whereas equivalent leaves of plants damaged by the other two herbivores exhibited no response. Trichome densities of fifth and 11th leaves did not respond to treatments. Leaf sinigrin and nitrogen concentrations were not affected by the damage treatments. Differential plant trichome response to P. rapae and T. ni may have been due to differences in location of feeding during the damage treatment. Other cues, such as salivary components, may also have differed between the two herbivores. This study is one of the first to document differential effects of two herbivores from the same guild on induction of morphological resistance.

The epicuticular composition of different body parts of the Cabbage White, Pieris rapae L., was investigated using GC and GC/MS. The major group of components, hydrocarbons, occurs in two distinct classes, which show different distributions on the cuticle of the insects. Unbranched shorter chain compounds (C21 to C31, linear group) dominate on body, head and wings, while longer chain, polymethyl-branched compounds (C35 to C39, branched group) are predominantly found on the antennae. Several other components like 1,3-pentacosadiene and oxygenated aliphatic compounds occur in minor amounts on the cuticle. The reason for this polymorphism is discussed.

A synthetic Bacillus thuringiensis cry1C gene was transferred to three Korean cultivars of Chinese cabbage via Agrobacterium tumefaciens-mediated transformation of hypocotyl explants. Hygromycin resistance served as an efficient selective marker. The transformation efficiency ranged from 5% to 9%. Transformation was confirmed by Southern blot analysis, PCR, Northern analysis, and progeny tests. Many transgenic plants of the closed-head types (lines Olympic and Samjin) flowered in vitro. Over 50 hygromycin-resistant plants were successfully transferred to soil. The transgenic plants and their progeny were resistant to diamondback moths (DBM, Plutella xylostella), the major insect pest of crucifers world-wide, as well as to cabbage loopers (Trichoplusia ni) and imported cabbage worms (Pieris rapae). Both susceptible Geneva DBM and a DBM population resistant to Cry1A protein were controlled by the Cry1C-transgenic plants. The efficient and reproducible transformation system described may be useful for the transfer of other agriculturally important genes into Chinese cabbage.

The host ranges of phytophagous insects are determined to a large degree by plant chemistry. Specialist insects are often closely associated with plants that produce characteristic chemicals, which may act as attractants or stimulants to aid in finding or recognizing a host. Generalist insects are generally believed to rely on the presence of repellents or deterrents to ensure avoidance of unsuitable plants. However, the chemistry of any plant can be highly variable, as a result of growth characteristics, genetic variation, or environmental factors. Such variable chemistry may provide windows of opportunity for nonadapted insects to utilize a plant or for a plant to become resistant to a normally adapted herbivore. Differences in insect responses to plant constituents may also result from genetic variation or environmental factors. In particular, dietary experience has been found to influence the ability of insects to taste plant chemicals that may serve as signals of suitability or unsuitability. Certain dietary constituents appear to suppress the development of taste sensitivity to deterrents in an insect, whereas the presence of specific stimulants in the diet may result in the development of dependence on these compounds. These findings further emphasize the fact that the dynamics of plant biochemistry along with plasticity in the sensory system of insects might be expected to play a major role in the evolution of new plant–insect relationships.

Few studies have investigated insect egg-laying preferences in relation to photosynthesis or transpiration of their host plants. It has been suggested that intravarietal preferences of the small white butterfly (Pieris rapae L.: Pieridae) include larger plants with characteristically higher transpiration rates. Interestingly this species, like many other Lepidoptera, may detect biogenic CO2 gradients associated with photosynthesis. We studied egg-laying preferences in working farm environments examining relationships among host choice, plant gas exchange activity, and plant size. Females discriminated between plants in monocultures on the basis of height. A balance of pre- and post–alighting preferences resulted in plants of medium size receiving eggs. Post–alighting preferences led to plants, but not alighted leaves, with higher rates of photosynthesis supporting eggs. These findings do not support a mechanistic basis for the use of gas exchange activity during host selection but, for the first time, indicate the greater physiological activity of crop plants that ultimately received the eggs of a pest insect.

Many plant species defend themselves against herbivorous insects indirectly by producing volatiles in response to herbivory. These volatiles attract carnivorous enemies of the herbivores. Research on the model plant Arabidopsis thaliana (L.) Heynh. has contributed considerably to the unraveling of signal transduction pathways involved in direct plant defense mechanisms against pathogens. Here, we demonstrate that Arabidopsis is also a good candidate for studying signal transduction pathways involved in indirect defense mechanisms by showing that: (1) Adult females of Cotesia rubecula, a specialist parasitic wasp of Pieris rapae caterpillars, are attracted to P. rapae-infested Arabidopsis plants. (2) Arabidopsis infested by P. rapae emits volatiles from several major biosynthetic pathways, including terpenoids and green leaf volatiles. The blends from herbivore-infested and artificially damaged plants are similar. However, differences can be found with respect to a few components of the blend, such as two nitriles and the monoterpene myrcene, that were produced exclusively by caterpillar-infested plants, and methyl salicylate, that was produced in larger amounts by caterpillar-infested plants. (3) Genes from major biosynthetic pathways involved in volatile production are induced by caterpillar feeding. These include AtTPS10, encoding a terpene synthase involved in myrcene production, AtPAL1, encoding phenylalanine ammonia-lyase involved in methyl salicylate production, and AtLOX2 and AtHPL, encoding lipoxygenase and hydroperoxide lyase, respectively, both involved in the production of green leaf volatiles. AtAOS, encoding allene oxide synthase, involved in the production of jasmonic acid, also was induced by herbivory.

Field observations revealed that flowers of Osmanthus fragrans attract few insect species despite their strong scent and vivid coloration (yellowish orange). Floral volatiles of the plant were thus examined for suspected repellency to Pieris rapae, a potential visitor that never visits this flower. The influence of odor components on the foraging behavior of adult butterflies was assessed by two behavioral bioassays: proboscis extension reflex (PER) in response to olfactory stimuli and the frequency of visits to artificial flowers. In the PER bioassays, a binary mixture composed of a test material and 2-phenylethanol (a positive standard compound) was used, and the negative effect of test materials was appraised based on the degree of suppression caused compared with the original PER performance evoked by 2-phenylethanol alone. Isopentane extract of the flower exhibited significant deterrency in the PER test and remarkable repellency in the flower-visiting experiment, thereby suggesting the presence of a repellent(s) in the floral volatiles. Fractionation of the extract by silica gel column chromatography yielded an active fraction (based on PER performance), which, upon GC and GC-MS analyses, was found to consist of γ-decalactone, β-ionone, and linalool oxide (LO) isomers (furanosides and pyranosides) as major components together with small amounts of linalool and α-ionone. γ-Decalactone was strongly deterrent in the PER test, and β-ionone and all LO isomers were weakly deterrent, while the other compounds exhibited no deterrency. In the flower-visiting tests, γ-decalactone showed notable repellency, whereas β-ionone had no repellent effect. Among the LO isomers, at least furanosides (cis/trans mixture) and cis-pyranoside were found to be weakly repellent. On the other hand, the butterfly showed strong to weak EAG responses to LO furanosides, cis-LO pyranoside, γ-decalactone, and β-ionone in decreasing order of intensity. The present results clearly indicate that the repellency of a compound is correlated with its deterrent effect on PER but not necessarily with antennal sensitivity.

A synthetic Bacillus thuringiensis (Bt) cry1C gene was introduced into broccoli (Brassica oleracea ssp. italica) by Agrobacterium-mediated transformation. Twenty-one Cry1C transgenic plants were regenerated from 400 hypocotyl and petiole explants. Variable amounts of stable steady- state cry1C mRNA accumulated in different transgenic plants. Cry1C protein (up to 0.4% of total soluble protein) was produced in correlation with the cry1C mRNA levels. Leaf section and whole-plant bioassays were done using diamondback moth (DBM) larvae from lines susceptible to Bt or resistant to Cry1A or Cry1C proteins (Cry1AR or Cry1CR, respectively). Plants with high levels of Cry1C protein caused rapid and complete mortality of all three types of DBM larvae with no defoliation. Plants with lower levels of Cry1C protein showed an increasing differential between control of susceptible of Cry1AR DBM. This study demonstrated that high production of Cry1C protein can protect transgenic broccoli not only from susceptible or Cry1AR DBM larvae but also from DBM selected for moderate levels of resistance of Cry1C. The Cry1C- transgenic broccoli were also resistant to two other lepidopteran pests of crucifers (cabbage looper and imported cabbage worm). These plants will be useful in studies of resistance management strategies involving multiple transgenes.

Scent and coloration of corolla were examined as floral attributes responsible for preferential visiting by the cabbage butterfly, Pieris rapae, to rape flower, Brassica rapa. Floral volatile components that release the flower-visiting behavior of the butterfly were identified by chemical analyses, electroantennography (EAG), and two behavioral bioassays: proboscis extension reflex (PER) in response to odor and attraction to artificial flowers. GC and GC-MS analyses of the headspace volatiles from the flowers revealed the presence of six aromatic compounds, benzaldehyde, phenylacetaldehyde, benzyl alcohol, 2-phenylethanol, phenylacetonitrile, and indole in decreasing order of quantity. Of these, phenylacetaldehyde elicited the highest response in the PER assay. While benzyl alcohol, 2-phenylethanol, benzaldehyde, and phenylacetonitrile evoked moderate responses, the PER-eliciting activity of indole was very weak. In two-choice behavioral bioassays, artificial flowers scented with any one of these PER-active compounds attracted significantly more butterflies than control (unscented) flowers, whereas those treated with indole were almost inactive. The EAG activities of the six chemicals were not high and were about the same at a low dose (1 μg), but phenylacetaldehyde elicited a much stronger response from both sexes at higher doses (10 and 100 μg). An overall profile of EAG responses at a dose of 100 μg was analogous to that of PER performance, suggesting that floral volatiles may be involved in close-range location or recognition of flowers rather than long-range attraction. By spectroscopic and UV-photographic examinations of rape flower, the central part of the corolla was found to absorb UV rays in marked contrast to the other parts, which reflected near-UV rays (λmax = 350 nm). This indicates that the flower is endowed with a conspicuous nectar guide that is probably an important visual stimulus for attracting foraging adults of P. rapae. Consequently, the present findings strongly suggest that this elaborate pollination strategy of rape flower, characterized by its good combination of olfactory and visual attractiveness, accounts for preferential visiting by the cabbage butterfly to the flower.

Using alternating bands of weeds and broccoli I experimentally manipulated vegetation composition and the spatial scale at which the landscape was fragmented in a factorial design. This experimental approach allowed me to distinguish the effect of spatial scale from that of simple crop heterogeneity on crop herbivores. The importance of scale depended on which insect species were examined. Cabbage aphids (Brevicoryne brassicae) were influenced by vegetation composition at all tested scales of fragmentation; cabbage butterflies (Pieris rapae) were not affected by scale or by composition and flea beetles (Phyllotreta cruciferae) revealed a striking dependence on scale of fragmentation as well as an interaction between scale and composition. This approach shows the importance of dissecting out the effects of scale from other aspects of landscape manipulation, and emphasizes the challenge of developing a theory that will enable prediction of species-specific responses to scale.

Floral scent compounds of Ligustrum japonicum that affect the foraging behavior of Pieris rapae adults were examined by means of chemical analyses, electroantennogram (EAG) responses, and behavioral bioassays; the behavioral biossays consisted of two tests: reflex extension of proboscis (REP) in response to odor, and attraction to scented and unscented artificial flowers. More than 30 compounds, including 2-phenylethanol, benzyl alcohol, and methyl phenylacetate as the major components were identified from L. japonicum flowers. Of these, 22 compounds were tested for their effect on foraging behavior. Phenylacetaldehyde (PA), 2-phenylethanol (PE), and 6-methylhept-5-en-2-one (MHO) elicited the highest REP responses, and benzaldehyde (BA) and methyl phenylacetate (MPA) evoked intermediate REP responses. EAG responses were not necessarily correlated with REP activities; the three high-REP compounds gave only moderate EAG responses, whereas two other compounds (ethyl phenylacetate and 2-phenylethyl acetate) that released high EAG responses showed low REP activities. In two-choice behavioral bioassays, flower models scented with any one of these high-REP compounds attracted significantly more adults, while compounds with low REP activities exhibited weak or no appreciable attractiveness. This suggests that the REP responsiveness closely reflects the attractiveness of a compound and could be an effective measure in elucidating which chemical attractants are involved in flower-visiting. A synthetic blend of five floral chemicals (PA, PE, MHO, BA, and MPA) displayed an attractiveness that was comparable to that of the floral extract and was more effective in attractiveness than the compounds tested singly. Consequently, it is highly likely that the flower-visiting by P. rapae to L. japonicum is mediated largely by floral scent chemicals and that a synergistic effect of the five floral components would be most responsible for attraction of the butterfly to this flower. The present results also strongly suggest that specific floral volatiles may facilitate close-range flower location by P. rapae, could serve in part as a cue for recognizing food sources, and also be profoundly implicated in flower preference.

The effect of repeated matings on sperm numbers in successive ejaculates of the cabbage white butterfly, Pieris rapae, was examined. First ejaculates were larger than successive ones, which did not differ among themselves. Moreover, the cumulative mass of previous spermatophores was not correlated with that of the last mating. The number of eupyrene sperm bundles in the ejaculate did not differ between first and successive matings. Multiplying by 256, a male transfers about 11,000 eupyrene sperm at every mating. First ejaculates contained about 46,000 apyrene sperm, whereas successive ejaculates contained higher numbers. The sperm density increased after the first mating, though the spermatophore mass decreased. The significance of change in sperm quantity with mating number is discussed from the viewpoint of male investment.