Volatiles from damaged plants as major cues in long-range host-searching by the specialist parasitoidCotesia rubecula

The role of volatile stimuli in the long-range host-searching behaviour of the specialist parasitoidCotesia rubecula Marshall (Hymenoptera: Braconidae) was studied. Components from the plant-host-complex Brussels sprouts (Brassica oleracea L. var.gemmifera (DC.) Schulz. cv. ‘Titurel’)-Pieris rapae L. (Lepidoptera: Pieridae) were compared for their attractiveness in dual choice tests in a windtunnel. Stimuli from cabbage plants that were mechanically damaged or damaged byP. rapae caterpillars were more attractive to this parasitoid species than stimuli emitted by the host larvae or their faeces. Parasitoids preferred leaves from the plant-host-complex over artificially damaged leaves. Undamaged cabbage plants were the least attractive to the foraging females. These results indicate that in-flight searching behaviour ofC. rubecula is guided by plant-derived information and that for this specialist species more reliable and specific host-derived cues play a minor role at longer distances.     

Nutritional suitability and ecological relevance of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> and <Emphasis Type="Italic">Brassica oleracea</Emphasis> as foodplants for the cabbage butterfly, <Emphasis Type="Italic">Pieris rapae</Emphasis>

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.     

Host-age discrimination during host location byCotesia glomerata, a larval parasitoid ofPieris brassicae

Some parasitoids are restricted with respect to the host stage that they attack and even to a certain age within a stage. In this paper we investigate whether the parasitoidCotesia glomerata can discriminate between old and young caterpillar instars of its host,Pieris brassicae, before contacting these hosts, since contacts with older instars are very risky with a chance of being killed, due to the aggressive defensive behaviour of the caterpillars. Flight chamber dual choice tests showed that volatile chemicals emitted by Brussels sprouts plants (Brassica oleracea var. gemmifera) after feeding damage by 1st and 5th larval instars are equally attractive to the wasps. Simulated herbivore damage by 2nd and 5th larval instars, obtained by treating mechanically damaged leaves with carterpillar regurgitant, was also equally attractive, even when the wasps were exposed to repeated experience on different larval instars to increase their discriminatory ability. In contrast, single choice contact bioassays showed that the time spent searching on a leaf with feeding damage of 1st instar larvae was significantly longer than the time spent on 5th instar feeding damage or on mechanically damaged leaves. Both flight and contact bioassays did not show any effect of egg-related infochemicals. The results demonstrate thatC. glomerata can discriminate between young and old larval instars ofP. brassicae, without contacting the caterpillars. This is not done through volatile herbivore-induced synomones but through cues that are contacted after arrival at a caterpillar-infested leaf.     

Orientation behaviour of the predatory hemipteran Perillus bioculatus to plant and prey odours

The pentatomid predator P. bioculatus responded by positive odour-conditioned anemotaxis when exposed to airborne volatiles emitted by potato plants damaged by Colorado potato beetle larvae, whereas intact potato plants and non-feeding larvae as odour sources failed to elicit anemotaxis. Walking tracks of adult predators had higher values of straightness and upwind fixation when odours emanating from mechanically damaged plants were encountered than tracks registered in response to air carrying volatiles from intact plants, but these parameters returned to control values within 1–2 h after damage was caused. In contrast, air led over plants damaged by beetle larvae elicited orientation responses at least 3 hours after feeding damage ceased. The combination of chemical data on headspace composition, olfactory sensitivity established in electro-antennogram studies and behavioural data presented here imply a role of sesquiterpenoid plant volatiles as odorous cues eliciting attraction of this predator to damaged potato plants.     

Effects of azadirachtin after systemic uptake intoBrassica oleracea on larvae ofPieris brassicae

Petioles of intact cabbage leaves were placed in azadirachtin solutions for 24 h. Azadirachtin or active metabolites of this compound appeared to be translocated systemically as performance of first instar larvae ofPieris brassicae L. placed on disks from those leaves was affected. The position within a leaf from which disks were excised did not affect the extent of these effects, indicating a homogeneous distribution of active compounds in the laminar tissue. At doses ranging from 1–10 ppm of azadirachtin in the solution taken up by the leaves, gain of larval fresh weight and leaf area consumed were reduced. These effects were dose dependent up to 60–90 ppm. At 10 ppm and higher doses no larva reached the second instar within 72 h. Mortality during the 72 h period was highest at 10 ppm, due to the coincidence of lethal action with ecdysis in combination with the effect on developmental rate. Azadirachtin solutions applied directly to the surface of leaf disks in order to produce known doses on a leaf fresh weight basis gave results similar to the systemic treatment for weight gain, leaf area consumed and developmental rate, while mortality was higher in the latter treatment.     

Volatiles released from bean plants in response to agromyzid flies

Liriomyza sativae Blanchard and Liriomyza huidobrensis (Blanchard) (Diptera: Agromyzidae) are two invasive flies in China that have caused economical damage on vegetables and ornamental plants. In this article, we report the profiles of emitted volatiles from healthy, mechanically damaged, and leafminer-damaged bean, Phaseolus vulgaris L., plants. Among 25 emitted volatiles identified, (E)-2-hexen-1-al, (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), (Z)-3-hexenyl acetate, (Z)-3-hexen-1-ol, (syn)- and (anti)-2-methylpropanal oxime, (syn)-2-methylbutanal oxime, linalool, and (E,E)-α-farnesene were consistently released from damaged bean plants. Combined amounts of these nine compounds made up more than 70% of the total volatiles emitted from each treatment. No qualitative differences in volatile emission were found between bean plants damaged by the two fly species; however, amounts of several major compounds induced by L. huidobrensis damage were significantly higher than those from plants damaged by L. sativae. The mechanically damaged plants released a higher proportion of green leaf volatiles than plants in the other treatments, whereas leafminer-damaged plants produced more terpenoids and oximes. Furthermore, the volatile profiles emitted from plants, damaged by adult leafminers, by second instar larvae, and even the plants with empty mines left by leafminer larvae (the pupal stage) were significantly different. The identification of volatile oximes released from damaged plants was confirmed and is discussed in a behavioral and biological control context.Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.     

Cross habituation to feeding deterrents and acceptance of a marginal host plant byPieris rapae larvae

Sensitivity of the cabbage butterfly,Pieris rapae L. to feeding deterrents was compared for larvae reared on different food sources under laboratory conditions. Since cabbage-reared larvae normally reject nasturtium,Tropaeolum majus L., the effects of previous exposure to allelochemicals on larval acceptance or rejection of this plant were also examined. When compared with cabbage-reared larvae, nasturtium-reared larvae were less sensitive to feeding deterrents including cymarin, erysimoside and 2-O-β-d-glucosyl cucurbitacin E. Nasturtium-reared larvae were insensitive to chlorogenic acid, which was deterrent to cabbage-reared larvae. Feeding by larvae reared on a wheat germ diet was not deterred by these compounds. The results indicate that dietary experience can extensively affect larval sensitivity to feeding deterrents and that cross habituation of larvae to deterrents occurs in response to certain chemical constituents of nasturtium and wheat germ diet. Digitoxin, however, proved to be an exception. Larvae reared on either nasturtium or wheat germ diet were as sensitive to digitoxin as those reared on cabbage. Previous results have shown that rejectionof nasturtium by cabbage-reared larvae is due to the presence of strong feeding deterrents in this plant. However, more than 50% of 2nd instar larvae reared from neonate on cabbage leaves treated with strophanthidin, cymarin, erysimoside, digitoxigenin and digitoxin accepted nasturtium as a food source. 2-O-β-d-glucosyl cucurbitacin E, 2-O-β-d-glucosyl cucurbitacin I and rutin were also active in causing larvae to feed on nasturtium. Thus dietary exposure to unrelated plant chemicals can profoundly affect insect acceptance of a plant that contains feeding deterrents.     

Cytosolic LOX overexpression in Arabidopsis enhances the attractiveness of parasitic wasps in response to herbivory and incidences of parasitism

Abstract

In response to herbivory by cabbage white butterfly (Pieris rapae) larvae, crucifer plants emit green leaf volatiles (GLVs) that attract Cotesia glomerata, a carnivorous natural enemy of the larvae. To artificially increase GLV production by crucifers, we created transgenic Arabidopsis that constitutively expressed cucumber cytosolic lipoxygenase (CsLOX2). Transgenic Arabidopsis (p35S::CsLOX2.6 and p35S::CsLOX2.14) infested with P. rapae larvae were more attractive to C. glomerata than wild type (wt) and produced more jasmonic acid. p35S::CsLOX2.14 had a higher incidence of parasitism of the larvae than did wt. Infested transgenic plants emitted more (Z)-3-hexenyl acetate, (E)-β-ocimene, (Z)-β-ocimene, and DMNT than infested wt plants; (Z)-3-hexenyl acetate is known to attract Cotesia wasps. Transforming cruciferous crops with cytosolic CsLOX2 could aid integrated pest management via tritrophic interactions.     

Response of the wasp (<Emphasis Type="Italic">Cotesia glomerata</Emphasis>) to larvae of the large white butterfly (<Emphasis Type="Italic">Pieris brassicae</Emphasis>)

The large white butterfly (Pieris brassicae L) first invaded northernmost Japan from Siberia around 1994, and after a few years, began to expand its range. The wasp, Cotesia glomerata (L) parasitizes larvae of the small white butterfly (Pieris rapae crucivora Boisduval), a usual host in the same geographic area. Some Pieris brassicae larvae in Hokkaido have been parasitized by Cotesia glomerata, but the parasitism rate of Pieris brassicae larvae tends to be lower than that of Pieris rapae. To examine the process of parasitizing Pieris brassicae larvae, we observed how the parasitoid wasp responded to the host larvae on damaged leaves. Cotesia glomerata females tended to avoid Pieris brassicae larvae, and even when female wasps inserted their ovipositors into Pieris brassicae larvae, none laid eggs. The parasitoids obtained from Pieris rapae larvae failed to parasitize Pieris brassicae during the host-acceptance step.     

Can herbivore‐induced plant volatiles inform predatory insect about the most suitable stage of its prey?

The leaf beetle Plagiodera versicolora (Coleoptera: Chrysomelidae) is a specialist herbivore, all of whose mobile stages feed on the leaves of salicaceous plants. Both the larval and adult stages of the ladybird Aiolocaria hexaspilota (Coleoptera: Coccinellidae) are dominant natural enemies of the larvae of the leaf beetle. To clarify the role of plant volatiles in prey‐finding behaviour of A. hexaspilota, the olfactory responses of the ladybird in a Y‐tube olfactometer are studied. The ladybird adults show no preference for willow plants Salix eriocarpa that are infested by leaf beetle adults (nonprey) over that for intact plants but move more to the willow plants infested by leaf beetle larvae (prey) than to intact plants. Moreover, ladybird larvae show no preference for willow plants infested by leaf beetle larvae or adults over intact plants. Using gas chromatography‐mass spectrometry, six volatile compounds are released in larger amounts in the headspace of willow plants infested by leaf beetle larvae than in the headspace of willow plants infested by leaf beetle adults. In addition, the total amount of volatiles emitted from willow plants that are either intact or infested by leaf beetle adults is much smaller than that from willow plants infested by leaf beetle larvae. These results indicate that volatiles from S. eriocarpa infested by P. versicolora inform A. hexaspilota adults about the presence of the most suitable stage of their prey, whereas A. hexaspilota larvae do not use such information.     

The parasitoidCotesia glomerata (Hymenoptera: Braconidae) discriminates between first and fifth larval instars of its hostPieris brassicae,on the basis of contact cues from frass,silk, and herbivore-damaged leaf tissue

Adult females of the larval parasitoidCotesia glomerata (L.) respond to chemical cues associated with feeding damage inflicted on cabbage plants by its host,Pieris brassicae (L.). The use of these infochemicals by the parasitoid during selection of the most suitable host instar was investigated. The parasitoid can successfully parasitize first-instar host larvae, while contacts with fifth-instar larvae are very risky since these caterpillars react to parasitization attempts by biting, spitting, and hitting, resulting in a high probability of the parasitoid being seriously injured or killed. Observations of the locomotor behavior of individual wasps on leaves with feeding damage inflicted by the first and the fifth larval instars and on host silk and frass showed that several cues affect the duration of searching by the parasitoids after reaching a leaf: cues on the margin of the feeding damage and cues in the host frass and silk. Whole frass, silk, and hexane extracts of frass obtained from first-instar elicited parasitoid's searching behavior significantly longer than frass, silk, and hexane extract of frass from the fifth instar. The results demonstrate thatC. glomerata can discriminate between first instars, which are more suitable hosts, and fifth instars ofP. brassicae without contacting the caterpillars, by exploiting instar-related cues.     

Differential selection of host plants by twoPieris species: the role of oviposition stimulants and deterrents

Oviposition responses ofPieris rapae L. andP. napi oleracea Harris to nine crucifers, one Capparidaceae and one Tropaeolaceae were directly compared under controlled conditions. Chemical fractions from these plants were also tested on both insects for the presence of oviposition stimulants or deterrents. The results showed that plant chemistry is a key factor in differential selection of potential hosts by thesePieris species. Some plant species were equally acceptable to bothPieris species. However,P. rapae preferred cabbage over most test plants whereasP. napi oleracea strongly preferred plant species that were avoided byP. rapae. The observed preferences were explained in most cases by the presence of stimulants and deterrents in extracts of the plants. The twoPieris species have apparently evolved differential sensitivities to the chemical stimuli that trigger or deter oviposition. The balance of positively and negatively interpreted sensory signals evoked by plant chemicals obviously plays an important role in acceptance or rejection of a plant by both species. The role of specific glucosinolates and differing structure-activity relationships is suggested.     

Influences of plant species on life history traits of Cotesia rubecula (Hymenoptera: Braconidae) and its host Pieris rapae (Lepidotera: Pieridae)

The relative suitability of four plants was studied for larvae of Pieris rapae L. and its parasitoid Cotesia rubecula (Marshall). For unparasitized P. rapae, pupal dry weight and egg-pupa growth rate were higher on cabbage, radish and nasturtium than on Indian hedge mustard. Larval developmental rate and size were greatest for C. rubecula when its host was feeding on nasturtium. Wasp survival was not affected by the host insect/plant combination in which the parasitoid developed. These results indicate that the plant on which host larvae feed is an important factor in development of the parasitoid.     

Effects of organic and conventional fertilizer treatments on host selection by the aphid parasitoid Diaeretiella rapae

The type and quantity of fertilizer supplied to a crop will differ between organic and conventional farming practices. Altering the type of fertilizer a plant is provided with can influence a plant’s foliar nitrogen levels, as well as the composition and concentration of defence compounds, such as glucosinolates. Many natural enemies of insect herbivores can respond to headspace volatiles emitted by the herbivores’ host plant in response to herbivory. We propose that manipulating fertilizer type may also influence the headspace volatile profiles of plants, and as a result, the tritrophic interactions that occur between plants, their insect pests and those pests’ natural enemies. Here, we investigate a tritrophic system consisting of cabbage plants, Brassica oleracea, a parasitoid, Diaeretiella rapae, and one of its hosts, the specialist cabbage aphid Brevicoryne brassicae. Brassica oleracea plants were provided with either no additional fertilization or one of three types of fertilizer: Nitram (ammonium nitrate), John Innes base or organic chicken manure. We investigated whether these changes would alter the rate of parasitism of aphids on those plants and whether any differences in parasitism could be explained by differences in attractivity of the plants to D. rapae or attack rate of aphids by D. rapae. In free‐choice experiments, there were significant differences in the percentage of B. brassicae parasitized by D. rapae between B. oleracea plants grown in different fertilizer treatments. In a series of dual‐choice Y‐tube olfactometry experiments, D. rapae females discriminated between B. brassicae‐infested and undamaged plants, but parasitoids did not discriminate between similarly infested plants grown in different fertilizer treatments. Correspondingly, in attack rate experiments, there were no differences in the rate that D. rapae attacked B. brassicae on B. oleracea plants grown in different fertilizer treatments. These findings are of direct relevance to sustainable and conventional farming practices.     

Foraging for solitarily and gregariously feeding caterpillars: A comparison of two related parasitoid species (Hymenoptera: Braconidae)

In the present study we apply a comparative approach, in combination with experimentation, to study behavior of two parasitoid species that attack caterpillar hosts with different feeding strategies (gregarious or solitary). In a semifield setup, consisting of clean cabbage plants and plants infested with one of two host species, the foraging behavior of the specialistCotesia rubecula, on obligate parasitoid of solitarily feedingPieris rapae larvae, was compared to that of the generalistCotesia glomerata, a polyphagous parasitoid of several Pieridae species (mainly the gregariously feedingPieris brassicae).Cotesia glomerata displayed equal propensity to search for and parasitize larvae of both host species. AlthoughC. glomerata exhibited a relatively plastic foraging behavior in that it searched differently under different host distribution conditions, its behavior seems more adapted to search for gregariously feeding hosts. Females exhibited a clear area-restricted search pattern and were more successful in finding the gregariously feeding caterpillars.Cotesia rubecula showed a higher propensity to search forP. rapae than forP. brassicae, i.e., females left the foraging setup significantly earlier when their natural hostP. rapae was not present.C. rubecula showed a more fixed foraging behavior, which seems adapted to foraging for solitarily feeding host larvae. In a setup with onlyP. rapae larvae, the foraging strategies of the two parasitoid species were quite similar. In a choice situationC. glomerata did not show a preference for one of the host species, whileCotesia rubecula showed a clear preference for its natural host species. The latter was shown by several behavioral parameters such as the number of first landings, allocation of search time, and percentage parasitization.     

Parasitoid-plant mutualism: parasitoid attack of herbivore increases plant reproduction

We tested whether a plant's life time seed production is increased by parasitization of herbivores in a tritrophic system, Arabidopsis thaliana (Brassicaceae) plants, Pieris rapae (Lepidoptera: Pieridae) caterpillars and the solitary endoparasitoid Cotesia rubecula (Hymenoptera: Braconidae). We established seed production for intact A. thaliana plants, plants that were mechanically damaged, plants fed upon by parasitized caterpillars and plants fed upon by unparasitized caterpillars. In the first experiment, with ecotype Landsberg (erecta mutant), herbivory by unparasitized P. rapae caterpillars resulted in a strongly reduced seed production compared to undamaged plants. In contrast, damage by P. rapae caterpillars that had been parasitized by C. rubecula did not result in a significant reduction in seed production. For the second experiment with the ecotype Columbia, the results were identical. Plants damaged by unparasitized caterpillars only produced seeds on regrown shoots. Seed production of plants that had been mechanically damaged was statistically similar to that of undamaged plants. Production of the first ripe siliques by plants fed upon by unparasitized caterpillars was delayed by 18–22 days for Landsberg and 9–10 days for Columbia. We conclude that parasitization of P. rapae by C. rubecula potentially confers a considerable fitness benefit for A. thaliana plants when compared to plants exposed to feeding damage by unparasitized P. rapae larvae. Plants that attract parasitoids and parasitoids that respond to herbivore-induced plant volatiles will both experience selective advantage, justifying the use of the term mutualism for this parasitoid-plant interaction. This type of mutualism is undoubtedly very common in nature.     

Electroantennogram responses to plant volatiles in two species of Pieris butterflies

Electroantennogram (EAG) responses were recorded from females of two related butterfly species, Pieris brassicae L. and P. rapae L. (Lepidoptera: Pieridae) to 23 volatiles of plant origin, 19 of which have been reported to occur in the headspace of their preferred host plants, Brassica and Sinapis species (Cruciferae). In both species, selective responses were observed and the most effective compounds were the 6-carbon fatty acid derivatives trans-hex-2-enal, hexan-1-ol and hexanal, which are common green leaf volatiles. Of the 6 isoprenoids tested, myrcene and geraniol were most effective. Of the 4 crucifer-specific compounds tested, phenylacetonitrile was a distinctly stronger stimulant than the three isothiocyanates in both species. The rank order of mean stimulating effectiveness of all compounds tested was strongly correlated between the two species. Statistical analysis of dose-response relationships for 6 compounds revealed significant differences between compounds. The rank order of effectiveness changed with dose. Exclusive exposure to Sinapis arvensis during larval life and young adulthood resulted in quantitative changes in EAG responses to several compounds in both species, although the overall rank order of effectiveness was strongly correlated between the groups reared on the two host plants.     

A comparative study of the egg-laying behaviour and larval development of Pieris rapae L. and P. brassicae L. on the same host plants

Summary Pieris rapae and P. brassicae feed on the same host plants and have synchronized seasons. P. brassicae, whose larvae are twice the size of P. rapae, lays eggs in clusters of 40–100 eggs whereas P. rapae lays single eggs. In this paper we examine how egg clustering may be advantageous for P. brassicae. The larval development of each species was studied, and found not to differ significantly. P. brassicae larvae were observed to migrate from their host plant after defoliating it. A comparison of the efficiency of host plant utilization by the two pierid species was undertaken by measuring the effect of larval feeding on the growth of their host plants (kale and brussel sprouts). The results show that egg clustering is advantageous for larval fitness in terms of host resource exploitation, and we suggest that P. brassicae is adapted for ovipositing on clumped vegetation, while P. rapae is selected for exploiting isolated plants.     

Induction of volatile emissions from ripening apple fruits infested with Cydia pomonella and the attraction of adult females

We studied the effect of herbivory by Cydia pomonella L. (Lepidoptera: Tortricidae) first instar larvae on the volatile emissions from immature apples collected at the end of August. Volatiles from infested, artificially damaged, and healthy fruit were collected using solid-phase microextraction and analysed using combined gas chromatography mass spectrometry. The results show that neonate C. pomonella larvae induce the release of E--ocimene from immature fruit compared to healthy or artificially damaged fruit. Seven further compounds, a terpene, an alcohol, four aldehydes, and a ketone, were consistently found in all treatments. There were no significant differences amongst these latter compounds, although the amount of the terpene E,E--farnesene was three to five times higher from the infested as compared to healthy fruits. Headspace volatiles were also collected on Tenax to test for behavioural responses to the volatiles. Volatile collections from infested fruit attracted mated adult females in a Y-tube olfactometer, whereas no preference was evinced for volatiles from healthy or artificially damaged fruits when compared to a solvent control.     

Impact of flowering buckwheat on Lepidopteran cabbage pests and their parasitoids at two spatial scales

We assessed the potential of annual buckwheat, Fagopyrum esculentum Moench, to lead to improved parasitism of lepidopteran cabbage pests over four years. Pest, parasitism, and hyperparasitism rates were monitored in replicated cabbage plots (12 × 20 m) with or without 3 m wide buckwheat borders from 2000 to 2003. Floral borders did not significantly increase egg, larval, or pupal densities of cabbage looper, Trichoplusia ni (Hübner), imported cabbageworm, Pieris rapae (L.), or diamondback moth, Plutella xylostella (L.). Buckwheat increased parasitism rates by Voria ruralis (Fallen) on T. ni larvae and Cotesia rubecula (Marshall) on P. rapaelarvae over four years. Parasitism by Diadegma insulare (Cresson) on P. xylostella larvae was higher in buckwheat than control plots in the first year, and parasitism by Euplectrus plathypenae (Howard) on T. ni larvae was lower in buckwheat than control plots in the second year. The hyperparasitoid Conura side (Walker) attacked D. insulare all four years, but buckwheat did not affect hyperparasitism rates. The effect of spatial scale on pest densities and parasitism in 2001 was evaluated by comparing plots separated at least 67 m (nearby) versus 800 m apart (isolated). T. ni pupae and P. rapae eggs and pupae were more abundant in plots in closer proximity, whereas P. xylostella densities did not vary by the spatial separation of plots. Tachinids and Pteromalus puparum (L.) attacked more P. rapae in nearby plots. E. plathypenae responded to the treatment × scale interaction, parasitizing more in control than buckwheat when plots were isolated but not when plots were nearby.