Divergence of defensive cucurbitacins in independent Cucurbita pepo domestication events leads to differences in specialist herbivore preference

Crop domestication and improvement often concurrently affect plant resistance to pests and production of secondary metabolites, creating challenges for isolating the ecological implications of selection for specific metabolites. Cucurbitacins are bitter triterpenoids with extreme phenotypic differences between Cucurbitaceae lineages, yet we lack integrated models of herbivore preference, cucurbitacin accumulation, and underlying genetic mechanisms. In Cucurbita pepo, we dissected the effect of cotyledon cucurbitacins on preference of a specialist insect pest (Acalymma vittatum) for multiple tissues, assessed genetic loci underlying cucurbitacin accumulation in diverse germplasm and a biparental F₂ population (from a cross between two independent domesticates), and characterized quantitative associations between gene expression and metabolites during seedling development. Acalymma vittatum affinity for cotyledons is mediated by cucurbitacins, but other traits contribute to whole-plant resistance. Cotyledon cucurbitacin accumulation was associated with population structure, and our genetic mapping identified a single locus, Bi-4, containing genes relevant to transport and regulation – not biosynthesis – that diverged between lineages. These candidate genes were expressed during seedling development, most prominently a putative secondary metabolite transporter. Taken together, these findings support the testable hypothesis that breeding for plant resistance to insects involves targeting genes for regulation and transport of defensive metabolites, in addition to core biosynthesis genes.     

Effects of host plant and cucurbitacin on growth of larval Diabrotica undecimpunctata howardi

The effects of host plant and dietary cucurbitacin on the growth of larval southern corn rootworm (SCR), Diabrotica undecimpunctata howardi Barber (Chrysomelidae: Luperini), were investigated. SCR were reared on four hosts: corn, Zea mays; peanuts, Arachis hypogaea; and two squash varieties, Cucurbita pepo cv. Ambassador (containing cucurbitacin D (0.08 mg g^–1 fr.wt.) = bitter), and C. pepo cv. Early Yellow Crookneck (lacking cucurbitacin = non-bitter). Larval growth was significantly greater on corn and peanuts than on either squash variety. After four weeks, adults had emerged from corn and peanut plants, while squash-reared larvae had not yet entered the pupal stage. There was no difference in larval growth on the two varieties of squash. Primary metabolite measurements showed no nutritional differences between the two squash varieties. Artificial diet experiments were used to test the effect of three concentrations of cucurbitacin D (0.0, 0.1, and 0.6 mg g^–1 diet) on growth of larval SCR. Larvae reared on diet containing 0.6 mg g^–1 cucurbitacin weighed significantly less than larvae reared on diet containing 0.1 mg g^–1 or no cucurbitacin after 10 d. No significant difference in growth was measured between the 0.1 mg g^–1 diet and the 0.0 mg g^–1 diet. Results are discussed relative to theories about the relationship between diabroticites and cucurbitacins.     

Consumptive and non‐consumptive effects of wolf spiders on cucumber beetles and cucumber plant damage

Biological control research often focuses on the ability of predators to reduce pest densities and protect crops through consumption. Less studied is their ability to protect crops by altering pest behaviour (non‐consumptive effects). Lab experiments were conducted to test predation rates of striped cucumber beetles (Acalymma vittatum; Coleoptera: Chrysomelidae) and spotted cucumber beetles (Diabrotica undecimpunctata howardi; Coleoptera: Chrysomelidae) by large (>10 mm) wolf spiders (Araneae: Lycosidae). Field experiments were conducted to examine how the physical presence and/or cues of spiders impact the behaviour and mortality of A. vittatum (specialist) and D. undecimpunctata (generalist) cucumber beetles as well as growth and damage of cucumber plants (Cucumis sativus; Cucurbitaceae). A. vittatum and D. undecimpunctata adults were added to caged cucumber plants without a spider, with spider cues only (spider removed before beetle inclusion), with spider only (spider introduced to plants immediately before beetle inclusion), and with spiders and their cues present (spiders introduced 24 hr in advance of beetle inclusion). A. vittatum responded to spider cues primarily by emigrating from plants. Contrarily, D. undecimpunctata did not display obvious responses, such as reduced feeding or increased emigration, to spider foraging and/or cues. Actively foraging lycosids increased A. vittatum mortality and reduced densities of D. undecimpunctata in the field when cucumber plants were flowering. This study highlights how non‐consumptive and consumptive effects can play a role in modifying pest populations, and how these effects can vary across species and plant growth stages.     

Evaluation of cucurbitacin-based gustatory stimulant to facilitate cucumber beetle (Coleoptera: Chrysomelidae) management with foliar insecticides in melons

The bitter plant-derived compounds cucurbitacins are known to stimulate feeding of adult cucumber beetles (Coleoptera: Chrysomelidae). A cucurbitacin-based gustatory stimulant applied as a flowable bait combined with either spinosad or carbaryl was compared with foliar sprays of spinosad and carbaryl for controlling two cucumber beetle species (Diabrotica undecimpunctata undecimpunctata Mannerheim and Acalymma trivittatum Mannerheim) in honeydew melons (Cucumis melo L.). Field studies were conducted on the University of California-Davis plant pathology farm in 2008 and 2009. Beetle densities after applications and fruit damage from beetle feeding were compared among treatments. In addition, beetle survival was compared within field cages placed over the treated foliage infested with beetles. Using all three measures of efficacy, we determined that the addition of cucurbitacin bait had no effect on the level of cucumber beetle control with carbaryl in either 2008 or 2009. In both years, spinosad did not significantly reduce cucumber beetle densities in either field cages or field plots and did not reduce fruit damage relative to the untreated control. The addition of the bait to spinosad did not improve its efficacy. A laboratory bioassay of the spinosad formulation used in the field showed it had significant lethal effects on adults of both cucumber beetle species. Results indicated that the bait formulation used did not improve cucumber beetle control but may benefit from the addition of floral attractants or using a different type of cucurbitacin.     

Plant exudates trigger leaf-trenching by cabbage loopers, Trichoplusia ni (Noctuidae)

Cabbage loopers, Trichoplusia ni, cut a narrow trench across leaves of plants that release exudate, then feed distal to the trench in an area of reduced exudation. The larvae do not normally trench plant species such as plantain, Plantago lanceolata, that lack exudate. To determine what cues elicit trenching, I reared larvae to the final instar on plantain, then applied test solutions to their mouthparts during feeding. Loopers that received latex from Lactuca serriola (Asteraceae) or phloem exudate from watermelon, Citrullus vulgaris (Cucurbitaceae), often responded by cutting a trench in plantain, even though these larvae had not previously encountered exudate nor previously trenched. Loopers that were allowed to trench and feed on L. serriola for 1 day prior to the assay subsequently cut trenches in plantain more frequently and in response to more fluids, including a viscous solution of polyethylene glycol and latex from a non-host, poinsettia (Euphorbia pulcherrima). Subsequent bioassays with larvae reared entirely on plantain tested whether bitter cucurbitacins or gelation are essential cues for trenching. Sap from non-bitter cucumber plants (Cucumis sativus) caused larvae to trench, showing that cucurbitacins are not required to induce trenching. Loopers also trenched after receiving cucumber sap that did not gel due to the addition of mercaptoethanol. An extract of sap lacking the proteins that cause gelation likewise triggered trenching. Further fractionation revealed that cucumber sap and also butternut squash sap (Cucurbita moschata) contain trenching stimulants that are small (molecular weight < 3,000) water-soluble molecules. Received: 13 January 1997 / Accepted: 6 June 1997     

Potential for habituation to a neem-based feeding deterrent in Japanese beetles, Popillia japonica

We tested the potential for the Japanese beetle, Popillia japonica Newman, to habituate to a neem-based feeding deterrent applied to foliage of linden, Tilia cordata L., a preferred host for the adults. Female beetles' consumption of control foliage versus foliage treated with either a low or high rate of neem insecticide, corresponding to 9 or 39 pm azadirachtin, respectively, was tested in a series of 4-h choice or no-choice tests over four successive days. In another experiment, females were conditioned for 22 h with either control foliage, leaves treated with the low rate, or a mixture of both treated and untreated leaves. Deterrence of either the low or high rate of neem to these beetles was then evaluated in choice tests with control foliage, as before. In choice tests, mean consumption of control foliage was always greater than for treated foliage, regardless of rate. There was, however, proportionately more feeding on foliage treated with the high rate upon successive exposures. In no-choice tests, beetles initially deterred by the low rate were not significantly deterred by that rate by the third and fourth days of the experiment. Finally, beetles conditioned by exposure to leaves treated with the low rate were not deterred by that rate in a subsequent choice test, although they were deterred by the higher rate. Despite these trends, we suggest that Japanese beetles' polyphagy and mobility probably would reduce the likelihood for habituation to neem-based feeding deterrents in the field.     

Mexican diabroticite beetles: I. Laboratory test on host breadth of Acalymma and Diabrotica spp.

Choice tests were conducted to determine relative degree of specialization of feeding behavior of 11 Mexican diabroticite species in the genera Acalymma and Diabrotica (Chrysomelidae: Luperini). Adult beetles were offered a choice between cotyledons of a non-bitter (not containing cucurbitacin) cucurbit (C. pepo L. var. Crookneck), corn (Zea mays L.), and beans (Phaseolus vulgaris L.). In a second assay a bitter (containing cucurbitacin) cucurbit (C. pepo L. var. Ambassador) was added to the array of plants offered. Neonates of two species of Acalymma and one species of Diabrotica were offered a choice between roots of a non-bitter and a bitter cucurbit, and between a bitter cucurbit and corn.Adult insects showed distinct preferences in the first assay. All Acalymma spp. tested accepted only the non-bitter cucurbit as host, whereas Diabrotica spp. preferred either the cucurbit or the noncucurbit hosts. When the bitter cucurbit was offered together with the other three hosts, all species changed their host choice and significantly preferred the bitter cucurbit. Neonates of all three species tested significantly preferred the bitter cucurbit roots over the non-bitter roots, and the corn roots over the bitter cucurbit.The observation that all Mexican diabroticite species tested left suitable hosts when bitter cucurbits were offered in a choice situation supported the hypothesis that the association between diabroticites and Cucurbitaceae is mediated by plant chemical compounds. For both, the Acalymma spp., which were found to be cucurbit specialists, as well as for the polyphagous Diabrotica spp., cucurbitacin B acted as a strong feeding arrestant which implies that the chemical mediation of this interaction might be an evolutionary conservative trait within the tribe.     

A Male-Produced Aggregation Pheromone Facilitating Acalymma vittatum [F.] (Coleoptera: Chrysomelidae) Early-Season Host Plant Colonization

We conducted field studies to investigate the involvement of volatile cues in early-season host plant colonization by striped cucumber beetle, Acalymma vittatum (F.) (Coleoptera: Chrysomelidae). Wind-directed traps were baited with male or female A. vittatum, potted cucumber (Cucumis sativus) seedlings that were of near-isogenic lines which either contained or lacked cucurbitacin, or combinations of male or female A. vittatum feeding on one or the other cucumber variety. We found no response to undamaged plants of either cucumber variety or plants that were actively being fed upon by A. vittatum females, whereas the response to volatiles associated with male A. vittatum was strong. Both male and female conspecifics and totals of up to sevenfold the number of males in the trap lures were caught overnight. Feeding males attracted more than double the number of conspecifics that responded to nonfeeding males. Active consumption of cucurbitacin in the plant on which the males were feeding, however, had no effect on attraction. A shift in sex ratio from a male to a female bias during field colonization season also supports the hypothesis that host finding is initiated by pioneer males. The importance of this aggregation pheromone in early-season host plant colonization and the evolutionary and adaptive significance of this pheromone are discussed.     

Pathogen-activated induced resistance of cucumber: response of arthropod herbivores to systemically protected leaves

Summary Restricted (non-systemic) inoculation of cucurbits, green bean, tobacco, and other plants with certain viruses, bacteria, or fungi has been shown to induce persistent, systemic resistance to a wide range of diseases caused by diverse pathogens. The non-specificity of this response has fueled speculation that it may also affect plant suitability for arthropod herbivores, and there is limited evidence, mainly from work with tobacco, which suggests that this may indeed occur. Young cucumber plants were immunized by restricted infection of a lower leaf with tobacco necrosis virus (TNV), and upper leaves were later challenged with anthracnose fungus, Colletotrichum lagenarium, to confirm induction of systemic resistance to a different pathogen. The response of arthropod herbivores was simultaneously measured on non-infected, systemically protected leaves of the same plants. As has been reported before, immunization with TNV gave a high degree of protection from C. lagenarium, reducing the number of lesions and the area of fungal necrosis by 65–93%. However, there was no systemic effect on population growth of twospotted spider mites, Tetranychus urticae Koch, on upper leaves, nor did restricted TNV infection of leaf tissue on one side of the mid-vein systemically affect mite performance on the opposite, virus-free side of the leaf. Similarly, there were no effects on growth rate, pupal weight, or survival when fall armyworm larvae were reared on systemically protected leaves from induced plants. In free-choice tests, greenhouse whiteflies oviposited indiscriminately on induced and control plants. Feeding preference of fall armyworms was variable, but striped cucumber beetles consistently fed more on induced than on control plants. There was no increase in levels of cucurbitacins, however, in systemically-protected foliage of induced plants. These findings indicate that pathogen-activated induced resistance of cucumber is unlikely to provide significant protection from herbivory. The mechanisms and specificity of induced resistance in cucurbits apparently differ in response to induction by pathogens or herbivores.     

Plant stress and insect performance: cottonwood,ozone and a leaf beetle

Summary Leaf area consumption rates, development rates, survivorship, and fecundity of the imported willow leaf beetle (Plagiodera versicolora Laich) were examined on two clones of eastern cottonwood which were previously exposed to ozone or charcoal-filtered air. P. versicolora consumed more ozone treated foliage, but were more fecund when reared on charcoal-filtered air treated plants. Beetle development rates and survivorship were not significantly different on treated and control cottonwoods. We concluded that: 1) Ozone fumigation of cottonwood reduced foliage quality, and the reproductive success and overall performance of P. versicolora. 2) increased foliage consumption by beetles was probably a mechanism compensating for decreases in foliage quality. 3) Reductions in beetle fecundity were due to an initial reduction in oviposition rates. 4) Beetle feeding preference did not correlate with the suitability of foliage for beetle performance. These results are discussed in relation to the impact of air pollution on plant-insect interactions.     

Cucurbitacine als stathmokinetische Wirkstoffe

Summary A synergism of colchicine and different cucurbitacins has been shown, to be a cause in the formation of ball-metaphases in Bryonia species. The bitter principles cucurbitacin L and I, dihydrocucurbitacin B, and tetrahydrocucurbitacin I, which were examined more closely with respect to their karyological effects, were shown to give rise to C-metaphases in Hordeum and Vicia at concentrations of about 0.016 per cent. On the one hand the number of mitotic stages is reduced, and on the other hand there is an increase of metaphase stages. Higher concentrations lead to pycnotic degenerations of nuclei and are toxic. When colchicine is applied together with cucurbitacins, ball-metaphases result. This could be demonstrated in plants lacking colchicine and cucurbitacins (Hordeum) when both substances were applied at the same time. Ball-metaphases also appear in plants containing cucurbitacins (Bryonia alba, B. dioica, Citrullus colocythis, Iberis amara) when they are treated with colchicine.     

Convergent evolution of cucurbitacin feeding in spatially isolated rootworm taxa (Coleoptera: Chrysomelidae; Galerucinae,Luperini)

Historically, chemical ecologists assumed that cucurbitacin feeding and sequestration in rootworm leaf beetles is a remnant of an ancient association between the Luperini (Coleoptera: Chrysomelidae; Galerucinae) and Cucurbitaceae (ancestral host hypothesis). Under this premise, rootworms that do not develop on cucurbits but undergo pharmacophagous forays for cucurbitacins are thought to do so to supplement novel host diets that lack these bitter compounds. The ancestral host hypothesis is supported from studies of pyrrolizidine alkaloid pharmacophagy in Lepidoptera but has not been subjected to phylogenetic analysis within the Luperini. New evidence that this feeding behavior is better correlated with an adult affinity for pollen than with larval host offers the possibility that Old and New World rootworm species with an affinity for cucurbitacins converged on this behavior through apomorphic taste receptor modifications (loose receptor hypothesis). Here we test the monophyly of cucurbitacin feeding within the Luperini by using nuclear and mitochondrial sequence data to infer phylogenetic relationships among 49 taxa representing tribes of the Galerucinae and subtribes of the Luperini. The resulting phylogenetic hypothesis is mostly concordant with existing tribal and subtribal delineations within the Subfamily Galerucinae sensu stricto (Galerucinae not including the flea beetles). The establishment of ancestry among the subtribes of the Luperini refutes the monophyly of cucurbitacin feeding and cucurbit specialization, with the New World Diabroticina being paraphyletic to the Old World Aulacophorina and cosmopolitan Luperina. These data unambiguously support the convergent evolution of cucurbitacin feeding in rootworms and are inconsistent with the ancestral host hypothesis.     

Male traits under cryptic female choice in the spotted cucumber beetle (Coleoptera: Chrysomelidae)

Males of the spotted cucumber beetle (Diabrotica undecimpunctata howardi) rhythmically stroke females with their antennae during copulation. Males that stroke quickly have a higher probabilityof being accepted as a mate. We determined (1) the mechanismby which females prevent unattractive males from passing spermatophores,(2) whether antennal stroking signals to females the likelihoodof receiving a nuptial gift, and (3) if other male traits inaddition to stroking are subjected to sexual selection fromfemale preference. Dissections of pairs flash-frozen in copuladuring and after antennal stroking showed musculature that,when contracted, folded the vaginal duct leading to the female'sbursa copulatrix in a way that prevented complete penetrationby the aedeagus. These muscles were always contracted whilemales were stroking and always relaxed after stroking had ceased.Males accepted as mates did not differ from males that failedto pass a spermatophore in either absolute or relative bodyweight, aedeagus length, or the amount of cucurbitacins (potentialnuptial gifts) sequestered in their spermatophores. Although99% of the beetles that came to cucurbitacin-rich Cucurbitafruits in the field were males, males that had sequestered cucurbitacins did not stroke females faster than males withno cucurbitacins, and fast-stroking males were not more likelyto find and sequester cucurbitacins than were males that strokedmore slowly. Males with a cucurbitacin slurry painted on theirantennae had no mating advantage over controls. We concludethat females discriminate among males after copulation hasbegun on the basis of antennal stroking displays (or some traitcorrelated with stroking speed) that males perform to enticefemales to relax their bursal sphincter.     

An ecological cost of plant defence: attractiveness of bitter cucumber plants to natural enemies of herbivores

Plants produce defences that act directly on herbivores and indirectly via the attraction of natural enemies of herbivores. We examined the pleiotropic effects of direct chemical defence production on indirect defence employing near‐isogenic varieties of cucumber plants (Cucumis sativus) that differ qualitatively in the production of terpenoid cucurbitacins, the most bitter compounds known. In release–recapture experiments conducted in greenhouse common gardens, blind predatory mites were attracted to plants infested by herbivorous mites. Infested sweet plants (lacking cucurbitacins), however, attracted 37% more predatory mites than infested bitter plants (that produce constitutive and inducible cucurbitacins). Analysis of the headspace of plants revealed that production of cucurbitacins was genetically correlated with large increases in the qualitative and quantitative spectrum of volatile compounds produced by plants, including induced production of (E )‐β‐ocimene (3E )‐4,8‐dimethyl‐1,3,7‐nonatriene, (E,E)‐α‐farnesene, and methyl salicylate, all known to be attractants of predators. Nevertheless, plants that produced cucurbitacins were less attractive to predatory mites than plants that lacked cucurbitacins and predators were also half as fecund on these bitter plants. Thus, we provide novel evidence for an ecological trade‐off between direct and indirect plant defence. This cost of defence is mediated by the effects of cucurbitacins on predator fecundity and potentially by the production of volatile compounds that may be repellent to predators.     

Insect feeding on cucumber mediated by rhizobacteria-induced plant resistance

Select strains of plant growth-promoting rhizobacteria (PGPR) were evaluated in greenhouse experiments with cucumber for induction of resistance against cucumber beetle (Diabrotica undecimpunctata howardi Barber) feeding and the beetle-transmitted cucurbit wilt disease. When beetles were given a choice between PGPR-treated and nontreated cucumber, their feeding on stems and cotyledons and the severity of wilt symptoms were significantly lower on PGPR-treated plants. HPLC analysis demonstrated that cotyledons from PGPR-treated plants contained significantly lower concentrations of the cucumber beetle feeding stimulant cucurbitacin than nontreated plants. These results suggest that a mechanism for PGPR-induced resistance against cucumber beetle feeding may involve a change in the metabolic pathway for cucurbitacin synthesis.     

Cucurbitacins and predation of the spotted cucumber beetle, Diabrotica undecimpunctata howardi

Two hypotheses about the ralationship of diabroticina beetles and plants in the family Cucurbitaceae are tested: (1) evolution of sensory receptors for cucurbitacins by diabroticina beetles was in part due to the predator protection offered by ingestion of these compounds, and (2) commercial varieties of cucurbitacin-producing cucumber offer Diabrotica undecimpunctata howardi Barber chemical protection from some potential sources of natural controlSpotted cucumber beetles fed either Marketmore 70 cucumber which contains cucurbitacin-C or Marketmore 72 which totally lacks cucurbitacin were presented to four species of vertebrate predators that commonly occur in the summer and/or winter habitats of D. u. howardi: Bufo americanus, B. fowleri, Peromyscus maniculatus, Colinus virginianus. None of the four species were significantly deterred from preying on beetles that had eaten Marketmore 70 cucumber. These results do not support either of the two hypotheses. The limitations of these negative results as evidence for refutation of the first hypothesis are discussed.

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Résumé Deux hypothèses concernant les relations entre les Diabroticina et les Cucurbitaceae ont été examinées: (1) l'évolution chez les Diabroticina des récepteurs pour les Cucurbitacines est due en partie à la protection contre les prédateurs apportée par la consommation de ces produits (2) les variétés modernes de concombres produisant de la cucurbitacine fournissent à D. undecimpunctata howardi Barber une protection chimique contre quelques sources potentielles de lutte biologique.Des D. undecimpunctata howardi, ayant consommé soit des concombres Marketmore 70 contenant de la cucurbitacine C, soit des concombres Marketmore 72 qui ont totalement perdu leur cucurbitacine, ont été présentés à quatre espèces de vertébrés prédateurs, qui s'observent fréquemment dans les habitats d'été ou d'hiver de D. undecimpunctata howardi soient Bufo americanus, B. fowleri, Peromyscus maniculatus, Colinus virginianus. Aucune de ces quatre espèces n'est significativement dissuadée d'attaquer les coléoptères qui on consommé les concombres Marketmore 70. Ces résultats n'étayent aucune des deux hypothèses. La discussion porte sur les limites de ces résultats négatifs comme preuve de la réfutation de l'hypothèse 1.

     

Plant-mediated interactions between insects and a fungal plant pathogen and the role of plant chemical responses to infection

Diverse organisms simultaneously exploit plants in nature, but most studies do not examine multiple types of exploiters like phytophagous insects and fungal, bacterial, and viral plant pathogens. This study examined patterns of induction of antipathogenic peroxidase enzymes and phenolics after infection by the cucurbit scab fungus, Cladosporium cucumerinum, and then determined if induction mediated ecological effects on Colletotrichum orbiculare, another fungal pathogen, and two insect herbivores, spotted cucumber beetles, and melon aphids. Peroxidase induction occurred in inoculated, `local,' symptom-bearing leaves 3 days after inoculation, and in `systemic,' symptom-free leaves on the same plants 1 day later. Phenolics were elevated in systemic but not in local leaves 3 days after inoculation. Detached systemic leaves from plants inoculated with C. cucumerinum developed significantly fewer and smaller lesions after challenge with C. orbiculare. Spotted cucumber beetles did not show consistently significant preferences for infected versus control leaf disks in comparisons using local or systemic leaves, but trends differed significantly between leaf positions. In no-choice tests, beetles removed more leaf area from local but not from systemic infected leaves compared to control leaves, and melon aphid reproduction was enhanced on local infected leaves. In the field, cucumber beetle and melon aphid densities did not differ between infected and control plants. Antipathogenic plant chemical responses did not predict reduced herbivory by insects. Other changes in metabolism may explain the positive direction and spatially dependent nature of plant-mediated interactions between pathogens and insects in this system. Received: 28 September 1997 / Accepted: 9 February 1998     

Artificial diet and rearing methods for Thanasimus dubius (Coleoptera: Cleridae), a predator of bark beetles (Coleoptera: Scolytidae)

Clerid beetles are common natural enemies of bark beetles, and could potentially be used as biological control agents if they could be reared in sufficient numbers. We developed an artificial diet devoid of insect components for rearing Thanasimus dubius (Fabricius), a clerid that attacks several economically important bark beetles in eastern North America. We reared larvae of this predator using the artificial diet, and then used either natural or factitious prey to feed the adults so produced. Two different methods of presenting the diet were also examined. We then compared the performance of T. dubius reared on the artificial diet with newly-emerged wild individuals collected from the field. Our results suggest that adult predators reared on the diet are near in quality to wild ones, and high R₀ values can be obtained. No difference in prey preference was found between wild and diet-reared individuals after five generations in the laboratory. Sufficient numbers of predators could be generated using these techniques to permit limited field trials of augmentative biological control.     

Development and feeding of <Emphasis Type="Italic">Tetraphleps galchanoides</Emphasis>, a predator of the hemlock woolly adelgid

Tetraphleps galchanoides Ghauri (Hemiptera: Anthocoridae) nymphs were collected from hemlock woolly adelgid (HWA) Adelges tsugae Annand (Hemiptera: Adelgidae) infested Tsuga sp. in Baoxing, Sichuan, China. First and second stage nymphs collected from foliage shipped from China; were reared to adults and tested for feeding rates and host preferences. They were reared at 5, 8, 12, and 15 ± 1 °C from November to December, January to March, April, and May to June, respectively, in the quarantine laboratory at Virginia Polytechnic Institute and State University. At 8 °C, development time was 15, 20, and 40 days for the N-III, IV, and V nymphal stages, respectively. Adult males lived 83 days with a range of 21–147 days. A single adult female lived for 21 days. At 5 °C, second stage T. galchanoides nymphs consumed 0.8 HWA nymphs per day, and 2.0 HWA nymphs per day at the N-V stage. At 8 °C, consumption of HWA nymphs ranged from 1.3 to 3.4 nymphs per day for the N-III to N-V stages, respectively. Adult T. galchanoides consumed more HWA eggs than HWA adults, pine bark adelgid (PBA) Pineus strobi (Hartig) (Hemiptera: Adelgidae) adults, and eggs in no-choice tests. In choice tests with HWA eggs and PBA eggs, more HWA eggs were eaten. Adult and nymph body measurements are presented for determination of nymphal instars.     

Glandular trichomes ofSolanum berthaultii alter host preference of the Colorado potato Beetle,Leptinotarsa decemlineata

Choice and no-choice studies were conducted to determine how the glandular trichomes of the wild potato,Solanum berthaultii Hawkes, affect host preference of the Colorado potato beetle,Leptinotarsa decemlineata (Say). Given a feeding choice betweenS. tuberosum andS. berthaultii, larvae and adults preferred the foliage ofS. tuberosum, but adults were more discriminating. When foliage ofS. berthaultii was appressed toS. tuberosum leaflets, fewer adults fed on the appressed leaflets. When given a choice between ‘trichome-intact’ and ‘trichome-removed’S. berthaultii foliage, adults preferred to feed on the latter. The preference for ‘trichome-removed’ foliage and the percent of adults initiating feeding, increased with the degree of trichome removal. These studies provide evidence that the resistance ofS. berthaultii is associated with feeding deterrents localized in the glandular trichomes, thatS. berthaultii possesses more than one mechanism of resistance to the Colorado potato beetle, and that the expression of resistance is dependent on the developmental stage of the insect.