Effect of soybean phytoalexins on the herbivorous insects mexican bean beetle and soybean looper

Effects of soybean phytoalexins on the feeding of the soybean looper and Mexican bean beetle were investigated to test the hypothesis that phytoalexins might be a defense mechanism of plants against insects as well as against pathogens. Short-term behavioral responses to the phytoalexins were analyzed using dual-choice tests with phytoalexin-rich and phytoalexin-poor (control) tissues. Phytoalexin production was elicited with ultraviolet radiation. Results from the dual-choice tests indicated that 6th instar soybean looper larvae fed equally on the control and phytoalexin-rich tissues. Feeding by adult and 4th instar Mexican bean beetles, however, was strongly deterred by the phytoalexins as evidenced by “single-bite” mandible scars on the phytoalexin-rich cotyledon discs. Nutritional effects of the isoflavonoid phytoalexin glyceollin on early instar soybean looper larvae were tested by incorporating the phytoalexin into an artificial medium at a level of 1% dry weight (0.15% fresh weight). The larvae were reared for 7 days from emergence on diets of control and glyceollin-containing media. Although survival on the glyceollin diets was initially less than on the control diets, under the experimental conditions glyceollin had no significant effect on the growth, development, or subsequent survival of the larvae. Efficiency of food utilization (ECI) was reduced, indicating that the phytoalexins may be a mild digestibility-reducing factor for the loopers. Implications of the results for host-plant resistance are discussed.     

Effect of DIMBOA,an aphid resistance factor in wheat,on the aphid predatorEriopis connexa Germar (Coleoptera: Coccinellidae)

DIMBOA (2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one), a secondary metabolite found in cereal extracts, confers resistance in wheat to aphids. Its effect on beneficial organisms was tested on larvae of the aphid predatorEriopis connexa Germar. Larvae were fed until pupation on artificial diets to which different concentrations of DIMBOA (2–200g/g diet) were added, as well as on aphids that had been feeding on wheat seedlings with different DIMBOA levels (140–440 g/g fresh tissue). In diets, the effect of DIMBOA was greatest on survival of third-instar larvae and on the duration of the second and fourth instars. When aphids were provided as food, those that had fed on a wheat cultivar with an intermediate DIMBOA level led to a significantly longer larval duration in the predator than did those that fed on either low or high DIMBOA cultivars. Shortest predator development times were obtained with aphid prey that had fed on high DIMBOA seedlings. Higher DIMBOA levels in the plant appear to reduce aphid feeding rates (and rates of DIMBOA ingestion), decreasing aphid survival and minimizing the effect of the toxin on the predator.On leave from Departamento de Entomologia, Universidad Nacional Agraria La Molina, Apartado aéreo 456, Lima, Perú.     

Ethanol and Ambrosia Beetles in Douglas Fir Logs Exposed or Protected from Rain

Logs from the base of Douglas fir (Pseudotsuga menziesii) trees cut in October 1993 were randomly assigned to one of three treatment groups: (1) wet logs—cut from the fallen tree and left exposed to rain, (2) dry logs—cut from the fallen tree, placed on blocks, and protected from rain under a plastic tent, and (3) crown logs—left attached to the fallen tree with its branches intact and exposed to rain. The following May, ethanol concentrations were highest in the phloem and sapwood of wet logs (0.24 and 0.35 mol/g fresh wt, respectively). Ethanol concentrations in tissues from dry and crown logs were similar to each other (ranging from 0.002 to 0.03 mol/g fresh wt), but were significantly lower than in wet logs. It appears that rain absorbed by the outer bark of wet logs creates a barrier to gas exchange between living tissues and the atmosphere, which facilitates the development of hypoxic conditions necessary for ethanol synthesis and accumulation. Branches on crown logs exposed to rain help maintain low ethanol concentrations in the log tissues; we discuss several potential mechanisms to explain this response. By early September, the densities of Gnathothrichus spp. gallery entrance holes were high on wet logs (21.5/m²) and low on dry (2.5/m²) and crown logs (5.8/m²), indicating their preference for logs with higher ethanol concentrations. Protecting logs from rain will significantly reduce ethanol concentrations and the density of ambrosia beetle galleries. Leaving branches attached to logs will produce similar results, but its effectiveness may vary depending on the environmental conditions. Host selection by secondary scolytid beetles that use ethanol as a kairomone can be manipulated and possibly managed by controlling the production of ethanol in the host resource.     

Effect of fungal metabolite peramine and analogs on feeding and development of argentine stem weevil (Listronotus bonariensis)

Peramine, a pyrrolopyrazine alkaloid produced by the fungal endophyte of perennial ryegrassAcremonium lolii, deterred the feeding of both adults and larvae of the graminacious herbivore, the Argentine stem weevil (Listronotus bonariensis), at 0.1 g/g and 10 g/g, respectively. In a no-choice test fewer stem weevil larvae fed and developed on diet containing as little as 2 g/g peramine. The proportion of larvae which did not develop beyond the first instar was higher on diet containing peramine and appeared to be due to a higher proportion of larvae which did not feed. For larvae which fed on the peramine-containing diet, feeding scores and times to pupation were not significantly different from those of controls. A number of simple peramine analogues showed feeding-deterrent activity against adult weevils, indicating the importance of the pyrrolopyrazine ring system of peramine in determining feeding-deterrent activity.     

Larval exposure to oviposition deterrents alters subsequent oviposition behavior in generalist,Trichoplusia ni and specialist,Plutella xylostella moths

The present study was undertaken to determine the effects of larval feeding experience on subsequent oviposition behavior of the resulting moths. Larvae of the cabbage looper (Trichoplusia ni, Noctuidae) and the diamondback moth (Plutella xylostella, Plutellidae) were exposed to the phenylpropanoid allelochemical trans-anethole (at 100 ppm fw in artificial diet) or the limonoid allelochemical toosendanin (10 ppm sprayed on cabbage leaves). Both compounds had been shown to deter oviposition in naïve moths in previous choice tests. Moths developing from experienced larvae (both sexes) showed a decrease in oviposition deterrence response when given a choice between control and treated leaves, unlike naïve moths. This phenomenon, analogous to habituation to feeding deterrents in lepidopteran larva, occurred irrespective of duration of feeding on the deterrent compound. We also observed that F₁larvae resulting from experienced moths (previously exposed to toosendanin as larvae) grew as well on toosendanin-treated foliage as on control foliage. In contrast, growth of F₁larvae from naïve moths was significantly impaired by toosendanin. These results demonstrate that host-selection behavior in cabbage looper (a generalist) and diamondback moth (a specialist) may be shaped by feeding experience according to Hopkins' Host Selection Principle in addition to chemical legacy.     

α-Tocopherol alteration of soybean antiherbivory toTrichoplusia ni larvae

The antioxidant vitamin E, -tocopherol, was tested as a candidate elicitor of alterable antiherbivory in soybean plants against cabbage looper larvae. Although a nonspecific antioxidant, vitamin E proved elicitory to the involved sulfhydryl-dependent receptor-energy transducer protein in soybean plasma membrane. Effects of -tocopherol were dependent on dosage, time, and space in the plant. The observed elicited effects were all decreases in herbivory. The best negative phytochemical correlate of looper feeding was the percentage of increased total HPLC peak area of extractables from elicited as compared to nonelicited leaves. Some specific compounds, e.g., glyceollins, were quantitatively major components of the total profile of secondary metabolites.     

Behavioral and physiological responses of cabbage looper,Trichoplusia ni (Hübner), to steam distillates from resistant versus susceptible soybean plants

Soybean plant volatiles, extracted as steam distillates, significantly affected the behavior and biology of the cabbage looper,Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae). Distillates from the susceptible Davis variety attractedT. ni larvae and female adults, whereas those from resistant PI 227687 plants repelled them. When mixed in an artificial diet, steam distillates from PI 227687 plants caused mortality of first-instar larvae. Adults emerging from pupae topically treated with 5 g of such PI 227687 extractable showed developmental abnormalities. Larval feeding was significantly less on Davis leaves treated with PI 227687 volatiles as compared to solvent (acetone) or such Davis extractables. However, Davis volatiles on PI 227687 leaves did not increase larval feeding. HPLC analyses of steam distillates from susceptible Davis versus resistant PI 227687 indicated differences.     

Sequestration of Host Plant Glucosinolates in the Defensive Hemolymph of the Sawfly Athalia rosae

Interactions between insects and glucosinolate-containing plant species have been investigated for a long time. Although the glucosinolate–myrosinase system is believed to act as a defense mechanism against generalist herbivores and fungi, several specialist insects use these secondary metabolites for host plant finding and acceptance and can handle them physiologically. However, sequestration of glucosinolates in specialist herbivores has been less well studied. Larvae of the turnip sawfly Athalia rosae feed on several glucosinolate-containing plant species. When larvae are disturbed by antagonists, they release one or more small droplets of hemolymph from their integument. This reflex bleeding is used as a defense mechanism. Specific glucosinolate analysis, by conversion to desulfoglucosinolates and analysis of these by high-performance liquid chromatography coupled to diode array UV spectroscopy and mass spectrometry, revealed that larvae incorporate and concentrate the plant's characteristic glucosinolates from their hosts. Extracts of larvae that were reared on Sinapis alba contained sinalbin, even when the larvae were first starved for 22 hr and, thus, had empty guts. Hemolymph was analyzed from larvae that were reared on either S. alba, Brassica nigra, or Barbarea stricta. Leaves were analyzed from the same plants the larvae had fed on. Sinalbin (from S. alba), sinigrin (B. nigra), or glucobarbarin and glucobrassicin (B. stricta) were present in leaves in concentrations less than 1 mol/g fresh weight, while the same glucosinolates could be detected in the larvae's hemolymph in concentrations between 10 and 31 mol/g fresh weight, except that glucobrassicin was present only as a trace. In larval feces, only trace amounts of glucosinolates (sinalbin and sinigrin) could be detected. The glucosinolates were likewise found in freshly emerged adults, showing that the sequestered phytochemicals were transferred through the pupal stage.     

Dietary influences on terpenoids sequestered by the biological control agent Oxyops vitiosa: effect of plant volatiles from different Melaleuca quinquenervia chemotypes and laboratory host species

The weevil Oxyops vitiosa is an Australian species imported to Florida, USA, for the biological control of the invasive species Melaleuca quinquenervia. The larvae of this species feed on the leaves of their host and produce a shiny orange secretion that covers their integument. Previous results indicated that a major component of this secretion, viridiflorol, is sequestered from the host plant and repels a generalist predator, the red imported fire ant, Solenopsis invicta. When the larvae fed on a different chemotype of M. quinquenervia, which lacked viridiflorol but was rich in a different sesquiterpene, (E)-nerolidol, similar protection occurred. Solvent washes of these larvae indicated that (E)-nerolidol was sequestered from M. quinquenervia leaves and repelled S. invicta workers when applied to dog food baits at physiological concentrations (17.5, 35.0, and 52.5 g/mg). Additionally, -caryophyllene also repelled S. invicta workers when applied to dog food baits at concentrations that approximated those in the O. vitiosa larval secretions (3.5 and 35 g/mg). When the O. vitiosa larvae were fed leaves from laboratory hosts (nonfield hosts), similar repellent activity was found. This activity was traced to several of the same compounds (e.g., 1,8-cineole, viridiflorol) found to be active in their field host M. quinquenervia. These weevil larvae are opportunistic, sequestering the primary terpenoids in their host leaves that confer antipredator activity.     

Are Insect-Synthesized Retronecine Esters (Creatonotines) the Precursors of the Male Courtship Pheromone in the Arctiid Moth Estigmene acrea?

The pyrrolizidine alkaloid (PA) profiles were determined for adults of the polyphagous arctiid Estigmene acrea, which as larvae had fed on artificial diet supplemented with Crotalaria-pumila powder with known concentrations of PAs. The larvae always had a free choice between alkaloid-containing and plain diets. The alkaloid profiles of adults revealed a striking sexual dimorphism. Both sexes contained macrocyclic PAs of the monocrotaline type sequestered from the diet and, in addition, a substantial proportion of supinidine and retronecine esters synthesized by the insects from necine bases derived from the dietary alkaloids and necic acids of insect origin. These insect alkaloids accounted for 35% and 55% of total PAs in males and females, respectively. The difference was that in females the retronecine esters (creatonotines) made up 58 g (43% of total PAs), while males contained a fivefold lower proportion, 12 g (13%). Four of the ten male individuals analyzed were found devoid of creatonotines. Based on the experimental data in combination with evidence from the literature, it is suggested that the creatonotines are direct pheromone precursors in E. acrea. It is hypothesized that this may represent a general mechanism of hydroxydanaidal formation from diverse macrocyclic PAs in arctiids.     

Alleviation of α-tomatine-induced toxicity to the parasitoid,Hyposoter exiguae,by phytosterols in the diet of the host,Heliothis zea

The solitary, endoparasitic ichneumonid,Hyposoter exiguae (Viereck) is a parasitoid of the tomato fruitworm,Heliothis zea (Boddie). However, the parasitoid is deleteriously affected by the tomato glycoalkaloid, -tomatine, ingested from hosts (H. zea) fed on artificial diets or semipurified extracts of tomato plants that contained -tomatine. -Tomatine causes prolonged larval development; disruption or prevention of pupal eclosion; deformation of antennal, abdominal, and genital structures; and reduction in adult weight and longevity of the parasitoid. These toxic effects are exacerbated when the dietary dose of -tomatine is increased from 12 mol to 20 mol/g dry wt of diet. However, the toxicity of -tomatine is attenuated in parasitoids reared from hosts fed on artificial diets that contain equimolar or supramolar amounts of 3-OH-sterols admixed with -tomatine. Further, the toxicity of extracts from the foliage of different cultivars of tomatoes toH. exiguae is contingent upon the composite levels of -tomatine and total phytosterol (free and esterified). Cultivars with a low total sterol-tomatine ratio are more toxic toH. exiguae. The possible mode of action of -tomatine toxicity toH. exiguae by disruption of sterol metabolism is discussed.     

Sex pheromone components of the blueberry leaftier mothCroesia curvalana kearfott (Lepidoptera: Tortricidae)

The GC-MS and GC-EAD analyses of sex-gland components and sex-gland-released volatiles have identified (E)- and (Z)-11-tetradecenal (9010) and (E)- and (Z)-11-tetradecenyl acetate (8515) as components of the sex pheromone chemical communication system ofCroesia curvalana (Kearfott). The aldehyde-to-acetate ratio of 91 was found in the gland-released components. This blend at a 0.1–1 g/day release rate from PVC lures was shown to be as effective as virgin females in the trapping of maleC. curvalana.     

Interaction of nuclear polyhedrosis virus with catechols: Potential incompatibility for host-plant resistance against noctuid larvae

Two major orthodihydroxy phenolics ofLycopersicon esculentum, rutin and chlorogenic acid, have previously been identified as potential sources of host-plant resistance against the tomato fruitwormHeliothis zea. We report here the possible incompatibility of these chemically based resistance factors with viral control ofH, zea. We have found that both rutin and chlorogenic acid significantly inhibited the infectivity of nuclear polyhedrosis viruses. Chlorogenic acid, when added to tissue culture medium containing TN-368 ovarian cells, inhibited the infectivity of a multiply embedded virus (AcMNPV) by over 86%. Rutin or chlorogenic acid, when fed toH. zea, inhibited the infectivity of a singly embedded nuclear polyhedrosis virus (HzSNPV), with the greatest degree of inhibition occurring at low doses of viral inoculum. Additionally, the ingestion of these phytochemicals significantly prolonged the survival time of virally infectedH. zea larvae. These results suggest that the effectiveness of nuclear polyhedrosis viruses in controllingH. zea populations may be adversely affected by varieties ofL. esculentum with significant levels (eg. 3.5 mol/g wet weight) of rutin or chlorogenic acid.     

Comparative effects of two plant secondary metabolites on host-parasitoid association

Two plant-derived allelochemicals, berberine and -terthienyl (- T), were tested for their effects on the European corn borer,Ostrinia nubilalis, and its endoparasitoidDiadegma terebrans. The compounds were administered to the host insect in meridic diets, and the responses of the host larvae and parasitoids reared from treated hosts were measured in terms of growth parameters and survival. InO. nubilalis, survival to pupation and adult emergence were reduced significantly by the inclusion of berberine and -T in larval diets at a concentration of 100 g/g. However, in the parasitoid, adverse effects were much more apparent with the -T treatment than with the berberine treatment. -T and one of its metabolites were found in host larvae and in emerged adult parasitoids and their cocoons. Berberine residues were not detected. The implications of these responses to compounds of widely differing physiological properties are discussed with reference to host-plant resistance and biological control.     

Effects of Peruvian Maize Extracts on Growth, Development, and Fecundity of the European Corn Borer

Twelve Peruvian maize, Zea mays, accessions were selected because of their relatively high level of field resistance to first-generation European corn borer (ECB), Ostrinia nubilalis, larval leaf-feeding. Water extracts of freeze-dried, powdered, leaf tissue were incorporated into a standard ECB diet, fed to larvae, and the effects on larval growth, development, and fecundity were measured. Larval and pupal weights were monitored as were the time elapsed in the larval, pupal, and adult stages. Adult fecundity and egg fertility were recorded. The experiment was a randomized block design (larvae and pupae) or a completely randomized design (adults) and analyzed with ANOVA ( = 0.05). Pairwise comparisons were made between groups of insects grown on diets containing extracts from the Peruvian lines, a standard diet, or diets containing extracts of a known susceptible inbred, and a known resistant inbred line. Survival was analyzed with a chi-squared test ( = 0.05). Two Peruvian accessions significantly reduced female larval and pupal weights, extended pupal and adult development time, and decreased survival of pupae and adults. Water extracts also had a pronounced impact on males; two accessions significantly reduced pupal weight and extended the time required to pupate, and one reduced male survival to adults. The results indicate that water-soluble factors from resistant Peruvian accessions inhibit the growth, developmental time, and survival of ECB. These resistance factors could be useful in the development of maize germplasm with insect-resistant traits.     

Piperidine Alkaloids in Nitrogen Fertilized Pinus ponderosa

We fertilized individual, pole-size ponderosa pine trees at two low-quality sites and pine saplings at a relatively high-quality site, with ammonium nitrate. Six to 12 months later, we measured total %N and 2,6-disubstituted piperidine alkaloids in the foliage. The N additions raised foliar %N above deficiency levels (i.e., from 1.0–1.1% to 1.4–1.6%) at the low-quality sites, but did not elevate foliar %N in saplings at the higher quality site, where it was already (1.9%) well above critical levels. In control trees with foliar N below a threshold of 1.1%, we detected no more than trace levels of alkaloids, indicating that alkaloid production is highly constrained by N deficiency. The N additions increased mean concentrations of the predominant alkaloid, pinidine, at all three sites. Mean total alkaloid concentrations for fertilized trees at the two low-quality sites were 12 and 155 g/g dry wt higher than controls (relative increases of 12× and 4.5×, respectively). For saplings at the high-quality site, the mean total increased by 584 g/g dry wt (1.6×) with the N additions. Allocation of foliar N to alkaloids was highest in fertilized saplings (0.81%) compared to control saplings (0.53%). These findings demonstrate that foliar alkaloid concentrations can be increased by nitrogen fertilization of forest trees growing on both low- and high-quality sites. Fertilizing for the purpose of inhibiting potential herbivores may be more successful at higher quality sites where alkaloid levels are enhanced relative to food quality (foliar %N).     

Developmental inhibition ofSpodoptera litura (Fab.) larvae by a novel caffeoylquinic acid from the wild groundnut,Arachis paraguariensis (Chod et Hassl.)

A novel compound, 1-caffeoyl-4-deoxyquinic acid (1-CdQA) has been identified along with 3-caffeoylquinic acid (3-CQA) and 5-caffeoylquinic acid (5-CQA) (syn. chlorogenic acid) in the foliage ofArachis paraguariensis, a wild species of groundnut that is highly resistant to attack by the larvae ofSpodoptera litura. When neonate larvae were fed on diets treated with 3-CQA or 1-CdQA, their development was severely inhibited compared to larvae on untreated diets, and the effects were similar in nature and magnitude to those observed for larvae feeding on diets treated with 5-CQA, rutin, and quercetin. The effects of all the compounds were dose related, and their optimal concentration was approximately 3 mM, which corresponds to the total foliar concentration of both caffeoylquinic acids and quercetin diglycosides inA. paraguariensis. After 24 h, the development of third stadium larvae feeding on diets treated with 5-CQA, rutin and 1-CdQA was promoted compared to larvae on control diets, but after 96 hr larvae feeding on treated diets had gained significantly less weight than those on the control diets. 1-CdQA, 3-CQA, and quercetin dyglycosides (previously identified in the foliage ofA. paraguariensis) are considered to be valuable components in resistance of groundnuts toS. litura and may provide useful genetic markers in future breeding for resistance to this pest.     

Interactions amongHeliothis virescens larvae,cotton condensed tannin and the CryIA(c) δ-endotoxin ofBacillus thuringiensis

The potential interactions among a plant-produced allelochemical, a phytophagous insect, and an endotoxin produced byBacillus thuringiensis were investigated using purified cotton condensed tannins, the CryIA(c)-endotoxin fromB. thuringiensis subsp.kurstaki strain HD-73, and larvae ofHeliothis virescens. Purified condensed tannin from cotton fed to neonateH. virescens reduced feeding and mortality caused by insecticidal crystals ofB. thuringiensis. In fifth instars, tannin reduced relative growth rate (RGR), relative consumption rate (RCR), but antagonized the effects of the crystal-endotoxin. Tannin did not deter feeding of fifth instars in choice tests with cellulose-ester disks. Masking tannin from interacting with the dietary ingredients of artificial diets and crystal protein by encapsulation in alginate gel suggested that tannin adversely affected feeding after ingestion.These results suggest that insect control tactics that employ-endotoxins in microbial insecticides and transgenic cotton plants may not be compatible when used in conjunction with plants containing high tannin concentrations.     

The protective effect of antioxidants to a phototoxin-sensitive insect herbivore,Manduca sexta

Photo-activated plant secondary compounds have been shown to be toxic to many organisms including insects. Insect defenses include behavioral mechanisms such as light avoidance, as well as specific biochemical defenses such as antioxidants and antioxidant enzymes. These antioxidant defenses eliminate or quench the deleterious singlet oxygen and free radicals formed by these phototoxins. In this paper we examined the role of dietary antioxidants in protecting the phototoxin-sensitive insect herbivoreManduca sexta. Elevated dietary levels of the lipid-soluble antioxidants-carotene and vitamin E resulted in a concentration-dependent reduction in the mortality associated with treatment ofM. sexta larvae with the phototoxic thiophene-terthienyl. Elevated levels of dietary ascorbic acid had no effect, whereas reduced levels greatly increased the toxicity of-terthienyl. Tissue levels of antioxidants were shown to increase substantially in larvae fed antioxidant-supplemented diets. The results suggest that the ability to absorb and utilize plant-derived antioxidants could be an important defense against photo-activated plant secondary compounds and may have allowed some insects to exploit phototoxic plants.     

Fate of iridoid glycosides in different life stages of the Buckeye,Junonia coenia (Lepidoptera: Nymphalidae)

The buckeye butterfly,Junonia coenia (Lepidoptera: Nymphalidae), specializes on plants that contain iridoid glycosides. To determine the fate of these compounds in larvae, pupae, and adults of this species, we reared larvae on artificial diets with and without iridoid glycosides, and on leaves of a host plant,Plantago lanceolata (Plantaginaceae). Quantification by gas chromatography showed that newly molted third-, fourth-, and fifth-instar larvae reared on leaves ofP. lanceolata contained means of 5.13, 2.88, and 6.83% dry weight iridoid glycoside. In contrast, the mean iridoid glycoside concentration of actively feeding fifth-instar larvae was 0.28% dry weight, that of pupae was 0.19% dry weight iridoids, and adults contained no detectable iridoids. Feeding experiments suggested that this reduction in actively feeding larvae was due to the metabolism of iridoid glycosides.P. lanceolata leaves in these experiments contained a mean of 1.00% dry weight iridoid glycoside, with a 2:1 ratio of aucubin to catalpol. Calculation of iridoid consumption and utilization indices showed that larvae fed artificial diets consumed, digested, and sequestered aucubin and catalpol in similar ways. When these indices were calculated for larvae fed leaves ofP. lanceolata, catalpol was sequestered twice as efficiently as aucubin.