The influence of a queen-produced substance, 9HDA,on swarm clustering behavior in the honeybeeApis mellifera L.

The effect of enantiomers of the queen-produced substance, 9-hydroxy-(E)-2-decenoic acid (9HDA) on swarm clustering behavior of the honeybeeApis mellifera was studied. Caged queens were removed from the swarms at the start of each test and were replaced with small Petri dishes containing one of the following treatments: 100 gS(+) enantiomer of 9HDA, 100 gR(-) enantiomer of 9HDA, 200 g racemic (R, S) 9HDA, and a vehicle-treated control. Each swarm was considered to have dispersed when it had lost 50% of its starting weight. All treatments with 9HDA resulted in significantly longer swarm aggregation when compared with the control. Enantiomers were not shown to have different effects at theP 0.05 level of significance. However, observations on swarm behavior indicated that theR(-) enantiomer was the most active in retarding swarm dispersal.     

Sugar beet crown borer,Hulstia undulatella (Clemens) 1: Identification and field tests of female sex pheromone gland components

Electroantennogram profiles of saturated and monounsaturated 12-, 14-, and 16-carbon acetates, and 12- and 14-carbon alcohols implicated (Z)-9-tetradecen-1-ol acetate (Z9-14: Ac) as a component of the female sex pheromone ofHulstia undulatella (Clemens). Gas chromatography-mass spectrometric analysis of extract of the female sex pheromone glands showed the presence of Z9-14:Ac (8.5 ng/female), (Z)-9-tetradecen-1-ol (Z9-14:OH), and (Z)-11-hexadecen-1-ol acetate (Z11-16:Ac) in a ratio of 100421, respectively. In tests in sugar beet fields, Z9-14:Ac alone produced some trap catch. Addition of Z9-14: OH did not increase catch while addition of Z11-16:Ac eliminated catch, but addition of both Z9-14:OH and Z11-16: Ac increased catch sevenfold. A combination of Z9-14: OH and Z11-16: Ac without Z9-14: Ac did not produce trap catch. A lure of 200 g Z9-14:Ac+16 g Z9-14:OH+42 g Z11-16:Ac is suggested for use in monitoring traps.Lepidoptera: Pyralidae: Phycitinae.     

Influence of tobacco leaf surface chemicals on germination ofPeronospora tabacina adam sporangia

Chromatographic procedures were utilized to isolate and purify components of tobacco cuticular extracts and leaf surface chemicals.In vitro microbial bioassays determined the influence of these leaf surface compounds on germination and germ tube morphology ofP. tabacina sporangia, the tobacco blue mold pathogen, and to a lesser extentAlternaria alternata, the tobacco brown spot pathogen. Exposure to 10 g/cm² of - and -duvatrienemonols, sucrose esters, or hydrocarbons did not inhibit germination, whereas germination was significantly decreased bycis-abienol.cis-Abienol did not inhibit sporangial germination when combined with sucrose esters or hydrocarbons at a combined 10 g/cm². Germination of sporangia was completely inhibited by - and -duvatrienediols. In contrast to a previous report, -DVT-diol was more inhibitory than the isomer. Toxic effects of the DVT-diols were not altered by pH. Diluting the DVT-diols to less than 0.1 g/cm² resulted in a small but significant stimulation of germination. Previously, the DVT-diols had been identified only as inhibitory toP. tabacina. None of the leaf surface chemicals affected germination ofA. alternata conidia.Present Address: Department of Forestry, Biltmore Hall, North Carolina State University, P.O. Box 8002, Raleigh, North Carolina 27695-8002.     

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.     

Role of the isoflavonoid coumestrol in the constitutive antixenosic properties of “Davis” soybeans against an oligophagous insect,the mexican bean beetle

The antixenosic properties of the isoflavonoid, coumestrol, were tested in dual-choice leaf disk bioassays with the Mexican bean beetle (Epilachna varivestis Mulsant).E. varivestis preferred the methanol-r,reated (solvent-control) disk when the coumestrol concentration was 1.8 or 0.9/leaf disk. No preference was observed between the coumestrol-treated and the solvent-control disks when the coumestrol concentration was higher, at 3.6, or lower, at 0.45g/leaf disk. Coumestrol alone clearly is not responsible for the significant constitutive antixenosic properties of Davis soybeans,Glycine max (L.) Merrill, because the amount of coumestrol in these plants is significantly less than the minimum concentration which was antixenosic in this study. However, it might contribute to a constitutive antixenosis in Davis involving a profile of allelochemicals. A computer-aided densitometer, adapted to measure the leaf disk area, increased the resolution of the leaf area 250 (X)-fold as compared to the standard LI-COR leaf area meter.     

Synergism of Polygodial and trans-Cinnamic Acid on Inhibition of Root Elongation in Lettuce Seedling Growth Bioassays

A bicyclic sesquiterpene dialdehyde, polygodial did not inhibit root elongation up to a concentration of 12.5 g/ml in a lettuce seedling assay: trans-Cinnamic acid inhibited the elongation by 50% at 1.2 g/ml (8.1 M). The inhibitory activity of trans-cinnamic acid was enhanced 17-fold when used in combination with 6.25 g/ml (26.5 M) of polygodial. A decrease in the pH of the medium was observed during normal seedling growth, indicating transport of protons from the cells by a plasma membrane H⁺-ATPase. The inhibitory effect of trans-cinnamic acid on the elongation was reduced to some extent in 2 mM phosphate buffer (pH 7.0) during seedling growth. Although polygodial did not inhibit the activity of H⁺-ATPase in the plasma membrane fraction of roots in normally growing seedlings, a decrease in activity was found in the fraction obtained from seedlings incubated with 20 g/ml of polygodial. These results suggest that polygodial functions synergistically with trans-cinnamic acid in the inhibition of root elongation via restriction of proton transport from the cytoplasm of germinated cells.     

Attraction of 3-Methyl-1-Butanol and Ammonia Identified from Enterobacter agglomerans to Anastrepha suspensa

Tests demonstrated that volatile chemicals emitted from Enterobacter agglomerans, a bacterium that has been isolated from adults as well as fruit infested with larvae of the Caribbean fruit fly, Anastrepha suspensa (Loew) and other pest fruit flies, are attractive to female A. suspensa in laboratory bioassays. 3-Methyl-1-butanol and ammonia were identified as the two primary volatile chemicals released from active cultures of E. agglomerans. No 3-methyl-1-butanol and little ammonia (16.0 g/hr) are released from sterile tryptic soy agar plates. E. agglomerans-inoculated tryptic soy agar plates, however, released an average of 1.5 ± 0.53 g/hr 3-methyl-1-butanol and 332.9 ± 239.16 g/hr ammonia after 24 hr of growth. 3-Methyl-1-butanol lures were formulated in a membrane-based system to provide a constant release rate of synthetic chemical. Release rates ranged from 0.046 ± 0.007 to 12.16 ± 2.76 g/hr. In laboratory tests, equal numbers of females were captured in response to ammonium carbonate lures that released ammonia at the rate of 100 g/hr and to 3-methyl-1-butanol lures that released 12.16 ± 2.756 g/hr of synthetic material. The combination of the two lures was more attractive than ammonia alone. Availability of lures formulated for a range of 3-methyl-1-butanol release rates will facilitate field tests of this putative microbial attractant and may lead to a better understanding of the role of bacteria in the ecology of pest fruit flies.     

Identification of triterpenoid feeding deterrent of red pumpkin beetles (Aulacophora foveicollis) fromMomordica charantia

The triterpenoids isolated from the leaves ofMomordica charantia Linn (bitter gourd) were found to elicit feeding-deterrent activity against red pumpkin beetles (Aulacophora foveicollis Lucas). The most abundant triterpenoid which deterred feeding was identified as momordicine II, 23-O--glucopyranoside of 3,7,23-trihydroxycucurbita-5,24-dien-19-al. A concentration of 3200g/ml and above of the triterpenoids caused significant reduction of feeding by red pumpkin beetles in in vitro bioassay experiments, which compared favorably with the levels of triterpenoids inM. charantia leaves found in nature.     

Evaluation of vetiver oil and seven insect-active essential oils against the Formosan subterranean termite

Repellency and toxicity of 8 essential oils (vetiver grass, cassia leaf, clove bud, cedarwood, Eucalyptus globules, Eucalyptus citrodora, lemongrass and geranium) were evaluated against the Formosan subterranean termite, Coptotermes formosanus Shiraki. Vetiver oil proved the most effective repellent because of its long-lasting activity. Clove bud was the most toxic, killing 100% of termites in 2 days at 50 g/cm². The tunneling response of termites to vetiver oil also was examined. Vetiver oil decreased termite tunneling activity at concentrations as low as 5 g/g sand. Tunneling and paper consumption were not observed when vetiver oil concentrations were higher than 25 g/g sand. Bioactivity of the 8 oils against termites and chemical volatility were inversely associated. Listed in decreasing order of volatility, the major constituents of the 8 oils were: eucalyptol, citronellal, citral, citronellol, cinnamaldehyde, eugenol, thujopsene, and both - and - vetivone. Vetiver oil is a promising novel termiticide with reduced environmental impact for use against subterranean termites.     

Defensive secretions of three species ofAcilius (Coleoptera: Dytiscidae) and their seasonal variations as determined by high-pressure liquid chromatography

A rapid, sensitive, stable, and quantitative high-pressure liquid chromatographic technique was developed for the analysis of defensive secretions obtained from the pygidial and prothoracic glands of dytiscids. Methods were developed for both normal phase (Porasil) and reverse phase (Bondapak C₁₈) columns. The applicability of this technique was demonstrated when defensive compounds ofAcilius semiculcatus, A. sylvanus andA. mediatus were isolated, identified, and quantitated. No major differences were found in the composition of the defensive secretions between the three species. The seasonal defensive titer ofA. semisulcatus was determined from June through October 1977. The pygidial defensive agents (benzoic acid,p-hydroxybenzaldehyde, and methylp-hydroxybenzoate varied from 9.0 to 67.8 g and exhibited a maximum in July, and a steroid from the prothoracic gland varied from 7.1 to 33.2 g and was maximum in October.     

Quantitative and qualitative variation in male pheromones ofPhragmatobia fuliginosa andPyrrharctia isabella (Lepidoptera: Arctiidae)

The dihydropyrrolizine pheromones, hydroxydanaidal and danaidal, were identified from the scent organs of malePhragmatobia fuliginosa (L.) andPyrrharctia isabella (J.E. Smith). Qualitative and quantitative GLC analyses were conducted on ca. 80 field-collected males of each species. The total pheromone titer was distributed bimodally in each species with most males having either a small amount (< 10 ng) of pheromone or a large amount (1–10 g inPyrrharctia and 0.3–3 g inPhragmatobia).Pyrrharctia males in the 1- to 10-g range had a predominance of hydroxydanaidal, with little if any danaidal. MostPhragmatobia males in the 0.3- to 3-g range had danaidal with little if any hydroxydanaidal. These compounds elicited a courtship response in sexually receptive females of both species. A bioassay based on this response was used to measure the thresholds of female response to these compounds.Pyrrharctia females were more sensitive to (R)-(–)-hydroxydanaidal than to danaidal.Phragmatobia females were more sensitive to danaidal then to (R)-(–)-hydroxydanaidal.     

Caste-dependent pheromones in the head of the antTetramorium caespitum

The heads of maleT. caespitum contain 4-methyl-3-hexanol (1.7 g, only the erythro isomer(s) detected) and 4-methyl-3-hexanone (0.8 g). The heads of alate females contain 0.1 g of each compound, whereas only the alcohol (0.1 g) was found in the heads of workers. Both compounds act as attractants for the workers and their possible functions are discussed.'Chargé de Recherche du F.N.R.S.Chercheur qualifié du F.N.R.S.     

Leaf surface chemicals fromNicotiana affecting germination ofPeronospora tabacina (adam) sporangia

A bioassay was used to evaluate the effects of cuticular leaf components, isolated fromN. tabacum, N. glutinosa (accessions 24 and 24a), and 23other Nicotiana species, on germinationof P. tabacina (blue mold). The leaf surface compounds included- and-4,8,13,-duvatriene-l,3-diols (DVT-diols), (13-E)-labda-13-ene-8-,15-diol (labdenediol), (12-Z)-labda-12,14-diene-8-ol (cis-abienol), (13-R)-labda-8,14-diene-13-ol (manool), 2-hydroxymanool, a mixture of (13-R)-labda-14-ene-8,13-diol (sclareol), and (13-S)-labda-14-ene-8,13-diol (episclareol), and various glucose and/or sucrose ester isolates. The above in acetone were applied onto leaf disks of the blue moldsusceptibleN. tabacum cv. TI 1406, which was then inoculated with blue mold sporangia. Estimated IC₅₀ values (inhibitory concentration) were 3.0g/cm² for-DVT-diol, 2.9/cm² for-DVT-diol, 0.4g/cm² for labdenediol and 4.7g/cm² for the sclareol mixture. Manool, 2-hydroxymanool, andcis-abienol at application rates up to 30g/cm² had little or no effect on sporangium germination. Glucose and/or sucrose ester isolates from the cuticular leaf extracts of 23Nicotiana species and three different fractions fromN. bigelovii were also evaluated for antimicrobial activity at a concentration of 30g/cm². Germination was inhibited by >20% when exposed to sugar esters isolated fromN. acuminata, N. benthamiana, N. attenuata, N. clevelandii, andN. miersii, and accessions 10 and 12 ofN. bigelovii. These results imply that a number of compounds may influence resistance to blue mold in tobacco.     

Variation in aphid alarm pheromone content among glandular and eglandular-hairedMedicago accessions

Pea (Acyrthosiphon pisum Harris) and blue alfalfa aphid (A. kondoi Shinji) deterrency in alfalfa (Medicago saliva L.) may result from incorporating higher levels of the aphid alarm pheromone,(E)--farnesene relative to(E)--caryophyllene. We evaluated five eglandular and two glandular-haired alfalfa accessions for differences in(E)--farnesene and(E)--caryophyllene content under glasshouse conditions using supercritical fluid extraction and gas chromatography. In addition, pea and blue alfalfa aphid olfactory behavioral tests were conducted uponMedicago species containing different ratios of(E)--famesene relative to(E)--caryophyllene. No differences in(E)--caryophyllene content were observed among the seven entries (=0.42 ng/g plant material). Significant differences (P 0.05) among entries were observed for(E)--famesene content, with KS94GH6 exhibiting the highest (1.18 ng/g), and CUF 101 the lowest levels (0.35 ng/g). Elite tetraploid sources possessed significantly lower levels (=0.42 ng/g) of(E)--farnesene than did wild and cultivated diploid accessions (=0.83 ng/g). Olfactory behavioral tests for both the pea and blue alfalfa aphids demonstrated KS94GH6 repelled aphids while cultivated alfalfa types attracted aphids in each case. Previously demonstrated aphid resistance in diploid KS94GH6 may result from superior(E)-- farnesene levels, but(E)--farnesene is probably not a factor in cultivated alfalfa resistance. Finally, accession KS94GH6 could act as an excellent germplasm resource for the incorporation of higher(E)--farnesene levels into cultivated alfalfa.Research supported by the New Mexico State University Agricultural Experiment Station.     

Interactions between pheromone traps with different strength lures for the pine beauty moth,Panolis flammea (Lepidoptera: Noctuidae)

Catches of malePanolis flammea in traps baited with lures containing 25 g of sex attractant are enhanced when in the proximity of traps baited with 125-g lures. The degree of enhancement is increased as the intertrap distance is decreased, and when the low-dose trap is upwind of the high-dose one. The patterns of alteration in trap catch suggest that moths initially attracted by one lure may divert into other traps, which may be either upwind of the original trap (overshooting), or downwind (undershooting). Overshooting can result in up to fivefold increases in catch and may provide a useful method for estimating the attractant zone of particular lure/trap combinations.     

Onion fly trap catch as affected by release rates ofn-dipropyl disulfide from polyethylene enclosures

n-Dipropyl disulfide loadings of at least 1 l and 10 l in polyethylene enclosures were required to effect significant trap catches of onion fly,Hylemya antiqua (Meigen), females and males, respectively. Corresponding release rates for this threshold for trap catch were ca. 100 g/hr. Catches increased with loadings up to 100 l/capsule but then plateaued. As determined gravimetrically, the release rate from enclosures containing a reservoir of chemical remained constant under isothermal conditions but increased exponentially as a function of temperature.Diptera: AnthomyiidaePaper No. 9422 of the Michigan State University Agricultural Experiment Station. Received for publication April 29, 1980.     

Linamarin and histamine in the defense of adultZygaena filipendulae

We determined the protective values of histamine and linamarin to an aposematic moth,Zygaena filipendulae. Using ion-exchange resin techniques, we found that the mean histamine concentrations in the wings were 0.061 ± 0.047 g/mg and 0.013 ± 0.0051 in the moths' bodies (totals: 0.586 g and 2.921 g, respectively, all wet weights). Average HCN evolution (mainly from the bitter cyanogen linamarin) from the wings was 0.049 ± 0.41 g/mg (0.426 g/ml of linamarin could produce this amount of HCN) and 0.029 ± 0.0026 g/mg HCN (0.281 g/mg linamarin) evolved from the bodies (total linamarin 4.09 g and 61.258 g, respectively, all wet weights). Therefore, higher concentrations of toxicants were found in the part of the body most liable to initial attack. We found, in offering various toxic solutions to 10 common quails, that 0.1% linamarin (mean linamarin consumed equal to about 70% of the average total wing content) but 1.0% histamine (mean histamine consumed equal to that found in about 8.9 average wing sets) solutions significantly lowered drinking rates. However, combination solutions were still effectively aversive at 0.001% histamine plus 0.028% linamarin. This synergism would allow a moth under local abiotic or dietary stress to elaborate substantially less of one or both compounds than that normally synthesized. The implications to kin selection are discussed.     

Effects of juglone on growth,photosynthesis, and respiration

The impacts of juglone on plant growth and several other physiological functions were evaluated in this study. Juglone inhibitedLemna minor growth, chlorophyll content, and net photosynthesis at treatments between 10 and 40M. Soybean leaf disks vacuum infiltrated with as little as 10M juglone had reduced photosynthesis. Oxygen evolution by chloroplasts isolated fromPisum sativum was inhibited by juglone with an I₅₀ of 2M. Micromolar treatments of juglone stimulated oxygen uptake in mitochondria isolated fromGlycine max. These data suggest perturbations of chloroplast and mitochondrial functions may contribute to plant growth reductions observed in juglone-mediated allelopathy.This study was supported by National Science Foundation grant STI-8902066.     

Aggregation Pheromone of the Cereal Leaf Beetle: Field Evaluation and Emission from Males in the Laboratory

The previously identified, male-specific compound of the cereal leaf beetle (CLB, Chrysomelidae; Oulema melanopus), (E)-8-hydroxy-6-methyl-6-octen-3-one, was studied further with respect to field activity and emission rate from male beetles. In a 5-week field experiment in Oregon, the compound was shown to function as an aggregation pheromone in attracting male and female CLBs migrating from overwintering sites in spring. Traps baited with the synthetic compound (500 g per rubber septum) caught 3.3 times more CLBs than control traps. Lower doses of the pheromone (50 and 150 g) were less attractive than the 500 g dose. One relatively abundant, volatile compound from the host plant (oats), (Z)-3-hexenyl acetate, that elicited responses from beetle antennae was not attractive, either by itself or as a synergist of the pheromone. Both sexes were captured about equally for all treatments. We also measured daily pheromone emission by male beetles in the laboratory. Individual males feeding on oat seedlings under greenhouse conditions emitted as much as 6 g per day, which is about 500 times higher than had been previously observed under incubator conditions. The pheromone emission rate was at least five times higher during the day than at night, and in one male, emission spanned a period of 28 d. The release rate of synthetic pheromone from the 500 g septa was very similar to the maximum from single males; thus, future experiments should evaluate even higher doses. The field results indicate that the pheromone has potential as a monitoring tool for early detection of CLBs as they move from their overwintering sites into newly planted cereal crops in spring.     

Fate of quinolizidine alkaloids through three trophic levels:Laburnum anagyroides (Leguminosae) and associated organisms

The quinolizidine alkaloids (QA) of golden rain,Laburnum anagyroides, and those of phytophagous insects associated with the plant, as well as of parasitoids of the latter, were analyzed by capillary GLC and GLC-MS. The alkaloid content in samples of vegetative plant parts was high at the beginning of the season, then decreased, while that of reproductive organs was high throughout flowering, pod formation, and maturation. The analyses showed that the QA of the plant passed through two higher trophic levels (herbivorous insects and their parasitoids) and that the alkaloid pattern changed little during the passage. The alkaloids were present in two phytophagous insect species associated with golden rain: the predispersal seed predator,Bruchidius villosus [5–13g/g fresh weight (fw)], andAphis cytisorum (182–1012g/g fw), an aphid that feeds on shoots, leaves, and inflorescences. Braconid and chalcidoid parasitoids emerging from the bruchid host also contained alkaloids (1.3–3g/g fw), as did three foraging ant species,Lasius niger, Formica rufibarbis, andF. cunicularia (45g/g fw), that visited the aphid colonies or honeydew-covered leaves of aphid-infested plants. The hypothesis that developing bruchid larvae and/or the plant manipulate QA supply to infested seeds was not supported, because QA content of leftover endosperm in seeds after bruchid development was similar to that of uninfested seeds. The frass of developing bruchid larvae was rich in QA (31g/ g dry weight). While aphids sequestered, the bruchid larvae took up and eliminated QA with the frass without chemical transformation.