Isolation and identification of cotton synomones mediating searching behavior by parasitoidCampoletis sonorensis

In laboratory bioassays, the parasitoidCampoletis sonorensis was attracted to the following sesquiterpenes isolated from cotton essential oil: -humulene, -bisabolene, -caryophyllene oxide, spathulenol, -bisabolol, and a new, naturally occurring bisabolene-related alcohol, (2-p-tolyl-6-methylhept-5-en-2-ol) which we name gossonorol. This is the first report of spathulenol in cotton. -Caryophyllene, a major component of cotton, was not attractive to the parasitoids. The response of the parasitoids to these compounds and the possibility of augmenting parasitoid activity in the field by manipulating plant secondary metabolites is discussed.Synomone: A chemical produced or acquired by an organism that benefits both the emitting and receiving organisms (Nordlund and Lewis, 1976).     

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.     

Antifungal polyoxygenated fatty acids fromAeollanthus parvifolius

Two polyoxygenated fatty acids with antifungal activity have been isolated from aerial parts ofAeollanthus parvifolius (Labiatae) using bioassay-guided isolation procedures. Spectroscopic analysis was used to identify the compounds as (Z)-4,9-diacetoxy-6,7-epoxy-5-hydroxy-8-oxododeca-2-enoic acid and (Z)-4,6,7,9-tetraacetoxy-5-hydroxy-8-oxododeca-2-enoic acid. Both of these compounds inhibited spore germination ofCladosporium cucumerinum. Minimum inhibitory doses were 1 µg and 5 µg, respectively, but only the former inhibited germination ofAspergillus niger at a dose of 5 µg. Neither compound exhibited antibacterial activity. (Z)-4,9-Diacetoxy-6,7-epoxy-5-hydroxy-8-oxododeca-2-enoic acid reduced the hyphal growth ofPythium ultimum. The mode of action was not resolved but did not involve either the disruption of the cell wall membrane or the inhibition of extracellular enzymes. Application of (Z)-4,6,7,9-tetraacetoxy-5-hydroxy-8-oxododeca-2-enoic acid to seedling plants did not give any protection against fungal attack, and indeed the compound appeared to be phytotoxic.     

Factors Affecting Volatile Emissions of Intact Potato Plants, Solanum tuberosum: Variability of Quantities and Stability of Ratios

The quantity, ratios, diurnal rhythm, and correlation with foliage weight of volatile chemicals emitted by young and mature intact potato plants were determined during the fourth (morning), eighth (noon) and fourteenth (afternoon) hour of the photophase. The major compounds released by young and mature intact potato plants, Solanum tuberosum, during the photophase, were the sesquiterpene hydrocarbons -caryophyllene, (E)--farnesene, (Z,Z)--farnesene, germacrene-D, -bisabolene, and an unknown compound A. The quantity of the major volatile chemicals emitted was subject to diurnal rhythm. In young potatoes, all major compounds except unknown A showed a steady increase during the photophase, with afternoon production significantly higher than that in the morning. In mature potatoes, all compounds, including unknown A, were significantly higher during the afternoon when compared to morning. Although the variability in quantity of volatile chemicals released between individual plants was very high, the ratios were stable between the sets of replicates. The correlation of foliage weight and emission of the volatile compounds was affected by the developmental stage of the plant and the time of the day. There was a positive correlation between foliage weight and production of volatile chemicals that was strongly evident in mature potato plants, and much less evident in young plants during morning and afternoon. No positive correlation between foliage weight and production of volatile chemicals was observed during noon for either age tested. The factors affecting the volatile emissions of intact potato plants are discussed.     

Response of the Seven-spot Ladybird to an Aphid Alarm Pheromone and an Alarm Pheromone Inhibitor is Mediated by Paired Olfactory Cells

Electrophysiological responses of adult seven-spot ladybirds, Coccinella septempunctata, to (E)--farnesene, an aphid alarm pheromone, and (–)--caryophyllene, a plant-derived alarm pheromone inhibitor, were investigated by recording from single olfactory cells (neurons) on the antenna. Cells having high specificity for each of the two compounds were identified. Furthermore, these two cell types were frequently found in close proximity, with a larger amplitude consistently recorded for the cell responding specifically to (E)--farnesene. Preliminary behavioral studies in a two-way olfactometer showed that walking adults were significantly attracted to (E)--farnesene; this activity was inhibited with increasing proportions of (–)--caryophyllene. The possible ecological significance of colocation or pairing of olfactory cells for semiochemicals with different behavioral roles is discussed.     

Correspondence of soldier defense secretion mixtures with cuticular hydrocarbon phenotypes for chemotaxonomy of the termite genus Reticulitermes in North America

Soldier defense secretions from samples of Reticulitermes collected in California, Nevada, Arizona, New Mexico, and Georgia were characterized and correlated with cuticular hydrocarbon phenotypes. Twenty-seven cuticular hydrocarbon phenotypes have been defined, and soldier defense secretion (SDS) phenotypes have been described for 25 of these. Forty-five terpenoid compounds were found, including monoterpenes, sesquiterpenes, and a few diterpenes. The monoterpenes include (–)--pinene, (–)--pinene, (–)-camphene, myrcene, (Z)- and (E)-ocimene, and (–)-limonene. The major sesquiterpenes produced are (+)--cadinene, (+)--cadinene aldehyde, (–)-germacrene A, germacrene B, -himachalene, and -bisabolene. Some SDS phenotypes pair with more than one cuticular hydrocarbon phenotype; however, with two exceptions, each hydrocarbon phenotype is associated with only one SDS phenotype. These chemical characterizations lend support to the conclusion that there are numerous undescribed species of Reticulitermes in North America.     

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.     

Comparison of volatiles emitted by male caribbean and mexican fruit flies

Volatiles emitted byAnastrepha suspensa andAnastrepha ludens males were trapped and compared by GC-MS. Both species emitted previously reported nine-carbon alcohols, (Z)-3-nonenol and (Z,Z)-3,6-nonadienol, and epimeric lactones, anastrephin and epianastrephin. A third isomeric lactone, the macrolide suspensolide, which was previously known only fromA. suspensa is reported here for the first time in the volatiles ofA. ludens. We also report that both species released different proportions of the same three sesquiterpene hydrocarbons:-farnesene,-bisabolene, and-trans-bergamotene. The-bisabolene was isolated in sufficient quantity fromA. suspensa volatiles to establish its absolute configuration as the (R)-(+)-enantiomer. Some plausible biosynthetic relationships of farnesol to the isoprenoid lactones and sesquiterpenes identified in this study are discussed. Finally, we report thatA. suspensa produces the monoterpene (Z)--ocimene whileA. ludens volatiles contained limonene.     

Isolation,characterization and activity of phytotoxic compounds from quackgrass [Agropyron repens (L.)Beauv.]

Previous experiments showed that legumes grown in the presence of living or herbicidally treated quackgrass residues or extracts exhibited reduced seedling root and shoot growth and decreased nodulation and nitrogen fixation. Aqueous extracts of quackgrass shoots were most inhibitory to plant growth. Upon sequential partitioning of an aqueous extract of quackgrass shoots, the ether extract possessed the most activity and caused 50% reductions in radicle elongation of eight crop and weed species at concentrations of less than 240 g/ml (small-seeded species) and 1000 g/ml (large-seeded species). Snapbeans (Phaseolus vulgaris L. Bush Blue Lake) grown aseptically in agar containing an ether extract at 100 and 200 g/ml exhibited severe root browning, lack of root hair formation, and a two- to three-fold reduction in root and shoot dry weights. The ether extract of quackgrass shoots had no inhibitory effect on the growth of fourRhizobium species in Petri dishes or two species in broth culture. Inhibitors present in the ether extract may influence the legumeRhizobium symbiosis indirectly by reducing legume root growth and root hair formation. The ether extract of quackgrass shoots was separated using high-pressure liquid, thin-layer, and liquid column chromatography in an attempt to isolate and identify the inhibitors responsible for the inhibition of seedling growth. Two closely related flavonoid inhibitors were isolated from the ether extract. One was identified as 5,7,4-trihydroxy-3,5-dimethoxyflavone (tricin). Both flavonoids caused 50% inhibition of radicle elongation in cress (Lepidium sativum L. Burpee curly) seeds at concentrations of less than 125 g/ml. Both flavonoids were found in ether extracts of quackgrass shoots and rhizomes, but the largest amounts of both compounds occurred in quackgrass shoots collected from the field.Journal article No. 11887 of the Michigan Agricultural Experiment Station.     

Sex pheromone of aPlanotortrix excessana sibling species and reinvestigation of related species

The sex pheromone of aPlanotortrix excessana sibling species was investigated. Females were found to produce eight potential pheromone components: dodecyl acetate, tetradecyl acetate (14OAc). (Z)-5-tetradecenyl acetate (Z5-14OAc), (Z)-7-tetradecenyl acetate (Z7-14OAc), (Z)-9-tetradecenyl acetate, hexadecyl acetate, (Z)-7-hexadecenyl acetate, and (Z)-9-hexadecenyl acetate. When these compounds were bioassayed using field-trapping and wind-tunnel techniques, only 14OAc,Z5-14OAc, andZ7-14OAc were found to be behaviorally active. The sex pheromone glands of females of other species including,Planotortrix MBS,Planotortrix M,P. notophaea, Ctenopseustis servana, and aC. obliquana sibling species, were also found to containZ5-14OAc orZ7-14OAc, singly or in combination. In the case ofPlanotortrix M, the addition ofZ7-14OAc to the previously identified sex pheromone blend ofZ5-14OAc and 14OAc was found to increase trap captures of male moths of this species. Thus in these New Zealand species (and in some Australian species),Z5-14OAc andZ7-14OAc appear to be utilized in combination in pheromonal communication just as (Z)-11-tetradecenyl acetate and (E)-11-tetradecenyl acetate are used by many species of Holarctic Tortricidae in the tribe Archipini.Lepidoptera: Tortricidae: Tortricinae.     

Single and blended maize volatiles as attractants for diabroticite corn rootworm beetles

Synthetic maize volatiles and analogs dispensed singly and blended were tested for attractiveness to western (WCR, Diabrotica virgifera virgifera) and northern corn rootworm beetles (NCR, D. barberi) in maize fields. Newly identified attractants included syn-benzaldoxime, especially for NCR, and -caryophyllene for WCR females. (±)-Linalool was more effective than was (–)-linalool. Myrcene, (+)--pinene, and (–)--pinene were unattractive. Adding methyl salicylate to (±)-linalool, (+)--terpineol, or -ionone appeared to synergistically increase capture of WCR females, but dispensing the terpenes in binary blends did not. Dose–response data for methyl salicylate, (±)-linalool, and a blend of both compounds confirmed the synergy. -Caryophyllene, but not (–)--pinene, added to the latter blend produced a further synergistic increase in WCR female capture that did not vary with sesquiterpene dose from 1.0 to 100 mg. Indole addition to the same blend caused an increase in WCR female captures indicative of synergy, assuming that each did not individually lure different segments of the WCR female population. The green leaf volatiles (Z)-3-hexenyl acetate and (Z)-3-hexen-1-ol were unattractive alone and had no influence on efficacy of traps baited with 3.3 mg each of (±)-linalool, methyl salicylate, and -caryophyllene. The latter mixture captured about half as many WCR females as did 10 mg of 4-methoxycinnamaldehyde, a potent WCR attractant standard. Substituting -ionone for (±)-linalool yielded a ternary blend that captured more beetles than did the aldehyde and was unaffected by aldehyde addition. Olive oil, which has been used to sustain attractant volatilization, did not affect captures. The results show that the blending of maize volatiles has the potential to greatly improve efficacy of lures having promising applications in corn rootworm population management.     

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.     

Defensive steroids from a carrion beetle (Silpha novaboracensis)

Two novel steroids, 3,7-dihydroxy-14-pregn-4-en-15,20-dione (1) and 3,7, 20-trihydroxy-14-pregn-4-en-15-one (4), have been characterized from the rectal gland ofSilpha novaboracensis (Coleoptera: Silphidae). Judging from the function of comparable pregnanes in another species ofSilpha, the compounds may play an antipredator role.Paper No. 81 of the seriesDefense Mechanisms of Arthropods. Paper No. 80 is Eisner, T., Deyrup, M., Jacobs, R., and Meinwald, J.,J. Chem. Ecol. 12:1407–1415.     

Methodological approach to identify chemical oviposition stimulants from maize for European corn borer

The noncontingent distribution ofO. nubilalis Hbn. eggs on various corn varieties has allowed their classification as either favorable or nonfavorable hybrids. This classification is based on a chemical communication system using volatile compounds active at a distance from the insect or active when in contact with the insect as soluble sugar components. Under this hypothesis, gas chromatography is the best way of investigating for the chemical differences between corn silk extracts from the two types of hybrids. A first experiment, using a desorption-trapping system on Tenax coupled to a gas chromatograph showed a quantitative difference between two compounds from the two hybrids. A second experiment showed the advantages and disadvantages of this method and allowed the development of two complementary techniques: (1) direct extraction of corn silk with trichlorofluo-romethane and direct injection into a capillary column with an apolar stationary phase using an on-column system; and (2) extraction under vacuum of volatile compounds from corn silk and their trapping on Tenax, followed by a second desorption. This allows a direct injection on the same capillary column. The first technique allows identification by mass spectrometry of many alkanes with high molecular weights. The second technique seems to confirm the presence of phenylacetaldehyde. Both techniques show quantitative differences in the composition of corn silk extracts from favorable and nonfavorable hybrids; however, the biologically active chemicals remain unknown.     

Fertilizer requirements for specified yield targets. I. Theoretical derivation of mathematical models for the computation of soil and fertilizer nutrient efficiencies

The conventional deduction procedure of computation of soil () and fertilizer () nutrient efficiencies for the amount of fertilizer required for specified yield targets does not make provision of the amount of soil nutrient derived by crops from the available pool of soil nutrients not accounted for in the amount extracted by a soil test procedure. The derivation of two mathematical models, viz., Tamil Nadu Agricultural University Model I [TNAU Model I] and Model II [TNAU Model II] is reported in this paper which aim at computing the soil () and fertilizer () nutrient efficiencies not accounted for by the conventional method.In the case of TNAU Model I, the relationship between the nutrient uptake (U) and the soil (S) and the fertilizer (F) nutrients was established by assuming a functional relationship of the type U =S +F such that 0 1 and 0 1. In TNAU Model II the same relationship was established as U =S +F + such that 0 1, 0 1 and > 0. The term in the latter model is a measure of the amount of soil nutrient the crop absorbs from a slowly available pool of nutrients not accounted for in the amounts extracted by the soil test procedure employed or applied through fertilizer.The field verification of these models is reported elsewhere.     

Aggregation arrestant pheromone of the German cockroach,Blattella germanica (L.) (Dictyoptera: Blattellidae): Isolation and structure elucidation of blattellastanoside-A and -B

The aggregation pheromone of the German cockroach,Blattella germanica, consists of attractant and arrestant, which can be detected by olfactometer and choice-chamber assay, respectively. Both were extracted from the frass-contaminated filter paper being used as a shelter. They were separated by solvent partition withn-butanol and water. The arrestant from then-butanol phase was purified by open column chromatography and then successive HPLC isolated two major arrestant components. Spectral evidence from SI-MS, HR-EI-MS, and NMR experiments with pulse techniques provided possible structures as 1-(6-chloro-4,5-epoxy-5-stigmast-3-yl)--d-glucopyranoside and 1-(6-chloro-5-hydroxy-5-stigmast-3-yl)--d-glu-copyranoside, denoted as blattellastanoside-A and blattellastanoside-B, respectively. They represented arrestant activity as median effective doses (ED₅₀) at 0.044 (A) and 3.2 (B) nmol on 1.0 cm² of Whatman No. 1 filter paper.     

A Novel Approach for Isolation of Volatile Chemicals Released by Individual Leaves of a Plant in situ

A glass chamber designed specifically for collecting volatile chemicals from individual leaves of a plant in situ is described. The effectiveness of the chamber was demonstrated by collecting volatile chemicals from single leaves of two plant species, potato (Solanum tuberosum) and broad bean (Vicia faba), before and after mechanical damage. The glass chamber, in conjunction with thermal desorption, enables reduction of the entrainment time and thereby allows the monitoring of compounds released by leaf damage in successive 5-min periods. An intact broad bean leaf, in the middle of the day, produces small amounts of the green leaf volatiles (E)-2-hexenal and (Z)-3-hexen-1-ol. However, during the first 5 min after mechanical damage, large amounts of (Z)-3-hexenal, (E)-2-hexenal, and (Z)-3-hexen-1-ol are produced. The decline in production of (Z)-3-hexenal and (E)-2-hexenal is fast, and after 10 min, these compounds reach very low levels. (Z)-3-Hexen-1-ol shows an increase for the first 10 min and then a gradual decline. An intact potato leaf, in the middle of the day, produces very small amounts of the sesquiterpene hydrocarbons -caryophyllene and germacrene-D. After being damaged, the profile of released volatiles is different from that of broad bean. In potato, damage is associated with release of large amounts of green leaf volatiles and sesquiterpene hydrocarbons. Compounds such as (Z)-3-hexenal, (E)-2-hexenal, and (Z)-3-hexen-1-ol are released in high amounts during the first 5 min after damage, but after 10 min, these drop to very low levels. High release associated with damage is also observed for -caryophyllene, (E)--farnesene, germacrene-D, and -bisabolene. The highest level is reached 5 min after damage and 15 min later, these compounds drop to low levels. The significance of compounds released after plant damage is discussed.     

Investigation of the role of diterpenes produced by marine pulmonatesTrimusculus reticulatus andT. conica

The intertidal pulmonate limpetTrimusculus reticulatus, which is found in caves or crevices along the California coast, was previously reported to contain two novel diterpenoids, 6-isovaleroxylabda-8,13-dien-7,15-diol (1) and 2,7-diacetoxy-6-isovaleroxylabda-8,13-dien-15-ol (2). Dissection of the animals prior to extraction revealed that the diterpenoids were concentrated in the mantle, foot, and mucus, but not in the viscera. The presence ofT. reticulatus or its mucus was toxic to veliger larvae of the sabellariid reef-building tube wormPhragmatopoma californica. The major diterpenoid1 was responsible for the observed larvicidal activity. Protection against overgrowth by settling invertebrate larvae is important for the survival of this sessile filter-feeding pulmonate. A related pulmonate,T. conica, which was found in similar intertidal habitats in New Zealand, contained 6-acetoxy-7-isovaleroxylabda-8,13-dien-15-oic acid (3) and 1,7,12,18-tetraacetoxy-cholest-5-en-3-ol (4). The diterpene3 was also localized in the foot, mantle, and mucus ofT. conica but was not larvicidal toP. californica.     

Identification of a Female-Specific, Antennally Active Volatile Compound of the Currant Stem Girdler

We identified (Z)-9-octadecen-4-olide as a female-specific, antennally active compound from the currant stem girdler Janus integer Norton. Female specificity was demonstrated by gas chromatographic comparison of liquid chromatography fractions of male and female volatile emissions and whole body extracts. The -lactone was identified by coupled gas chromatographic-electroantennographic detection (GC-EAD), coupled gas chromatographic-mass spectrometric (GC-MS) analysis, microchemical reactions, and GC and MS comparison with authentic standards. GC-EAD analysis of female volatile emissions and cuticular extracts showed a single peak of activity on male antennae, which was not present in male-derived materials. Female antennae did not respond to any of the tested materials. The hydrogenation product of the natural EAD-active material was a known saturated -lactone. The mass spectrum of the dimethyl disulfide derivative of the natural -lactone was consistent with a double bond present in the 9 position. Comparison of the natural -lactone and a synthesized racemic mixture of (Z)-9-octadecen-4-olide on a chiral GC column showed the presence of a single enantiomer in the natural material.     

Soldier defense secretions of malaysian free-ranging termite of the genusLacessititermes (Isoptera,Nasutitermitinae)

Soldiers of free-ranging termites of the genusLacessititermes (Isoptera, Nasutitermitinae) secrete from their frontal glands a mixture of monoterpenes, sesquiterpenes, and diterpenes.Lacessititermes ransoneti, L. laborator, andL. species A produce species-specific secretions, the composition being most complex forL. laborator. Apart from known mono- and dihydroxytrinervitadienes, the following new diterpenes were isolated and tentatively assigned as trinervita-1(15),8(19)-dien-2,3,9,14-tetraol 2,3,14-O-triacetate, trinervita-1(15),8(19)-dien-2,3,9,14-tetraol 2,3,14-O-triacetate, 2,3,9, 14-tetraacetoxy-1 (15), 8(19)-trinervitadiene, and 2,3,11,13-tetraacetoxy-1(15),8(19)-trinervitadiene. Data on intragenus chemical variations were subjected to canonical discriminant analysis and genetic distances among the species were calculated to depict intragenus identities and affinities.