Isolation,identification, and synthesis of a female sex pheromone of the navel orangeworm,Amyelois transitella (Lepidoptera: Pyralidae)

A sex pheromone of the navel orangeworm,Amyelois transitella (Walker), was obtained from ether rinses of the sex pheromone gland of calling females. The pheromone was isolated by means of liquid and gas chromatography and was identified as one of four possible geometrical isomers of 11,13-hexadecadienal by means of spectroscopic and microchemical methods. Synthesis and laboratory bioassay of all four isomers revealed that only the (Z,Z) isomer was biologically active. (Z,Z)-11,13-hexadecadienal elicited quantitatively similar activation and attraction responses byA. transitella males as did the natural product.Mention of a commercial or proprietary product in this paper does not constitute an endorsement of that product by the USDA. Accepted for publication February 28, 1979.     

Structure-activity relationship of flavones as growth inhibitors of the navel orangeworm

A series of flavones with widely varying degrees of substitution was fed to neonate larvae of the navel orangeworm. Growth of navel orangeworm larvae is inhibited by 5-methoxy flavones and flavone itself; 5-hydroxy flavones do not inhibit growth. Host resistance of citrus fruit to attack by the navel orangeworm might be due to the 5-methoxy flavones that occur in the peels.     

Stereospecific synthesis of (Z,Z)-11,13-hexadecadienal,a female sex pheromone of the navel orangeworm,Amyelois transitella (Lepidoptera: Pyralidae)

A synthesis of (Z,Z)-11,13-hexadecadienal, a compound previously identified as the major component of the sex pheromone of the female navel orangeworm,Amyelois transitella (Walker), is described. The key step is the reduction of an unsymmetrical conjugated diyne with dicyclohexylborane. This reduction produced theZ,Z isomer with less than 0.5% total of other isomers.Mention of a commercial or proprietary product in this paper does not constitute endorsement of that product by the USDA.     

Structure-activity relationship of unsaturated fatty acids as mosquito ovipositional repellents

Various straight-chain unsaturated fatty acids from C₁₄ to C₂₄ were evaluated for their ovipositional repellency against gravid females of the southern house mosquitoCulex quinquefasciatus Say, and the relationship between the structures of the fatty acids and their ovipositional repellency was determined. A double bond withZ configuration was prerequisite for an unsaturated fatty acid to be highly repellent;E isomers were less active or even inactive. No relationship was found between the repellency and the number of double bonds in the unsaturated fatty acids. In C₁₈ monounsaturated fatty acids, (Z)-9 acid was more active than (Z)-11 and (Z)-6 acids, indicating that a double bond at the 9 position rendered an acid highly repellent. Among (Z)-9-alkenoic acids of different chain lengths, the most repellent was C₁₈ acid which was also more active than (Z)-11-C₂₀, (Z)-13-C₂₂, and (Z)-15-C₂₄ acids. Oleic[(Z)-9-octadecenoic]acid, which met all these criteria, was the most ovipositionally repellent among the unsaturated fatty acids tested.Diptera: Culicidae.     

Identification of olfactory cues used in host-plant finding by diamondback moth,Plutella xylostella (Lepidoptera: Plutellidae)

Olfactory attraction of female diamondback moths (Plutella xylostella) to odors of intact and homogenized host plants, as well as individual compounds characteristic of host plants, were investigated by behavioral and electrophysiological methods. Moths were attracted to odors ofBrassica juncea andB. napus seedlings in a Y-tube bioassay. Solvent fractions of homogenizedB. juncea leaves were attractive to moths whether or not isothiocyanates (IC) were present. Moths were attracted in Y-tube bioassays and to field traps baited with individual ICs. Volatiles fromB. juncea andB. napus elicited an electroantennogram (EAG) response and were attractive in the Y-tube bioassay. Allyl IC was shown to be the attractive component in homogenized plant volatiles but was found to be virtually absent from intact plant volatiles. Gas chromatographic fractionation of intact plant volatiles revealed a terpene-containing fraction to be most attractive to the moths. We were unable to isolate individual attractive compounds from this fraction. Our results suggest that certain elements of this fraction, possibly in combination, are important olfactory cues for host-plant finding by the diamondback moth with mustard oils playing an important and possibly synergistic role, particularly when plants are damaged.     

Feeding preferences of spruce budworm (Choristoneura fumiferana Clem.) larvae to some host-plant chemicals

Single-choice and three-choice tests were used to determine the relative importance of host-plant chemical extracts in eliciting feeding by spruce bud worm larvae. Water-soluble components of the host trees are the most important and, of these, the sugar and glycoside fractions are the most stimulating. The amino acid and organic base fractions have no apparent effect. The organic acid fractions deter feeding slightly.Lepidoptera: Tortricidae.Supported by the National Sciences and Engineering Research Council of Canada, Operating Grant No. A9723 to P.J.A.     

Chemically mediated ovipositional behaviors of the european corn borer,Ostrinia nubilalis (Lepidoptera: Pyralidae)

The volatile plant sesquiterpenoids farnesene, nerolidol, and farnesol were tested to determine their effect on European corn borer,Ostrinia nubilalis, oviposition during the first six nights of the adult stage. Adult European corn borers were released into cages designed to encourage oviposition on eight glass plates randomly arranged on top of the cage: four coated with either 80µg/cm² farnesene, nerolidol, or farnesol, and four coated with the solvent methylene chloride. The setup was used as a binary choice test. Farnesene was preferred by females because they deposited significantly more egg masses on plates coated with the compound. Nerolidol had no effect. Farnesol, by contrast, deterred oviposition: the number of egg masses was significantly lower on plates covered with that compound. These data show that structural modification at a single terminal functional group in these compounds affects the ovipositional behavior of European corn borer females.This paper reports the results of research only. Mention of a proprietary product does not constitute an endorsement or a recommendation for its use by USDA.     

Sesamia calamistis calling behavior and its role in host finding of egg parasitoids Telenomus busseolae,telenomus isis,and Lathromeris ovicida

In the Republic of Benin, the African pink stem borer Sesamia calamistis Hampson is kept under control by the two scelionid egg parasitoids Telenomus busseolae (Gahan) and T. isis (Polaszek). A third species, the trichogrammatid Lathromeris ovicida Risbec, although commonly obtained from egg batches, is of low importance. High egg and egg batch parasitism by Telenomus spp. at low host densities suggests excellent host-finding capacity. Previously it was hypothesized that pheromones produced by calling S. calamistis virgins play an important role in attracting the parasitoids to the area where oviposition is likely to occur. Thus, after determining the effect of age (1–4 days) on the calling rhythm of S. calamistis, 1-day-old virgin females were chosen to investigate the response of the parasitoids to calling and noncalling females and two empty controls, in a four-arm olfactometer. The results showed that calling started earliest and lasted longest with 1-day-old females, but was similar for 2- to 4-days-old females. The percentage of females calling was higher in early than latter scotophases. The highest percentage of active females was obtained between 8 and 9 hr after the beginning of the first scotophase. All three parasitoid species responded to calling females, while there were no differences between noncalling females and the empty controls. The response pattern was similar for the two Telenomus spp. but considerably weaker for L. ovicida, reflecting differing host specificity of the three parasitoid species.     

Monooxygenase system of <Emphasis Type="Italic">Bacillus megaterium</Emphasis> ALA2: Studies on palmitic acid oxidation products

We identified many novel oxygenated FA produced from linoleic acid by microbial strain ALA2: 12,13,17-trihydroxy-9(Z)-octadecenoic acid (12,13,17-THOA); 12,13,16-trihydroxy-9(Z)-octadecenoic acid (12,13,16-THOA); 12-hydroxy-13,16-epoxy-9(Z)-octadecenoic acid; and 12,17;13,17-diepoxy-16-hydroxy-9(Z)-octadecenoic acid. 12,13,17-THOA, the main product, inhibits the growth of some plant pathogenic fungi. Recently, we reclassified strain ALA2 as Bacillus megaterium ALA2 NRRL B-21660 and opened a possible link with the well-studied catalytically self-sufficient P450 monooxygenase of Bacillus megaterium ATCC 14581 (NRRL B-3712) and B. subtilis strain 168 (NRRL B-4219). Now we have found that strain ALA2 also oxidizes palmitic acid into three oxygenated products: 13-, 14-, and 15-hydroxy palmitic acids. This indicates that strain ALA2 also possesses a monooxygenase system similar to the abovementioned well-known strains. These data facilitate studies on the oxygenase system of strain ALA2.     

Oviposition response ofLobesia botrana females to long-chain free fatty acids and esters from its eggs

Avoidance of occupied ovisposition sites supposes that females perceive information related to their own progency. Fatty acids identified from egg extracts have been reevaluated using a different extraction method, and we have investigated the dose-dependent oviposition response of European grape vine moths (Lobesia botrana) to myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, methyl palmitate, methyl oleate, and ethyl palmitate; all except ethyl palmitate have been identified from eggs ofL. botrana. A methylene dichloride extract of eggs fromL. botrana revealed the presence of saturated free fatty acids (myristic, palmitic, and stearic) and unsaturated acids (palmitoleic, oleic, linoleic, and linolenic) in amounts ranging from 3.9 ng/egg equivalent for myristic acid to 30 ng/egg equivalent for palmitic and oleic acids. The extract also contained traces of methyl palmitate and methyl stearate. The greatest avoidance indexes were observed in response to palmitic, palmitoleic, and oleic acids. All the other compounds tested caused weaker responses. A reduction in the number of eggs laid was observed when moths were exposed to each of the esters applied at 0.3 µg per application spot. Reduction in eggs laid was also observed at a 10-fold higher dose of oleic acid. The present results confirm that general and simple molecules can be involved in the regulation of oviposition site selection and that they may participate in chemical marking of the eggs.     

Identification of host-plant chemicals stimulating oviposition by swallowtail butterfly,Papilio protenor

The ovipositional response of a Rutaceae-feeding papilionid butterfly,Papilio protenor, toCitrus host plants was evoked by the synergistic action ofl-(–)-stachydrine,d-(–)-quinic acid, (–)-synephrine, andl-(–)-proline that characterize the chemical compositions of the leaves and epicarp ofCitrus plants (C. natsudaidai andC. unshiu). The stimulatory activity of their mixture was enhanced by the addition of flavanone glycosides, naringin and hesperidin, which coexist in these plants and have previously been demonstrated to serve as oviposition stimulants. However, sugars such as sucrose, glucose, and inositols, which abound in plant tissues, exerted no effect on egg-laying by the females. On the other hand, chlorogenic acid present in the leaves of another host plant,Fagara ailanthoides, was found to act as an excellent synergist. However, there existed significant qualitative dissimilarities between the chemical compositions of the leaves ofC. unshiu andF. ailanthoides. This strongly suggests thatP. protenor is likely to utilize different categories of compounds as chemical cues in recognizing each plant as a host.     

Chemistry of a Mating Plug in Bumblebees

In the bumblebee B. terrestris males transfer a mating plug into the queen's sexual tract shortly after sperm transfer. The plug is a sticky, opaque secretion of the male accessory gland. In order to clarify the meaning of the mating plug, we collected the plug substance directly from the male's accessory gland and identified the chemical substances present with gas chromatography. The main compounds found in the mating plug were four fatty acids (palmitic, linoleic, oleic, and stearic acids) and a cyclic peptide (cycloprolylproline). Mixing the four fatty acids resulted in a similar sticky, opaque mass as found in natural plugs, indicating that cycloprolylproline is not necessary for the physical attributes of the plug. The function of the fatty acids may therefore be to build up a physical barrier, optimizing sperm placement before the spermathecal duct or preventing sperm backflow. Cycloprolylproline, on the other hand, may influence female mating behavior so as to reduce her receptivity. In fact, peptides are known to reduce female receptivity in other insects. This would explain why queens of B. terrestris are only singly mated, although multiple mating is beneficial during the colony cycle with respect to parasitism and fitness.     

Identification of (e)-11-hydroxy-9-octadecenoic acid and (E)-9-hydroxy-10-octadecenoic acid by biotransformation of oleic acid by Pseudomonas sp. 32T3

Pseudomonas sp. 32T3, a newly identified strain originally isolated from a vegetable oil-contaminated soil, produces three monohydroxy acids—(E)-11-hydroxy-9-octadecenoic acid, (E)-10-hydroxy-8-octadecenoic acid, and (E)-9-hydroxy-10-octadecenoic acid—as bioconversion products of oleic acid. The bacterial cells were grown in a mineral medium containing oleic acid as the main carbon substrate. The compounds were identified as the corresponding methyl esters on the basis of their chromatographic and spectroscopic (¹H and ¹³C nuclear magnetic resonance and gas chromatography-mass spectrometry) features.     

Influence of different habitats and mating on olfactory behavior of onion flies seeking ovipositional hosts

Using traps baited with natural and synthetic onion volatiles, we examined the effects of different habitats and mating on the olfactory behavior of laboratory-reared and wild onion flies. Rankings of olfactory treatments as host-finding stimuli for females were dependent on their mating status and the habitat in which they were foraging. In habitats devoid of hosts, traps baited with individual alkyl sulfides were as effective as 4-day-old chopped onions and more effective than 1-day-old onions in eliciting host-finding behavior in laboratory-reared unmated females (LUF) and laboratory-reared mated females (LMF). However, upwind dispersal and percent recapture were always significantly greater in LUF. In one experiment, Pr₂S₂ was 19 times more attractive to LMF in a fallow field, as than it was in an onion field. Reduced effectiveness of alkyl sulfides as host-finding stimuli in onion fields probably results in part because they are less findable, but more importantly because of a change in searching behavior after females have mated. Evidence to support the latter contention is that traps baited with alkyl sulfides and onions were equally findable by unmated females in both habitats. The behavior of LMF was identical to that of wild females, whereas the behavior of LUF was identical to wild males. The hypothesis that olfactory host-finding behavior in onion flies is modified by the resource level was upheld. Alkyl sulfides appear to be the primary, and possibly the only, chemical effectors of host-finding at the patch level of resource distribution, whereas the complex blend emitted by aged, chopped, or damaged onions appears to be acting at the final level of host-finding, while egg-laying females are moving between adjacent hosts in search of an optimal oviposition site.     

Oil content and fatty acid composition of seed oil from guayule plants with different chromosome numbers

Guayule, a perennial desert plant, is being developed for domestic production of natural rubber, a strategic commodity for which the United States presently depends totally on foreign sources. At present, rubber alone is not sufficient to make guayule a commercial crop, and additional revenues are being sought from by-products. Because guayule flowers profusely during several years of growth before it is harvested for rubber, seed may also contribute to the economics of guayule production. Seed from 120 plants, including 20 genotypes with 36, 37, 54 and 72 chromosomes, were analyzed for oil content and fatty acid composition. Oil content ranged from 17.1 to 30.5%. On average, seed from diploid and aneuploid plants (with 36 and 37 chromosomes) contained 40.4% more oil than the seed from polyploid plants. The oil consisted of four fatty acids—palmitic (8.7–11.5%), stearic (3.7–6.2%), oleic (6.5–13.9%) and linoleic (69.1–80.2%)—at all ploidy levels. Guayule seed oil was similar to the seed oil from high-linoleic safflower varieties. The use of genetic variation to increase seed yield and seed oil will depend on the absence of negative correlation between oil and rubber production.     

Female sex pheromone of the melonworm,Diaphania hyalinata (Lepidoptera: Pyralidae), and analysis of male responses to pheromone in a flight tunnel

Ten C₁₆ chain-length compounds were identified from heptane extracts of ovipositors of female melonworm,Diaphania hyalinata (L.). The major constituents of the extracts were (E)-11-hexadecenal and (E,E)-10,12-hexadecadienal [(E,E)-10,12–16:Ald] and the alcohols and acetates of these olefins were found in trace amounts (<2%). Extracts also contained traces of (E,Z)- and (Z,Z)-10,12-16:Ald, hexadecanal, and 1-hexadecanol. Analysis of the behavioral responses of males to synthetic mixtures of these compounds and responses to ovipositor extracts in a flight tunnel showed that a synthetic mixture of the 10 compounds elicited a behavioral repertoire from males that was indistinguishable from that elicited by ovipositor extract. Flight tunnel studies also indicated that six of the 10 compounds probably represent the essential components of the female's sex pheromone.Supported in part by USDA Cooperative Agreement No. 58-32U4-1-299. Scientific Article No. A-4073, Contribution No. 7058 of the Maryland Agricultural Experiment Station. Mention of a commercial product in this paper does not constitute a recommendation by the U.S. Department of Agriculture.     

Characterization of monomers produced from thermal high-pressure conversion of meadowfoam and oleic acids into estolides

The monomers produced from thermal high-pressure conversion of meadowfoam or oleic acids into estolides were characterized as a complex mixture of fatty acids. Mild reaction conditions produced little change in the starting acids. However, vigorous reaction conditions,e.g. ≥3 h at 250°C with stirring, significantly altered the starting fatty acids.Cis/trans isomerization occurred readily, with the proportion oftrans isomers reaching 57%. In addition, the double bonds migrated throughout all positions of the hydrocarbon chain with concentrations diminishing outward from the starting double bond position. Branching was also observed to a small extent under these conditions and was even more pronounced in the absence of water. Lactones were also identified in the reaction mixture, with contents near 16% in the meadowfoam series. All products can be explainedvia carbocation rearrangement mechanisms that result from protonation of the starting olefins.     

Chemically mediated host finding byBiosteres (Opius) longicaudatus,a parasitoid of tephritid fruit fly larvae

Host finding by the fruit fly parasitoidBiosteres (Opius)longicaudatus Ashmead was found to involve attraction to fermentation products emanating from rotting fruit, a probable site for location of host larvae. Bioassays conducted in the greenhouse with all saturated 1-, 2-, and 3-carbon primary alcohols, aldehydes, and organic acids indicated that acetaldehyde was the most active agent followed by ethanol and acetic acid. Rotting fruit was attractive irrespective of the presence of host larvae, and attraction was attributed to fungal fermentation products rather than to kairomones from host larvae.     

Positional and geometrical isomers of linoleic acid in partially hydrogenated oils

The geometrical and positional isomers of linoleic acid of a partially hydrogenated canola oil-based spread were isolated and identified. Through partial hydrazine reduction and mass spectral studies,cis-9,trans-13 octadecadienoic acid was identified as the major isomer. Other quantitatively important isomers characterized werecis-9,trans-12;trans-9,cis-12 andcis-9,cis-15. These four were also the major isomers in margarine based on common vegetable oils. A number of minor isomers were detected and some structures identified weretrans-9,trans-12;trans-8,cis-12;trans-8,cis-13;cis-8,cis-13;trans-9,cis-15;trans-10,cis-15 andcis-9,cis-13. The proportions of the various isomers are given for some margarines in the Canadian retail market. The amounts oftrans-9,trans-12 isomer in Canadian margarines were generally below 0.5% of the total fatty acids.     

Influence of Some Fatty Acids on Oviposition by the Bruchid Beetle, Callosobruchus maculatus

The cowpea seed beetle, Callosobruchus maculatus, will lay its eggs on many potential hosts and inert surfaces. Oviposition on glass beads is stimulated by coating them with individual fatty acids. Nevertheless, female beetles reject mung seeds less frequently than beads treated with either an extract of mung seeds or, especially, an extract of mung seeds plus oleic acid. The addition of oleic acid to the extract resulted in a change in the sequence of oviposition behavior, notably an increase in a raised body position indicative of hosts of low acceptability. Fatty acids are present in the epicuticular waxes of legume seeds; wax extract of mung bean contains 32.4% fatty acid and 14 alkanes, whereas a wax extract of chickpea contains 5% fatty acid and 18 alkanes. Thus, chickpea may be a less acceptable host for oviposition than mung bean because of physical differences and/or because of chemical differences, including a reduced total level of fatty acid or the high proportion of oleic acid it is reported to contain. It is concluded that an appropriate mixture of fatty acids in the epicuticular waxes stimulates oviposition but that an elevated level of oleic acid in conjunction with others is deterrent.