Oviposition Stimulants Research Articles

Did certain chemoreceptors for host plant stimuli in the cabbage root fly evolve as pheromone receptors?

The cabbage root fly possesses highly specialised and extremely sensitive receptors for “CIF”, a group of compounds present on cabbage leaves in very small amounts, and the strongest oviposition stimulants known. Here we present evidence that the same receptors are sensitive to a methanol extract of cabbage root fly eggs, which may contain the fly's host marking pheromone. Based on these results, the possible role of CIF in cabbage root fly behaviour and in cabbage plants is discussed.

Benzoic acid glucosinolate esters and other glucosinolates from Arabidopsis thaliana

The spectacular recent progress in Arabidopsis thaliana molecular genetics furnishes outstanding tools for studying the formation and function of all metabolites in this cruciferous species. One of the major groups of secondary metabolites in A. thaliana is the glucosinolates. These hydrophilic, sulfur-rich glycosides appear to serve as defenses against some generalist herbivores and pathogens, and as feeding and oviposition stimulants to specialist herbivores. To help study their biosynthesis and role in plant-insect interactions, we wanted to determine the complete glucosinolate content of A. thaliana. In previous studies, 24 glucosinolates had been identified from ecotype Columbia. We reinvestigated Columbia as well as additional ecotypes and mutant lines, and identified 12 further glucosinolates, including five novel compounds. Structures were elucidated by MS and NMR spectroscopy of their desulfated derivatives, and by enzymatic cleavage of the attached ester moieties. Four of the novel glucosinolates are benzoate esters isolated from the seeds. In all but one of these compounds, esterification is on the glucose moiety rather than the side chain, a very unusual feature for glucosinolates. Among additional glucosinolates identified were the first non-chain elongated, methionine-derived glucosinolate from A. thaliana and the first compounds that appear to be derived from leucine.

Olfactory cues for oviposition behavior in Toxorhynchites moctezuma and Toxorhynchites amboinensis (Diptera: Culicidae).

In laboratory tests gravid female Toxorhynchites moctezuma (Dyar & Knab) and Toxorhynchites amboinensis (Doleschall) were offered the choice of black oviposition jars containing a diethyl ether extract of water collected from a natural oviposition site for these species (i.e., used tires), a dilution series of 4-methylcyclohexanol, 3-methylindole, 2-methylphenol, 3-methylphenol and 4-methylphenol, or solvent. Tire water extract and all test compounds acted as oviposition attractants and stimulants for both species, with the threshold amounts required to elicit these behaviors varying between the species and among the compounds tested.

Oviposition responses by hessian fly, Mayetiola destructor, to wheats varying in surfaces waxes.

Leaf waxes are known to contain oviposition stimulants for Hessian fly. In dual-choice tests comparing seedlings of each of three nonglaucous lines of wheat with its corresponding glaucous line, Hessian flies laid similar numbers of eggs on each genotype. However, when plants from these genotypes were tested at the flag-leaf stage. Hessian flies deposited 25-100% more eggs on the nonglaucous genotype compared to normal wax genotype in each pair. Leaf waxes extracted from the genotypes in one of these pairs (Avalon and nonglaucous Avalon) at the seedling and the flag-leaf stage and tested on paper leaf models elicited oviposition responses similar to those observed on the intact plants. Scanning electron microscopy showed differences in crystal density on nonglaucous and glaucous genotypes only at the flag-leaf stage. Gas chromatography-mass spectrometry of the leaf wax extracts showed that fly oviposition responses were associated with differences in the chemical composition of the waxes. This is the first report of Hessian fly oviposition responses varying among genotypes of wheat with different surface wax composition.

Rutabaga roots, a rich source of oviposition stimulants for the cabbage root fly

Two recently identified compounds ("CIFs"), present on the leaf surface of Brassica oleracea (cabbage), are the strongest oviposition stimulants known for the cabbage root fly, Delia radicum. Cabbage leaves contain these compounds in extremely low concentrations, and the amount of CIFs obtained from purifying leaf extracts was so small that it limited further research. We were able to purify far more of these two compounds from the roots of Brassica napus var. napobrassica (rutabaga). Apart from being a richer source of CIFs, rutabaga roots are considerably easier to collect and process than leaves. In addition, we isolated and identified a new CIF compound from the roots that is also very active in stimulating oviposition in the cabbage root fly.

Host acceptance behaviour of the small ermine moth Yponomeuta cagnagellus : larvae and adults use different stimuli

The sugar alcohol dulcitol is a strong feeding stimulant for larvae of the small ermine moth Yponomeuta cagnagellus. In this paper we tested the hypothesis that dulcitol also acts as an oviposition stimulant for this species. We found that the sugar-alcohol dulcitol was present on the surface of the host Euonymus europaeus. We also showed that (as yet unidentified compounds) can be systemically transferred (i.e. by uptake and transport via the vascular system) from E. europaeus, to the non-host Crataegus monogyna and stimulate oviposition. However, no evidence was found that this stimulatory activity was due to dulcitol. Systemic enrichment of C. monogyna with dulcitol did not induce oviposition on this plant. Neither was the application of pure dulcitol on artificial twigs effective. In addition it was shown that when dulcitol was removed from host plant extracts, oviposition stimulatory activity was retained in the fraction without dulcitol. Synergism between dulcitol and other stimulants could not be demonstrated, however, high concentrations of dulcitol in combination with the main stimulant(s) showed a trend towards enhanced oviposition. It is concluded that the sugar alcohol dulcitol can only play a limited role in adult host acceptance behaviour. The hypothesis that a host shift in the genus Yponomeuta from Celastraceae to Rosaceae might have been facilitated by the presence of small amounts of dulcitol in Rosaceae therefore needs to be restricted to larval feeding behaviour.

Identification of 1-octacosanal and 6-methoxy-2-benzoxazolinone from wheat as ovipositional stimulants for Hessian fly, Mayetiola destructor.

Bioassay-guided fractionations of a dichloromethane extract of wheat epicuticular wax allowed the identification of 1-octacosanal and 6-methoxy-2-benzoxazolinone (MBOA) as the major components that stimulate oviposition by the Hessian fly, Mayetiola destructor. These compounds were identified by their mass spectral fragmentation patterns and by comparison of their gas chromatographic retention times with synthetic samples. Synthetic samples of 1-octacosanal or MBOA stimulated significant oviposition when compared with solvent controls. In combination, these compounds elicited a synergistic effect on the number of eggs laid by females compared to when they were presented alone. In a choice bioassay, a mixture of synthetic 1-octacosanal and MBOA in the approximate concentrations determined to be present in one plant equivalent of crude extract stimulated the same amount of oviposition as one plant equivalent of extract. This showed that together these two compounds appear to be responsible for the major proportion of the ovipositional stimulatory activity of the wheat epicuticular wax extract. Comparison of the activity of five straight-chain primary aldehydes with chain lengths from C22 to C30 revealed a relationship between chain length and the number of eggs laid by female Hessian flies, with 1-hexacosanal and 1-heptacosanal the most active of the aldehydes tested.

Ovipositional Response of Tobacco Hornworm Moths (Lepidoptera: Sphingidae) to Tobacco Plants Grown Under Elevated Levels of Ozone

Tropospheric ozone (O 3 ) is an important air pollutant that can injure plants, and it may have direct or indirect effects on associated insects. Thus, tobacco plants, Nicotiana tabacum L., were exposed to 4 concentrations of O 3 in open-top chambers at the U. S. Department of Agriculture, Agricultural Research Service, Air Quality Research Unit at Raleigh, NC. The O 3 treatments were charcoal-filtered air (CF), nonfiltered air (NF), and NF with O 3 added for 12 h/d (0900-2100 hours EST) to obtain proportions of O 3 1.4 times ambient (NF1) and 1.7 times ambient (NF2). After plants had been exposed to ozone for at least 5 d, eggs from tobacco hornworm moths, Manduca sexta (L.), were counted and removed daily from the plants. Hornworm moths oviposited significantly more eggs on plants grown in the highest O 3 treatment (NF2), However, when plants were removed from the chambers, or when the O 3 was turned off in the chambers, there were no differences in ovipositional preferences of M. sexta females. Tobacco plants in treatments with enhanced O 3 had lower levels of total cuticular cembranoid diterpenes, which are important ovipositional stimulants for M. sexta. Therefore, observed changes in leaf-surface chemistry (i.e., lowered cembranoid diterpenes) do not explain the higher oviposition rates by M. sexta moths on plants grown in chambers with enhanced O 3 .

Oviposition stimulants for the cabbage root fly: isolation from cabbage leaves

Abstract Two compounds present on the surface of Brassica oleracea cv. botrytis leaves have been isolated and identified which stimulate very effectively oviposition in the cabbage root fly, Delia radicum and which are perceived by a specific receptor neuron in the tarsal sensillum C5 of the female fly. Activity of extracts and chromatographic fractions were bioassayed, using oviposition experiments and mainly electrophysiological recordings from the C5 tarsal contact chemoreceptor sensillum of female flies. Spectroscopic data indicate that the main compound is 1,2-dihydro-3-thia-4,10,10b-triaza-cyclopenta[.a.]fluorene-1-carboxylic acid, a novel compound related to Brassica phytoalexins like brassicanal C. It is accompanied by its glycine conjugate.

Composition and Variability of Epicuticular Lipids of Azaleas and their Relationship to Azalea Lace Bug Resistance

Epicuticular lipids were extracted from the foliage of six deciduous and one evergreen azalea genotypes (Rhododendron sp.) and identified by gas chromatography-mass spectrometry. The relationship of leaf-surface lipid composition with measures of resistance to azalea lace bug, Stephanitis pyrioides Scott, was evaluated. Each genotype had a distinct epicuticular lipid composition. The major surface lipid components from all test taxa were n-alkanes and triterpenoids. In the most resistant genotypes [R. canescens Michaux and R. periclymenoides (Michaux) Shinners] ursolic acid, n-hentriacontane, and n-nonacosane were the most abundant epicuticular lipids. The lipids present in largest proportion among all susceptible deciduous genotypes tested were α-amyrin, β-amyrin, and n-nonacosane. The proportions of the lipid components from the same plant of each genotype varied between spring and fall samples. Among classes of lipids, n-alkanes, n-1-alkanols, and triterpenoids had significant correlations with azalea lace bug behavior on host plants. Among individual components, heptadecanoic acid, n-hentriacontane, oleanolic acid, ursolic acid and one unknown compound (with major mass spectra 73/179/192/284/311) were significantly negatively correlated with host plant susceptibility to azalea lace bug, as measured by oviposition, leaf area damaged, egg and nymphal development, and nymphal survivorship. Triacontanol, α-amyrin, β-amyrin, and three unknowns were significantly positively correlated with host plant susceptibility. Acceptance or rejection by azalea lace bug to a particular plant may be mediated by a balance of positively and negatively interpreted sensory signals evoked by plant chemicals. This study indicated that the high levels of resistance observed in R. canescens and R. periclymenoides may be due to the lesser amount or the absence of attractants and stimulants for feeding or oviposition.

Resistance of Solanum berthaultiiFoliage to Potato Tuberworm (Lepidoptera: Gelechiidae)

Ovipositional behavior, larval establishment, and development biology of the potato tuberworm, Phthorimaea operculella Zeller, were compared on foliage of 2 accessions of wild, insect-resistant potato species, Solanum berthaultii Hawkes, and that of a popular processing potato cultivar, Atlantic (S. tuberosum L.). Total egg deposition (on and off foliage) on S. berthaultii was (approximately equal to 1/2 that on Atlantic in both free-choice and no-choice assays. In no-choice tests, only 11% of all eggs laid were deposited on foliage of S. berthaultii compared with a 92% deposition rate on foliage of Atlantic, suggesting the presence of a chemical or physical deterrent or the absence of ovipositional stimulants on foliage of S. berthaultii. A large proportion of larvae placed on Atlantic dipersed to the abaxial leaf surface and constructed a protective silk tent before initiating feeding. Only 1 neonate placed on S. berthaultii constructed a silk tent, and no neonates dispersed to the abaxial surface. Mortality of larvae confined to the foliage of S. berthaultii (accession Ber 331) was (approximately equal to 7 times greater than that of larvae on Atlantic. Leaf area consumption by individual larvae was approximately equal to 50% less on the wild species compared with cultivated potato. Larvae reared on S. berthaultii produced pupae which weighed significantly less than their siblings reared on Atlantic. Collectively, these negative impacts on potato tuberworm suggest that foliage of S. berthaultii express characteristics, that if under genetic regulation, might be useful in creating potato tuberworm-resistant potato cultivars through traditional breeding or molecular approaches.

Purification and characterization of an oviposition-stimulating protein of the long hyaline tubules in the male migratory grasshopper, Melanoplus sanguinipes

An oviposition-stimulating protein (OSP) was isolated and purified from the long hyaline tubules of the male accessory gland complex in the migratory grasshopper, Melanoplus sanguinipes. Gel filtration of the native OSP, using Sephadex G-100, indicates its molecular weight to be about 60 000 Da with the oviposition-stimulating activity while sodium dodecyl sulphate–polyacrylamide gel electrophoresis shows that the OSP comprises two subunits, each with a molecular weight of 30 000 Da. The purified OSP appears as a single symmetric peak on fast performance liquid chromatography using Mono Q. Isoelectric focusing of the OSP indicates an apparent pI of 5.5. Injection of the OSP induces oviposition in about 70% of ovulated virgin females within 48 h. Stimulation of oviposition can be blocked by a polyclonal antibody raised against the OSP 30 000 Da subunits. Amino acid analysis of the dimer and its subunits shows a comparatively high content of aspartic acid/asparagine (14.8%) as well as leucine (12.2%) and glutamic acid/glutamine (12.0%). The N-terminal 21 amino acid sequence of the OSP shows little similarity to known peptides. Immunoreactivity with the anti-OSP antibody was observed in the viscous secretion, spermatheca, and the egg-pod froth of mated females, confirming transfer of the OSP from male to female during copulation.

Oviposition Stimulant for a Gall-Inducing Sawfly, Euura lasiolepis, on Willow is a Phenolic Glucoside

The ovipositional stimulation for the shoot galling sawfly, Euura lasiolepis, whose natural host is the North American willow, Salix lasiolepis, was studied in response to its original host, Finnish willows, poplar species, and individual phenolic glucosides and fractions. The major phenolic glucosides in the natural host were salicortin and tremulacin, which were also the major components in the Finnish S. purpurea and S. rosmarinifolia. The natural host, S. lasiolepis, was chosen over six Finnish willow species in multiple-choice tests. Multiple-choice tests with six Finnish willow species and one Finnish popular species showed that S. rosmarinifolia was significantly more attractive than any of the other species by at least 16-fold. No-choice testing with phenolic glucosides showed the tremulacin fraction to be the significant ovipositional stimulant, which elicited probes with the ovipositor at least 19 times more frequently than other glucosides. A tremulacin fraction at 1.5% elicited significantly more probes than a tremulacin fraction tested at 0.4%. In addition, tremulacin was the only glucoside to induce prolonged and repeated probing with the ovipositor, lasting a mean of 6.4 sec per bout per female. Only single probes lasting a fraction of a second were observed in response to other glucosides. This is the second study showing a direct effect of a phenolic glucoside as an ovipositional stimulant for a sawfly, and it also suggests the existence of a mechanism that enables females to respond positively in a graded way to shoot length heterogeneity in host plants and populations.

Host-Plant Chemistry Influences Oviposition Choice of the Spicebush Swallowtail Butterfly

The spicebush swallowtail, Papilio troilus (L.), lays its eggs on plants in the family Lauraceae. Sassafras [Sassafras albidum (Nutt.) Nees], spicebush [Lindera benzoin (L.) Spreng.], redbay (Persea borbonia (L.)] and camphortree [Cinnamomum camphora (Nees) Eberm.] are four of its known host plants. In one-choice tests, free-flying spicebush swallowtail females laid eggs on chemical extracts of the leaves of each of these four hosts. In two-choice experiments, females always preferred to oviposit on an extract of sassafras compared to extracts of the other three hosts. It was shown for spicebush extract that this response was not due to oviposition experience. Previously we had identified one of the host plant chemicals acting as an oviposition stimulant in sassafras extract as 3-caffeoyl-muco-quinic acid (3-CmQA). Extracts of the other three hosts did not contain this compound. The addition of 3-CmQA alone to spicebush extract did not increase oviposition activity. It did, however, increase discrimination between hosts and nonhosts. When a fraction of sassafras extract containing 3-CmQA and other synergistic stimulants was added to spicebush extract, preference for sassafras extract was no longer recorded. These results show existing differences in oviposition chemistry among host plants of the spicebush swallowtail and how these differences can influence oviposition choice in bioassay experiments.

An Oviposition Stimulant for Spicebush Swallowtail Butterfly, Papilio troilus , from Leaves of Sassafras albidum

Female butterflies of the spicebush swallowtail, Papilio troilus, are specialists, ovipositing on plants in the family Lauraceae. Column chromatography and HPLC were used to isolate an oviposition stimulant from the leaves of one of its hosts, Sassafras albidum. The stimulant was identified as 3-trans-caffeoyl-muco-quinic acid on the basis of FAB-MS and 1H NMR spectra as compared to a compound previously isolated from another plant. It was not active alone, but it increased the oviposition activity of butterflies when combined with other stimulant(s) at a concentration of 7 ng/mm2 leaf surface area. Other caffeoylquinic acid isomers tested did not have this effect. This is the first report of a swallowtail contact oviposition stimulant from a plant in the family Lauraceae.

Patterns of Oviposition Stimulants for Carrot Fly in Leaves of Various Host Plants

Undamaged leaves of 12 host-plant species differing widely in acceptability to ovipositing carrot flies were extracted with a microwave-assisted method with hexane as solvent. The highly stimulatory diethyl ether fraction obtained by separation on a silica gel column was semiquantitatively analyzed by GC-MS for previously identified oviposition stimulants of the carrot fly (phenylpropenes, fluranocoumarins, polyacetylenes). Various plant species exhibited widely differing profiles of these compounds. In choice assays, moderate numbers of eggs were deposited underneath surrogate leaves sprayed with fractions that contained high amounts of just one type of compound and low amounts of the other two types. Only fractions with medium to high levels of at least two compound classes elicited strong ovipositional responses (e.g., phenylpropenes and polyacetylenes in Daucus carota, furanocoumarins and polyacetylenes in Heracleum sphondylium and Conium maculatum). None of the examined plants contained high quantities of all three compound classes. The contents of the stimulants seemed to account in a synergistic manner for the variation in activity of the diethyl ether fraction. However, they could not explain adequately the observed preference hierarchy of the carrot fly for the host-plant species.

The effect of single, double, and triple matings on the lifetime fecundity of Callosobruchus analis and Callosobruchus maculatus (Coleoptera: Bruchidae)

This study examined the effect of single, double, and triple matings on female lifetime fecundity in two closely related species of beetle, Callosobruchus analis and C. maculatus. Multiple mating resulted in elevated lifetime fecundities in both species, although the pattern of elevation differed between the two species. When oviposition resource was plentiful the elevation was apparent after a second but not a third copulation for C. maculatus and after a second and after a third for C. analis. By altering the availability of oviposition sites to C. maculatus females, we were able to alter the pattern of fecundity elevation between matings. When oviposition sites were limited, fecundity increased after a third mating but not a second mating. We suggest that the pattern of fecundity elevation associated with multiple mating in these species is primarily the result of oviposition stimulants delivered by males, rather than a nutritional “donation” brought about by the metabolism of ejaculates. We also suggest that the intra- and interspecific differences in the relationship between mating frequency and fecundity are the result of differences in egg maturation rates.

Flavanonol Glucoside and Proanthocyanidins

An oviposition stimulant for the cerambycid beetle, Monochamus alternatus, from an aqueous acetone extract of inner bark of Pinus densiflora, was identified as a flavanonol glucoside, (-)-2,3-trans-dihydroquercetin-3'-O-β-D-glucopyranoside. Two dimeric proanthocyanidins, along with polymeric proanthocyanidins, were also isolated from the extract and identified as procyanidins B-1 [epicatechin-(4 β→ 8')-catechin] and B-3 [catechin-(4α→ 8')-catechin]. In the presence of a fraction of the extract, a mixture of the polymers and either B-1 or B-3 stimulated oviposition of the female beetle. However, the glucoside, B-I, B-3 and the polymers were inactive alone.

Laboratory and field evaluations of oviposition responses of Aedes albopictus and Aedes triseriatus (Diptera: Culicidae) to oak leaf infusions.

Organic infusions created by fermenting white oak (Quercus alba L.) leaves in water were evaluated as sources of attractant odorants and contact oviposition stimulants for gravid Aedes albopictus (Skuse) and Aedes triseriatus (Say). Infusions were bioassayed in the laboratory by giving single females a choice of ovipositing in 1 container with infusion and 7 containers with water. Ae. albopictus laid significantly more eggs in containers with infusion, regardless of concentration (dilutions ranging from 10 to 100%) or age (fermentation periods of 7, 28, 60 d), than in containers holding water. The largest proportion of eggs (76.8%) was deposited in response to a 60% concentration of 7-d-old infusion. In contrast, Ae. triseriatus exhibited variable oviposition responses but generally deposited the largest number of eggs in only a few concentrations of older age infusions. In binary "sticky screen" bioassays, there was no difference between the numbers of females attracted to infusion or water, indicating that oviposition responses to infusion were mediated by contact chemostimulants rather than by attraction to odorants. Oviposition responses to infusions by field populations of Ae. albopictus and Ae. triseriatus in Raleigh, NC, were evaluated with pairs of oviposition traps, one containing infusion and the other containing water. Generally, Ae. albopictus laid significantly more eggs in ovitraps containing infusion regardless of its age (7, 28, and 60 d old) or the mass of leaves fermented (126 g = 1x or 504 g = 4x) than in water. In contrast, Ae. triseriatus deposited an equivalent number of eggs in traps containing water or 1x, 80% infusion regardless of its age; however, the oviposition response to ovitraps containing 4x, 7-d-old, 50% infusion was significant. Placement of an automobile tire behind an ovitrap did not increase the number of Ae. albopictus eggs laid in ovitraps containing 4x, 7-d-old, 50% infusion or water relative to ovitraps without a tire. Our research indicates that baiting ovitraps with oak leaf infusion would increase the sensitivity of surveillance efforts for Ae. albopictus and Ae. triseriatus.

Oviposition stimulant for the zebra swallowtail butterfly, Eurytides marcellus , from the foliage of pawpaw, Asimina triloba

We have isolated a caffeoylcyclohexane-1-carboxylic acid derivative, 3-caffeoyl-muco-quinic acid (3-CmQA), as a contact oviposition stimulant for the zebra swallowtail butterfly, Eruytides marcellus (Papilionidae), from the foliage of its primary host plant, Asimina triloba (Annonaceae). This compound alone was as active in stimulating oviposition by females as were the parent ethanolic plant extract and the host plant itself. Other tested isomers of 3-CmQA, including 5-caffeoylquinic acid (5-CQA or trans-chlorogenic acid), were inactive. We found, however, that experienced female butterflies responded strongly to host volatiles, which enhanced landing rates and hence oviposition.¶ This is the first report of an oviposition stimulant for a swallowtail butterfly of the tribe Graphiini. We found 3-CmQA to be the major caffeoylcyclohexane-1-carboxylic acid isomer in plants of the genus Asimina. These plants lack appreciable amounts of 5-CQA, which has been shown previously to be one of the oviposition stimulants for certain Rutaceae- or Apiaceae-feeding swallowtails of the related tribe Papilionini.¶ Our findings, along with earlier results from the tribes Troidini and Papilionini, suggest that responses by swallowtails to hydroxycinnamic acid derivatives as oviposition cues date back at least to the ancestor of the subfamily Papilioninae.