Geographic variation of sex pheromone and mitochondrial DNA in Diatraea saccharalis (Fab., 1794) (Lepidoptera: Crambidae)

(9Z,11E)-hexadecadienal and (Z11)-hexadecenal, the main sex pheromone components of the sugarcane borer, Diatraea saccharalis, were identified and quantified from four Brazilian and one Colombian populations using GC-EAD, GC-MS and GC analyses. Three different ratios were observed, 9:1, 6:1, and 3:1. The pheromone concentration for the major component, (9Z,11E)-hexadecadienal, varied from 6.8 ng/gland to 21.9 ng/gland and from 1.7 ng/gland to 6.5 to the minor component, (Z11)-hexadecenal. The 25 D. saccharalis cytochrome oxidase II sequences that were analyzed showed low intra-specific variation and represented only 11 haplotypes, with the most frequent being the one represented by specimens from São Paulo, Paraná, and Pernambuco states. Specimens from Colombia showed the highest genetic divergence from the others haplotypes studied. Data on the genetic variability among specimens, more than their geographic proximity, were in agreement with data obtained from analyses of the pheromone extracts. Our data demonstrate a variation in pheromone composition and a covariation in haplotypes of the D. saccharalis populations studied.     

Development of the braconid wasp Cotesia flavipes in two Crambids, Diatraea saccharalis and Eoreuma loftini: Evidence of host developmental disruption

Cotesia flavipes is an important gregarious larval endoparasitoid of several crambid stem borers, including Diatraea saccharalis. The suitability of two crambid species, Eoreuma loftini and D. saccharalis, pests of sugarcane and rice in Texas, for C. flavipes development was tested. The effect of parasitization by C. flavipes on encapsulation response was assessed in vivo in both D. saccharalis and E. loftini. The results indicated that the parasitoid developed and emerged successfully in D. saccharalis larvae. Although E. loftini larvae were readily parasitized by C. flavipes parasitoids, no wasp larvae hatched from the eggs in this host because eggs were encapsulated by the host's hemocytes. The developmental fate of the E. loftini larvae with encapsulated parasitoids was variable. Most died as abnormal fifth instars or as post-wandering prepupae, while a few developed normally to the pupal stage. In vivo experiments, there was a significant reduction in the percent of beads encapsulated in parasitized larvae in both hosts. However, the percent of beads showing melanization decreased significantly in parasitized D. saccharalis larvae but did not differ significantly in parasitized or unparasitized E. loftini larvae. Our results showed that D. saccharalis is a suitable host for C. flavipes whereas E. loftini is an unsuitable host. This study indicated that lepidopteran stem borers that are taxonomically, behaviorally, and ecologically very similar can differ in their ability to encapsulate a parasitoid species.     

Structural organization of the sex pheromone gland in Helicoverpa zea in relation to pheromone production and release

Morphological location of the sex pheromone producing area in the ovipositor of the female corn earworm Helicoverpa zea, was correlated with gas chromatographic analysis of the extracted pheromone. Histological studies showed that the pheromone gland occupied an almost complete ring of specialized columnar cells between the 8th and 9th abdominal segments. Ultrastructure of the pheromone gland cells revealed distinct features such as microvilli, pockets of granular material, intercellular canals with abundant desmosomes. Apparent changes in some of these features are associated with phases of pheromone production and non-production. Examination of the tissue with low temperature scanning electron microscopy showed the presence of excreted droplets at the tips of cuticular hairs in the glandular area during the period of pheromone production.     

Discovery of a disused desaturase gene from the pheromone gland of the moth Ascotis selenaria, which secretes an epoxyalkenyl sex pheromone

Female Ascotis selenaria (Geometridae) moths use 3,4-epoxy-(Z,Z)-6,9-nonadecadiene, which is synthesized from linolenic acid, as the main component of their sex pheromone. While the use of dietary linolenic or linoleic fatty acid derivatives as sex pheromone components has been observed in moth species belonging to a few families including Geometridae, the majority of moths use derivatives of a common saturated fatty acid, palmitic acid, as their sex pheromone components. We attempted to gain insight into the differentiation of pheromone biosynthetic pathways in geometrids by analyzing the desaturase genes expressed in the pheromone gland of A. selenaria. We demonstrated that a Δ11-desaturase-like gene (Asdesat1) was specifically expressed in the pheromone gland of A. selenaria in spite of the absence of a desaturation step in the pheromone biosynthetic pathway in this species. Further analysis revealed that the presumed transmembrane domains were degenerated in Asdesat1. Phylogenetic analysis demonstrated that Asdesat1 anciently diverged from the lineage of Δ11-desaturases, which are currently widely used in the biosynthesis of sex pheromones by moths. These results suggest that an ancestral Δ11-desaturase became dysfunctional in A. selenaria after a shift in pheromone biosynthetic pathways.     

Feeding and hemolymph trehalose concentration influence sex pheromone production in virgin Heliothis virescens moths

Previously, we demonstrated that sex pheromone production in mated female Heliothis virescens moths is dependent upon hemolymph trehalose concentration (HTC), which is influenced by activities such as the feeding of adults on sucrose. In this paper we demonstrate, for the first time, that this effect also occurs in starved (i.e., sugar-stressed) virgin females. Females allowed to feed on sugar for 6 days, following eclosion, had significantly greater titers than females that had fed only on water (i.e., were starved). No differences in pheromone titer were observed between sugar- and water-fed females at shorter (1 or 3 days) periods following eclosion. The relatively short-term effects of HTC on sex pheromone titer of virgins, were demonstrated by feeding experiments, in which starved (for 4 days) virgins fed on 10% sucrose solution had significantly greater HTC and pheromone titers than ones fed only on water; an increase in HTC was apparent within an hour, while the increase in pheromone titer was apparent within 2.5 h, of sugar feeding. Starvation also showed similar effects on titers of pheromone gland fatty acids (pheromone intermediates) and HTC. Over 6 days of starvation, fatty acid titers and HTC declined gradually. After feeding on sucrose, titers of hexadecanoic, (Z)-9-hexadecanoic, (Z)-11-hexadecanoic and (Z)-9-octadecanoic, acids, as well as HTC, increased significantly 24 h later, but titers of octadecanoic and (Z,Z)-9,12-octadecanoic (linoleic) acids did not. Lepidoptera cannot biosynthesize polyunsaturated acids, but the lack of change in octadecanoic acid titer suggests this acid may not participate in pheromone biosynthesis. In addition to these short-term changes in pheromone and fatty acid production, mediated by HTC, a longer-term effect of age, regardless of HTC, on pheromone titer was observed. Overall, these results are consistent with hemolymph trehalose and glandular fatty acids acting as twin metabolite reservoirs for pheromone biosynthesis. Hemolymph trehalose, able to be refilled through feeding on exogenous sugars, has a one-way flow of metabolites for synthesis of glandular free fatty acids (FFAs) and pheromone, while glandular glycerolipids provide a reversible reservoir for metabolites, accepting surplus FFAs when glandular concentrations are high, and providing FFAs for pheromone biosynthesis when concentrations are low.     

Isolation of the sex pheromone of the moth Argyroploce leucotreta

The sex-pheromone of the false codling moth, Argyroploce leucotreta Meyr., has been isolated and its structure assigned. The extraction of the last two segments of, and of the whole, female moth gave an extract which was immediately active. The extraction of mixed populations of male and female moths only showed pheromone activity after alumina chromatography. The structure is assigned as trans-dodec-7-en-1-yl acetate on the basis of gas-liquid chromatography studies of chain length, Lemineux oxidation to pentanoic acid, hydrogenation, hydrolysis and acetylation of the neutral hydrolysis product, and the mass spectrum. The structure has been confirmed by comparison with a synthetic sample in both chemical, physical, and pheromone properties. The cis-isomer showed no pheromone activity and did not inhibit the activity of the trans form when present in a 9:1 mixture of cis-trans-isomers.     

The secretory cycle of a gland involved in pheromone production in the noctuid moth, Pseudaletia separata

The structure of the pheromone producing gland is briefly described. Examination of variations in gland volume, nucleus to cytoplasm ratio, and distribution and form of nucleic acids suggests that the secretory cycle of this gland consists of three phases. Two types of small vesicle are found and suggested as possible sites for pheromone metabolism.     

The morphology of the sex pheromone gland in Drosophila grimshawi

The morphology of a sex pheromone-producing gland found in the abdomen of Drosophila grimshawi males was studied by light and electron microscopy. This gland, consisting of two intra-anal lobes, contains cells that resemble those of other insect pheromone glands. However, in contrast to many other insect pheromone glands that release pheromone through the cuticle, cells of the intra-anal lobes secrete into a canaliculi-duct system that empties into the anal region. The liquid secretory product flows along the surface of the intra-anal lobes and is brushed onto the substrate by fingerlike projections on the lobes' surfaces.     

Characterization of oxidase(s) associated with the sex pheromone gland in Manduca sexta (L.) females

An oxidase that converts primary aliphatic alcohols into aldehydes was discovered in the cuticle of the sex pheromone gland and in the papillae anales on the tip of the abdomen of Manduca sexta females. Oxidase activity was not found in the epidermal cells of the pheromone gland where fatty acid precursors of the pheromonal aldehydes are found. This oxidase requires oxygen and water to function and appears to have a rather broad substrate specificity. The activity of the oxidase is reduced by the application of piperonyl butoxide, which also interferes with the PBAN induced production of the natural pheromone aldehydes. However, endogenous alcohols cannot be found in the pheromone gland. Thus, it is not yet clear whether or not the oxidase is involved in the terminal step of biosynthesis of the pheromone aldehydes in M. sexta females. © Wiley-Liss, Inc.

1 This article is a U.S. Government work and, a such, is in the public domain in the United States of America.

     

Correlation of housefly sex pheromone production with ovarian development

Pheromone production in the housefly was monitored during oögenesis and in ovariectomized insects by gas-liquid chromatography (GLC) and radio-GLC. The presence of vitellogenic ovaries was required for the initiation of (Z)-9-tricosene (muscalure), (Z)-9,10-epoxytricosane and (Z)-14-tricosen-10-one synthesis. Methylalkane synthesis was enhanced by developing ovaries. Insects ovariectomized within 12 hr after emergence produced no detectable amounts of (Z)-9-tricosene, C₂₃ epoxide nor C₂₃ ketone and synthesized less methylalkanes than the controls. This effect was reversed by ovary implants. When flies were ovariectomized after oviposition, synthesis of (Z)-9-tricosene, C₂₃ epoxide and C₂₃ ketone continued. Thus, initiation of the synthesis of these C₂₃ pheromone compounds required a vitellogenic ovary, but the ovary was not required to maintain synthesis.     

Behavioural responses to female sex pheromone components in Periplaneta americana

Female American cockroaches, Periplaneta americana, release two sex pheromones, periplanone-A and periplanone-B, each of which is perceived by a specialized receptor type on the male antennae. This study investigates whether male behavioural responses to both components may differ qualitatively or quantitatively. Running, upwind orientation and wing-raising were evoked by stimulation with either component. The intensity of running increased with stimulus concentration in much the same way for both substances. However, the relative effectiveness of periplanone-A was 30 times lower in behavioural assays than in the electroantennogram. Behavioural responses to periplanone-B declined faster and were more easily adapted by repeated stimulation than those to periplanone-A. It is hypothesized that periplanone-B is responsible for long distance attraction while periplanone-A influences male behaviour near the female.     

Larval esterases of the southwestern corn borer, Diatraea grandiosella: Temporal changes and specificity

Disc electrophoresis was used to examine and characterize the esterases present in the fat body, haemolymph, and midgut of last stage larvae of the southwestern corn borer, Diatraea grandiosella. Significant temporal changes were observed in the pattern of the 4 major esterases of the fat body and 3 major esterases of the haemolymph. These changing profiles presumably relate, in part, to a requirement for the degradation of juvenile hormone (JH) in preparation for metamorphosis.The binding capacity of esterases present in the larval midgut towards JH I and three JH mimics (alkyl-3,7,11-trimethyl-2,4-dodecadienoates) was also examined. The midgut of last stage nondiapausing larvae was shown to contain a carboxylesterase which bound all three JH mimics. Another esterase which bound JH I, but not the mimics, was also present. An esterase with a similar electrophoretic mobility was detected in the haemolymph and integument. Since the JH I binding esterase did not bind the JH mimics, the mimics do not appear to synergize JH by inhibiting its ester hydrolysis.     

Female sex pheromone components of the cotton bollworm, Heliothis armigera

Detailed examination of abdominal tip extracts from adult female Heliothis armigera revealed the presence of two components which elicit electroantennographic responses from the male moth. These olfactory stimulants have been fully identified as (Z)-11-hexadecenal (I) and (Z)-11-hexadecen-1-ol(II), and detected in airborne volatiles from a ‘calling’ female moth. A third olfactory stimulant was detected only in female tip extracts from some moths of Malawi origin, and this was tentatively identified as (Z)-9-hexadecenal (III). No other olfactory stimulants could be found, although hexadecenal (IV) and 1-hexadecan-1-ol (V) were detected by gas chromatography. In field tests in Malawi, (Z)-11-hexadecenal (I) attracted a few male H. armigera moths to traps but was very much less attractive than the virgin female moth. The attractiveness of (I) was not consistently affected by addition of alcohol (II), aldehydes (III) and (IV), or (E)-11-hexadecenal. Significant numbers of male Earias biplaga moths were found to be attracted to (Z)-11-hexadecenal (I).     

The female sex pheromone of the staphylinid beetle, Aleochara curtula

The grasping reaction with the parameres towards the female was used as the criterion of male sexual behaviour in Aleochara curtula. Visual, gustatory, and mechanical stimuli were excluded as triggers in releasing the reaction. Extracting the females with acetone, or covering the females with wax resin eliminated the male response. The grasping reaction was shown towards models contaminated with hexane extracts of females, whereas extracts of males were ineffective, as was the pure solvent. 0.5 female equivalents appears to give the optimal response. This is the first record of female sex pheromones acting as an aphrodisiac in the Staphylinidae.The pheromone is spread over the entire surface of the body, and the trapping of the pheromone by the epicuticular waxes is discussed. The evaporation of the pheromone from freshly-killed females is slow by comparison with that from extracts. A reduction of the free surface area of pheromone-bearing models causes a reduction in the response by the males.     

An integumentary pheromone-secreting gland in Atta sp: Territorial marking with a colonyspecific pheromone in Atta cephalotes

Workers of Atta cephalotes mark the area around their nest with a pheromone that has at least two components, one of which is colony-specific. Another, which was isolated and tested for its activity, is genus- or species-specific in its action; it appears to be similar in A. sexdens and A. cephalotes, but differs in Acromyrmex octospinosus. The pheromone is produced in a newly described gland, located near the sting. A synthetic trail pheromone component in very low concentrations stimulates some behavioural effects similar to those of the territorial pheromone.     

Temporal pattern of sex pheromone release by female Trichoplusia ni

The release rate of the pheromone component Z-7-dodecenyl acetate was determined for individual female Trichoplusia ni by using small glass tubes filled with Porapak Q to extract pheromone from air in the immediate vicinity of the everted gland. The change in release rate as a function of time was determined by taking sequential 5 min. samples from individual 4-day-old females through each pheromone release period over an entire night. The release rate was found to decline exponentially from a mean of 22 ng per min. initially to 12 ng per min. at the end of 20 min., the average length of a pheromone release period. Pheromone collections were also made from females of different ages, using a single tube of Porapak per female to collect pheromone for an entire night. The nightly mean release rate increased significantly with age, although the time spent releasing pheromone per night decreased significantly with age (to 6 days old).     

Ultrastructural changes of the posterior silk gland cells in the early larval instars of the silkworm, Bombyx mori

The posterior silk gland cells in the first three larval instars show characteristic changes during growth that are essentially similar to those undergone in the fourth larval instar. In the feeding stage, when the cells grow rapidly, vesico-tubular rough endoplasmic reticulum and a number of Golgi vacuoles occur in the cytoplasm and the glandular lumen is filled with fibrous materials, probably fibroin. In the moulting cycle when the cells stop growing, a series of degenerative changes occur such as the appearance of autophagosomes, autolysosomes, and large vacuoles. Fibrous materials disappear from the glandular lumen. These cyclic changes are discussed in relation to hormonal changes. Intercellular junctions and the tracheal system of the silk gland are described.     

Insect sex pheromone production in yeasts and plants

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Comparative binding of Cry1Ab and Cry1F Bacillus thuringiensis toxins to brush border membrane proteins from Ostrinia nubilalis, Ostrinia furnacalis and Diatraea saccharalis (Lepidoptera: Crambidae) midgut tissue

The European (Ostrinia nubilalis Hübner) and Asian corn borers (Ostrinia furnacalis Guenée) are closely related and display similar sensitivity to Cry1 toxins. In this study, we compared the binding patterns of Cry1Ab and Cry1F toxins between both Ostrinia spp., as well as the expression of putative cadherin- and aminopeptidase-N (APN)-like protein receptors. Additionally, cDNA sequences of these putative toxin receptors from both Ostrinia species were compared. Ligand blots for both species indicated a similar binding pattern for Cry1Ab with the strongest immunoreactive band at 260 kDa in both species. In addition, similar expression of the putative cadherin- and APN-like protein receptors were observed at 260 and 135 kDa, respectively. A high degree of similarity (98% amino acid sequence identity) of cDNA sequences for both putative receptor sequences was observed. The Cry1F ligand blot revealed that O. furnacalis and O. nubilalis BBMV exhibited slightly different binding patterns, with strong binding to putative proteins at 150 and 140 kDa, respectively. Both proteins appeared to also bind Cry1Ab, although the signal intensity was much reduced with Cry1Ab. O. furnacalis showed an additional but weaker band at 210 kDa relative to the 150 kDa band. Diatraea saccharalis (Fabricius), which was used as an outgroup species, exhibited different binding patterns than either Ostrinia species, with both Cry1Ab and Cry1F toxins binding to a 210 kDa protein. These results support the previous experiments indicating that O. nubilalis and O. furnacalis share similar patterns of susceptibility to Cry toxins.