The cause of size-assortative mating in the leaf beetle Trirhabda canadensis (Coleoptera : Chrysomelidae)

Summary Examples of positive assortative mating by body size are abundant but its causes remain controversial. I show that size-assortative mating occurs in the chrysomelid beetle Trirhabda canadensis and I test a series of alternative hypotheses to explain how this mating pattern comes about. Results suggest that assortative mating in this beetle is due to the greater ease with which size-matched pairs can achieve intromission, and not due to size-biased skews in the availability of mates or mate choice favoring large individuals. There was no correlation between male and female elytron length (a measure of body size) at the initiation of courtship, but pairs assorted positively by size at the onset of intromission. Moreover, in the laboratory, there was a negative correlation between male and female size for pairs engaged in courtship that terminated without mating. Assortative mating was not associated with a large-male mating advantage and there was no evidence of female choice of large males. Nor was there unequivocal evidence for male choice of large females; although mating females were slightly larger and considerably heavier than solitary females, males did not differ in the frequency with which they rejected large and small females. Assortative mating in T. canadensis appeared to be caused by the lower ability of mismatched pairs to achieve intromission after an encounter, both when males were larger and when they were smaller than the female.     

Phaedon cochleariae and Gastrophysa viridula (Coleoptera: Chrysomelidae) produce defensive iridoid monoterpenes de novo and are able to sequester glycosidically bound terpenoid precursors

Summary. Larvae of the chrysomelid beetles Phaedon cochleariae and Gastrophysa viridula use iridoid monoterpenes for defense. The compounds are synthesized de novo in the glandular tissue and the reservoir. However, larvae feeding on leaves impregnated with thioglycosides of early precursors of iridoid biosynthesis such as the thioglycosides of 8-hydroxygeraniol, 8 and 9, rapidly accumulate in their defensive secretion. Thioglycosides combine a unique structural similarity to natural substrates with an exceptional chemical and biological stability against hydrolytic enzymes and can be, therefore, used to study transport phenomena of glycosides. The successful import suggests that the larvae possess, in addition to the de novo biosynthesis, the capability to sequester appropriate glycoside precursors that are transformed to iridoid monoterpenes in the reservoir. The uptake process displays a remarkable substrate selectivity, since the thioglycoside of geraniol 10 is not imported. From the two isomeric thioglycosides of 8-hydroxygeraniol, 8 and 9, the isomer 8 is preferred by a factor of ten. The data clearly support the existence of a highly selective transport system which enables the larvae to utilize plant derived terpenoid precursors in addition to their own de novo biosynthesis of iridoids.     

Prolonged mating in the milkweed leaf beetle Labidomera clivicollis clivicollis (Coleoptera: Chrysomelidae): a test of the “sperm-loading” hypothesis

Summary Female milkweed leaf beetles (Labidomera clivicollis clivicollis) frequently mate with more than one male, and pairs form mating associations which last for up to 42 h in the field. I tested the hypothesis that males remaining with females for long periods of time benefit by numerically overwhelming the sperm of their competitors. Male L.c.clivicollis copulated intermittently with females throughout an 11 hour period in the laboratory. When virgin females were allowed a single copulation, 94.3% of the sperm they received were located in the spermatheca immediately afterward. Males were not sperm-depleted, for they had large numbers of sperm available after one copulation (mean=230,000±43,200); the maximal number of sperm a male transferred to a female in 24 h was 30,500. There was a positive linear relationship between the number of sperm transferred and time up to 24 h after mounting (r ²=0.178, P<0.003). These data suggest that males transfer increasing numbers of sperm throughout a 24-h-period. Mating duration was the most important determinant of paternity when females were placed with one male for 24 h and another male for 6 hours. Females whose first matings were longer showed first male sperm predominance (as determined by starch-gel electrophoresis), while females whose second matings were longer showed last male sperm predominance. In view of these data, it is puzzling that males do not inseminate with large numbers of sperm immediately after mounting the female. It is possible that female refractory behaviors make insemination difficult and favor prolonged mating by male milkweed leaf beetles.     

The effect of multiple mating on genetic relatedness in larval aggregations of the imported willow leaf beetle (Plagiodera versicolora,Coleoptera: Chrysomelidae)

Summary The effect of multiple mating on the average genetic relatedness within socially interacting sib groups was studied in the imported willow leaf beetle. Using an electrophoretically determined genetic marker, it was demonstrated that between 49 and 96% of wild caught females produce egg clutches expressing multiple sires. Following theoretical predictions of Wade (1982), this information was used to estimate the harmonic mean number of sires per family and then the average relatedness within families. Average relatedness was estimated to be 0.38. An alternate form of data analysis suggested by Pamilo and Crozier (1982) did not require any knowledge of paternity and resulted in a slightly higher estimate of relatedness, 0.44.     

Predicting the acute lethality of a chlorinated benzene mixture in soils: a comparison of solid-phase microextraction,body residue,and solvent extraction techniques

When predicting the adverse effect(s) of chemical pollution on an exposed organism, a measurement of body residue is often considered a more accurate estimate of dose than those estimates derived from a solvent extraction or solid-phase microextraction of the polluted exposure medium. The magnitude of this perceived difference in accuracy, and thus the accuracy of the corresponding toxicity prediction, were investigated in the present two-phase study. In Phase I, an acute lethality response was correlated with dose estimates derived from body residue, solvent extraction, and SPME. In Phase II, acute lethality was predicted from Phase I data. The accuracy of Phase II toxicity predictions was determined by comparing predicted mortality to observed mortality. Although this study revealed that body residue indeed provided the most accurate prediction of toxicity, the margin of this comparatively greater accuracy suggests that alternative methods may provide equally viable dose estimates and, subsequently, equally viable toxicity predictions.     

Volatile emissions from the flea beetle Altica litigata (Coleoptera: Chrysomelidae) associated with invasive Ludwigia hexapetala

The flea beetle Altica litigata (Chrysomelidae) is an insect herbivore to plants within the families Lythraceae and Onagraceae, including ornamentals such as crape myrtle, Lagerstroemia spp. This insect is important both as a pest species and as a naturally occurring biological control agent due to its aggregate feeding behavior, which typically results in severe defoliation of the host plant. Despite the negative economic impact to ornamentals and contrary benefits as a biological control agent, there are few reports on the semiochemical communication of this family of insects. Uruguayan primrose-willow (Ludwigia hexapetala) is an invasive aquatic weed in California and serves as a host to A. litigata. To better characterize this association, the volatile emissions of A. litigata were collected while the flea beetles were: in containers by themselves, in containers with L. hexapetala leaves, in situ on L. hexapetala leaves in a growth chamber, and in situ on L. hexapetala leaves in the field. For comparison, the volatile emissions of A. litigata associated with two subspecies of creeping water primrose (L. peploides) were also evaluated. Two himachalene-type sesquiterpenes, showing the same carbon skeleton as compounds previously reported from Aphthona flava and Epitrix fuscula, were detected as volatiles from A. litigata.     

Behavior and ecology of Old World Luperini beetles of the genusAulacophora (Coleoptera: Chrysomelidae)

Summary The attraction of Old World leaf beetles in the genusAulacophora to kairomones and parakairomones which are effective lures for New WorldDiabrotica andAcalymma were investigated. Beetles captured on sticky traps baited with single and multicomponent lures were no different from the control traps for two species ofAulacophora. Yellow colored traps and squash blossoms are attractive toAulacophora beetles which detect sub-microgram quantities of cucurbitacins on silica gel. Leaf feeding behavior and flight activity data are correlated with varietal preference of threeAulacophora species. The common response byDiabrotica andAulacophora to cucurbitacins reinforces the two groups' coevolutionary association with the Cucurbitaceae. The apparent lack of a common response toCucurbita blossom volatiles suggests recent evolutionary pathways are substantially different for these two groups of beetles.     

Alternative reproductive tactics and male-dimorphism in the horned beetle Onthophagus acuminatus (Coleoptera: Scarabaeidae)

Adult dung beetles (Onthophagus acuminatus) exhibit continuous variation in body size resulting from differential nutritional conditions experienced during larval development. Males of this species have a pair of horns that protrude from the base of the head, and the lengths of these horns are bimodally distributed in natural populations. Males growing larger than a threshold body size develop long horns, and males that do not achieve this size grow only rudimentary horns or no horns at all. Previous studies of other horned beetle species have shown that horned and hornless males often have different types of reproductive behavior. Here I describe the mating behaviors of the two male morphs of O. acuminatus during encounters with females. Females excavate tunnels beneath dung, where they feed, mate and provision eggs. Large, horned males were found to guard entrances to tunnels containing females. These males fought with all other males that attempted to enter these tunnels. In contrast, small, hornless males encountered females by sneaking into tunnels guarded by other males. In many instances, this was accomplished by digging new tunnels that intercepted the guarded tunnels below ground. Side-tunneling behavior allowed sneaking males to enter tunnels beneath the guarding male, and mate with females undetected. Both overall body size and relative horn length significantly affected the outcome of fights over tunnel ownership. These results suggest that alternative reproductive tactics may favor divergence in male horn morphology, with long horns favored in males large enough to guard tunnels, and hornlessness favored in smaller males that adopt the “sneaking” behavioral alternative. Received: 12 October 1996 / Accepted after revision: 8 August 1997     

Chemical defense in Platyphora kollari Baly and Leptinotarsa behrensi Harold (Coleoptera: Chrysomelidae). Hypotheses on the origin and evolution of leaf beetles toxins

Summary. The defensive secretions of Platyphora kollari beetles (Chrysomelidae) from Brazil contained one oleanene glycoside. This was identified as 3-O-g-D-glucopyranosyl-(l̆)-g-D-glucuronopyranosyl-hederagenin (8), by a combination of 1D and 2D NMR methods (COSY, HMQC, MBC) and SIMS. There were also several amino acid derivatives, including the known non-protein amino acid (Z)-2-amino-3,5-hexadienoic acid (1), the new dipeptide L-n-glutamyl-2-amino-(3Z)-hexenoic acid (4), ethanolamine (7), and a mixture of phosphatidylcholines. Compounds 1 and 7 have already been identified in several leaf beetle taxa. Secretions of Leptinotarsa behrensi contained as major triterpene glycoside 3-O-g-d-glucopyranosyl-(l̅)-g-d-glucuronopyranosyl-oleanolic acid-28-O-g-d-glucopyranoside (9), already isolated from Platyphora opima. Although the host plants of these beetles were devoid of these triterpene glycosides, g-amyrin was shown to be present in three plants on which different species of triterpene-producing chrysomelids had been fed (Mikania micrantha, Ipomoea batatas and Convolvulus arvensis). This suggests that these insects are able to use g-amyrin from their food plant as a precursor to their oleanolic- and hydroxyoleanolic acid glycosides. The distribution of toxins already identified in leaf beetles suggests that amino acid derivatives could be a plesiomorphic character, considering their wide distribution, whereas the secretion of triterpene saponins, probably derived from ubiquitous plant triterpenes, could be an apomorphic character shared by some Doryphorina. The secretion of cardenolides derived from ubiquitous plant phytosterols would be another apomorphic character shared by other Doryphorina and Chrysolinina.     

Offensive and defensive sperm competition roles in the dung beetle Onthophagus taurus (Coleoptera: Scarabaeidae)

Sperm competition is predicted to generate opposing selection pressures on males. On one hand, selection should favour ‘defensive’ adaptations that protect a male’s ejaculate from displacement, while, on the other hand, selection should favour ‘offensive’ adaptations that overcome paternity assurance mechanisms of rivals. Here, we use the sterile male technique to assess sperm precedence when a male dung beetle Onthophagus taurus mates in both a defensive (first male) and an offensive (second male) role. Significant variation in a male’s sperm precedence (both P ₁ and P ₂) was detected, and an individual’s defensive (P ₁) and offensive (P ₂) abilities were positively correlated. Thus, it appears that sexual selection simultaneously selects for ‘defensive’ and ‘offensive’ adaptations in O. taurus. We discuss a variety of male traits in O. taurus that potentially contribute to a male’s ability to be successful when mating in an ‘offensive’ and a ‘defensive’ role.     

Metabolism of neolignans from P. regnellii (Piperaceae) in the beetle Naupactus bipes (Coleoptera: Curculionidae)

Piper regnellii (Piperaceae) contains high levels neolignans with diverse biological activities, including insecticidal. Despite the insecticidal activity of these neolignans, Naupactus bipes (Coleoptera: Curculionidae) larvae and adults were found feeding on the leaves (adults) and roots (larvae) of P. regnellii. The present study investigated the metabolic pathway of neolignans from P. regnellii leaves in the beetle N. bipes, focusing on possible biotransformation or sequestration of these compounds by the beetle. Two of the four plant neolignans could be recovered in the feces of adults feeding on P. regnellii leaves. In addition, four degradation products were detected, including the neolignan 3-[(2R,3R)-2,3-dihydro-3-(methyl)-2-(4-hydroxyphenyl)-5-benzofuranyl]-(2E)-propenal (7), an oxidation product of conocarpan, the major plant neolignan. The adult beetle sequesters the neolignan conocarpan selectively from leaves as well as accumulates neolignan 7. Neolignan 7 was also detected in the larvae of lab-reared beetles.     

Aggregation pheromone compounds of the black larder beetle Dermestes haemorrhoidalis Kuster (Coleoptera: Dermestidae)

Gas chromatography with simultaneous flame ionisation and electroantennographic detection (GC–EAD) and gas chromatography with mass spectrometry analysis (GC–MS) of abdominal extracts of adult male Dermestes haemorrhoidalis Kuster (Coleoptera: Dermestidae) revealed the presence of electrophysiologically and behaviourally active compounds to its conspecific males and females. Isopropyl dodecanoate (3), isopropyl (Z)-9-tetradecenoate (5), isopropyl tetradecanoate (6), isopropyl (Z)-9-hexadecenoate (7) and isopropyl hexadecanoate (8) were detected in male abdominal extracts only. Analysis of collected male headspace volatiles revealed the presence of six EAD-active compounds (3), (5), (6) and isopropyl tridecanoate (4) plus two unidentified compounds (1) and (9). Synthetic compounds (3), (4), (5), (6) and (7) showed EAD activity with antennae of both sexes in contrast to synthetic (8) which showed EAD activity with female antennae only. Male and female antennae of D. haemorrhoidalis reacted with high receptor potentials to isopropyl (Z)-9-dodecenoate (2), although this compound itself was detected in neither male nor female abdominal extracts or headspace volatiles. Petri dish bioassays indicated that male abdominal extracts and compounds (2), (3), (5) and (6) aroused and attracted conspecific male and female beetles significantly (P < 0.05) compared to female extracts. These results suggested the presence of a male-produced aggregation pheromone in D. haemorrhoidalis. Field assays with any of the described compounds, however, did not result in attraction of this beetle in significant numbers.     

Faecal shield chemical defence is not important in larvae of the tortoise beetle Chelymorpha reimoseri (Chrysomelidae: Cassidinae: Stolaini)

The most frequently investigated defence mechanism among larvae of tortoise beetles is protection by faecal shields, which generally present chemicals that are directly sequestered and/or modified from larval host-plants. In the present work we investigate the tortoise beetle Chelymorpha reimoseri that feeds on the leaves of Ipomoea carnea fistulosa (Convolvulaceae), seeking for the importance of this mechanism to their larvae. We show that 4th instar larvae suffer low predation regardless of the presence of shields in field and laboratory bioassays with ants and chicks. Chloroform extract from larvae without shields provided high protection against predation as shown in bioassays in the field, as well as against chicks, suggesting that C. reimoseri does not rely on the shield for protection. The aqueous extract of the shields did not show activity in such bioassays. The compounds responsible for this protection have yet to be identified, and it remains an open question as to whether they are sequestered from the host plant or de novo biosynthesized. This is the first record of chemical defence in cassidine beetles without the need for faecal shields. These findings indicate that more attention should be paid to chemicals present in the tissues of larvae and/or adults of tortoise beetles; the protective compounds sequestered from host plants or de novo biosynthesized can provide an alternative or complementary strategy against predation in these insects.     

Proximal traits and mechanisms for biasing paternity in the red flour beetle Tribolium castaneum (Coleoptera: Tenebrionidae)

A comprehensive understanding of sexual selection requires knowledge of the traits and mechanisms responsible for increasing a male’s paternity share (proportion of progeny sired) relative to that of other males mating with the same female. In this study we manipulated by starvation the expression of traits that might influence male paternity share in Tribolium castaneum. We then conducted experiments to examine how male starvation affects male performance during sequential episodes of sexual selection from mating to progeny production, and investigated female control over specific stages by using live vs dead females. Comparison of starved vs fed males revealed that T. castaneum females have control over spermatophore transfer during mating, as live females rejected inseminations by starved (“low quality”) males. None of the measured male copulatory behaviors (leg-rubbing frequency, asymmetry, and percent of time spent rubbing) affected the probability of successful insemination, but the last two were positively associated with male paternity share. Spermatophore positioning within the female reproductive tract was not affected by male treatment (starved/fed), by female treatment (live/dead), or by male copulatory behaviors. Starvation, however, had a dramatic effect on male reproductive physiology, decreasing both accessory gland size and total number of sperms transferred (but not sperm viability in seminal vesicles). In addition, females who mated to starved males stored fewer sperms in their spermathecae, which, together with decreased ejaculate size, may explain the reduced paternity share of starved males compared to fed males. This study elucidates some cryptic mechanisms influencing male reproductive success and aids our understanding of trait evolution through sexual selection.     

Profiles of cuticular hydrocarbons mediate male mate choice and sexual isolation between hybridising <Emphasis Type="Italic">Chrysochus</Emphasis> (Coleoptera: Chrysomelidae)

Summary. Chemical signals frequently underlie sexual isolation between insect species. Our understanding of the evolutionary forces influencing these signaling systems is known for very few systems, challenging both our efforts to understand insect speciation, and our ability to predict long-term changes in the chemical communication systems of insects. Thus, we are in need of more systems in which both the chemical signals causing sexual isolation and the evolutionary forces driving sexual isolation are understood. Sexual isolation in the hybrid zone between Chrysochus cobaltinus and C. auratus has apparently increased in response to natural selection against hybridisation (i.e. reinforcement). Previous experiments suggested that this isolation was due, at least in part, to male preferences for conspecific females. Here, we confirm this role of male choice, and document that male mate choice in this system is influenced by cuticular hydrocarbon (CHC) profiles. Specifically, male C. cobaltinus responses to control cadavers and conspecific female cadavers painted with different cuticular hexane extracts, together with analyses of the composition of those extracts, revealed that male mate choice is governed by CHC profiles. Multivariate analyses of GC profiles demonstrated that those profiles are indeed both sex- and species-specific. Although GC-MS enabled identification and quantification of the specific cuticular hydrocarbons, we have not yet determined which individual compounds govern mate choice. Having established that CHCs influence sexual isolation in this system, we can now assess the evolutionary lability of these cues, which will inform both our understanding of speciation, and of the conditions under which the chemical signaling systems that influence mate choice in insects can evolve.     

Patterns of parental provisioning covary with male morphology in a horned beetle (Onthophagus taurus) (Coleoptera: Scarabaeidae)

Male dung beetles, Onthophagus taurus, are dimorphic for a secondary sexual trait, head horns. Horned males participate in the production of brood masses while hornless male do not. Here we examine the reproductive performance of females mated with males exhibiting alternative horn morphologies. We found that exposure to males may be costly for females in that it reduced the total number of brood masses produced. However, females paired with horned males produced significantly larger brood masses than females paired with hornless males or females producing broods alone. We discuss the possible selection pressures that may underly horn evolution in this genus. Received: 22 August 1997 / Accepted after revision: 19 January 1998