Sound production in Scolytidae: Rivalry and premating stridulation of male Douglas-fir beetle

The previously reported attractant chirp of male Dendroctonus pseudotsugae changed to a distinctive long interrupted chirp after several minutes in the female gallery, and the same chirp occurred during fighting with another male. It was experimentally evoked by synthetic 3-methylcyclohex-2-en-1-one and frontalin, both known pheromone components of the male. A different, short, uninterrupted male chirp emitted shortly before copulation, was not evoked by these pheromones. It is suggested that chemostimulus allows or maintains dual contexts for the interrupted chirp. Observation of premating and mating behaviour showed some initial male aggressiveness with the female that subsided as gallery cooperation was established before mating and as the uninterrupted chirp replaced the interrupted chirp.     

(R)-Desmolactone Is a Sex Pheromone or Sex Attractant for the Endangered Valley Elderberry Longhorn Beetle Desmocerus californicus dimorphus and Several Congeners (Cerambycidae: Lepturinae)

We report here that (4R,9Z)-hexadec-9-en-4-olide [(R)-desmolactone] is a sex attractant or sex pheromone for multiple species and subspecies in the cerambycid genus Desmocerus. This compound was previously identified as a female-produced sex attractant pheromone of Desmocerus californicus californicus. Headspace volatiles from female Desmocerus aureipennis aureipennis contained (R)-desmolactone, and the antennae of adult males of two species responded strongly to synthetic (R)-desmolactone in coupled gas chromatography-electroantennogram analyses. In field bioassays in California, Oregon, and British Columbia, traps baited with synthetic (R)-desmolactone captured males of several Desmocerus species and subspecies. Only male beetles were captured, indicating that this compound acts as a sex-specific attractant, rather than as a signal for aggregation. In targeted field bioassays, males of the US federally threatened subspecies Desmocerus californicus dimorphus responded to the synthetic attractant in a dose dependent manner. Our results represent the first example of a “generic” sex pheromone used by multiple species in the subfamily Lepturinae, and demonstrate that pheromone-baited traps may be a sensitive and efficient method of monitoring the threatened species Desmocerus californicus dimorphus, commonly known as the valley elderberry longhorn beetle.     

Sound production in Scolytidae: Stridulation by female Dendroctonus beetles

A new stridulatory apparatus on the last sternite and the eighth segment is described in six female Dendroctonus bark beetles, and elytral files similar to those of males are found on four of these species, which were hitherto believed to be silent. Electronically recorded sounds include an infrequent ‘spontaneous’ click from females alone and rapidly repeated clicks (D. pseudotsugae and D. rufipennis) or a multi-impulse chirp (D. brevicomis and D. ponderosae) from females in close acoustic range of other females. An intraspecific spacing function during gallery site selection is suggested.     

Female response song in the ephippigerines Steropleurus stali and Platystolus obvius (Orthoptera,Tettigoniidae)

Stridulation by females of Steropleurus stali and Platystolus obvius in response to the calling song of the males was observed and recorded. The response has only been stimulated by the appropriate male song, either directly or from a recording. The structure of the files and the form of stridulation in both sexes is described. The male song of S. stali is remarkable in that only a few teeth on the file are struck in each wing movement. It is also notable that both opening and closing wing strokes contribute more or less equally to the syllable. The female song is similar but distinct. The song of P. obvius male is a single chirp involving nearly all the teeth on the file and with the main emphasis on the closing syllable. The response song of the female is a very brief chirp. These species are only sporadic singers, but when the female responds they are stimulated into greater activity. They thus contrast with reiterative singers like Ephippiger in which there is no female response. The implications of these contrasting behavioural patterns is discussed.     

Nematode parasites and associates of Dendroctonus spp. and Trypodendron lineatum (Coleoptera: Scolytidae), with a description of Bursaphelenchus varicauda n.sp.

Nematode parasites and associates of four bark beetle species in British Columbia were surveyed. Bursaphelenchus varicauda n.sp., Ektaphelenchus macrostylus, Panagrolaimus dentatus, and Cryptaphelenchus latus were found associated with Dendroctonus pseudotsugae. Parasitorhabditis obtusa was found in the gut and Contortylenchus reversus in the hemocoel of both D. pseudotsugae and D. rufipennis. The latter also had hemocoel infections of Sphaerulariopsis dendroetoni, which were not found concomitant with C. reversus infections. Contortylenchus brevicomi occurred in the hemocoel of D. brevicomis. The first report of a tylenchid larva parasitizing Trypodendron lineatum in North America is presented. Bursaphelenchus varicauda n.sp. was obtained from the gallery frass of D. pseudotsugae. It resembles B. corneolus and B. bestiolus but differs from the former species in female tail shape, the position of the excretory pore, spicule shape, and position of the male caudal papillae, and from the latter species in spicule shape and in the length of the esophagus and postuterine sac.     

Mating Effect on Sex Pheromone Production of the Oriental tobacco budworm,Helicoverpa assulta

This study was undertaken to clarify the suppression phenomenon of sex pheromone production after mating and its relationship to the physiological mechanism in adult females of Helicoverpa assulta, and determine the mating factor from males causing depletion of sex pheromonc production. Sex pheromone production of H. assulta females was mostly terminated in 3 hours after mating. Mated females maintained with a low titer of sex pheromone until 3 days when it started to increase again, which showed a characteristic of species mating more than once. The mated female again produced pheromone upon injection of pheromone biosynthesis activating neuropeptide (PBAN) or extracts of brain-suboesophageal ganglion complexes (Br-Sg) of mated female, which were shown similar pheromonotropic activities as compared with virgin females. These results indicated that the mating did not inhibit the receptivity of pheromone gland itself and PBAN biosynthesis in suboesophageal ganglion of the mated females. And it seems to support that the depletion of sex pheromone production is responsible for blocking of PBAN release from head. To investigate the mating factor from adult males, when extracts of reproductive organs of male were injected into hemocoel of virgin females evoking depletion of sex pheromone production as shown in mated female. The results suggest that a chemical substance(s) from the male reproductive organs could be responsible for the loss of sex pheromone biosynthesis in H. assulta.     

Sound production in scolytidae: 'rivalry' behaviour of male Dendroctonus beetles

Intrasex response of two males at the gallery of an attractive female included ‘rivalry’ stridulation that was distinct in certain acoustic properties from chirps evoked by the female attractant. With the southern pine beetle, Dendroctonus frontalis, comparison of chirps evoked by natural and synthetic chemostimulus confirmed that myrtenol is part of the female attractant, that verbenone is multifunctional according to its concentration, and that endo-brevicomin has a rivalry function as well as its anti-aggregative effect. Considerable acoustic specificity was apparent in tested chirps of three bark beetles.     

Linyphid spider courtship: Releaser and attractant functions of a contact sex pheromone

Experiments with male bowl-and-doily spiders (Frontinella communis) on methanol-washed and unwashed female webs indicate that the courtship of males is chemically released. Parallel experiments on the webs of females of another linyphiid species and on immature F. communis webs indicate that the releaser pheromone is species specific and that it is produced only by mature females. The pheromone that releases male courtship also functions as an attractant, changing the male's orientation from negatively to positively geotactic. The presence or absence of a female on the test web has only a slight effect on the male's behaviour.     

Responses of Second-Instar Male Nymphs of Four Mealybug Species (Hemiptera: Pseudococcidae) to Conspecific and Heterospecific Female Sex Pheromones

The response of the late second-instar male nymphs of the mealybug species (Hemiptera, Pseudococcidae), Planococcus citri (Risso), Planococcus ficus (Signoret), Pseudococcus cryptus Hempel Nipaecoccus viridis (Newstead), to their conspecific and heterospecific female pheromone was studied. Males that exhibited the typical appearance of late-second-instar nymphs were tested. The male behavior was monitored soon after their exposure to the tested female sex pheromone in glass Petri dish arenas. Male nymph behavior toward the pheromone source was characterized based on their aggregation on the disks in the arena. Males of all four tested mealybug species were attracted to their conspecific female pheromone. By contrast, almost no interceptions of male nymphs with disks impregnated with a heterospecific female pheromone were observed. The mode of attraction of each of male nymphs of P. ficus, among most of the tested individuals (>80%), to the conspecific female sex pheromone, (S)-lavandulyl senecioate and or (S)-lavandulyl isovalerate, was the same as the mode of attraction latter on as adult. We suggest that by being attracted to the conspecific pheromone these males may direct themselves to a suitable pupation site near conspecific non-sibling mature females, thus preventing inbreeding. The repellency of heterospecific sex pheromone to males that are looking for a pupation site suggests that the latter try to avoid close contact with heterospecific females.     

Chemical ecology and insect conservation: optimising pheromone-based monitoring of the threatened saproxylic click beetle Elater ferrugineus

Elater ferrugineus is a saproxylic click beetle inhabiting old deciduous trees in Europe. It is threatened throughout its area of distribution due to habitat loss. No efficient monitoring method has been available for this species, but observed attraction of females to (R)-(+)-γ-decalactone, which is a male-produced sex pheromone of its prey, the scarab beetle Osmoderma eremita, has led to the development of an odour lure for monitoring. In addition, four esters have recently been identified from the pheromone-producing gland in female E. ferrugineus, and a blend of these esters is highly attractive to conspecific males in the field, revealing an alternative odour-based method for monitoring this species. However, no rigorous analysis has been performed to check whether all four esters show biological activity in male E. ferrugineus, and whether its own sex pheromone is a more potent lure than the prey kairomone for monitoring of E. ferrugineus. In this study, we reinvestigated the E. ferrugineus sex pheromone, using electrophysiological and behavioural analyses, and found that only one of the esters, 7-methyloctyl (Z)-4-decenoate, is active. In addition, trapping experiments revealed that 7-methyloctyl (Z)-4-decenoate is a much more efficient attractant for male E. ferrugineus than the prey pheromone is for conspecific females, or any sex of O. eremita. With a very efficient odour lure at hand, novel information about current distribution, local population sizes, and dispersal ranges in E. ferrugineus can now be obtained, which can aid in conservation efforts to protect this threatened insect and its habitat.     

Sex pheromone of the cabbage looper: Reactions with antennal proteins in vitro

In the presence of (Z)-7-dodecen-1-ol acetate, the sex attractant of the cabbage looper, Trichoplusia ni, soluble protein from male antennae showed a time-dependent difference spectral absorbance at 280 nm. The change was associated with the enzymatic conversion of the pheromone to (Z)-7-dodecen-1-ol, a potent inhibitor of behavioural responses to the pheromone. In contrast, the response obtained with the inhibitor was indicative of non-enzymatic binding to specific protein(s) in the fraction. GLC analyses of the relative rates of enzymatic hydrolysis of the pheromone by the antennae, haemolymph, and legs revealed 33·9, 10·1, and 6·5 per cent conversion per hour, respectively. These results may provide an insight into the fate of a pheromone in the olfactory process of this insect; however, the significance of the reaction with the inhibitor is not yet known.     

Female sex pheromone of a lichen moth Eilema japonica (Arctiidae, Lithosiinae): Components and control of production

Seven candidates for components of the female sex pheromone of Eilema japonica (Arctiidae, Lithosiinae) were detected in an extract of pheromone glands with a gas chromatograph-electroantennographic detector. The compounds were identified as (Z,Z)-6,9-icosadiene (D20), (Z,Z)-6,9-henicosadiene (D21), (Z,Z,Z)-3,6,9-henicosatriene (T21), (Z,Z)-6,9-docosadiene (D22), (Z,Z,Z)-3,6,9-docosatriene (T22), (Z,Z)-6,9-tricosadiene (D23), and (Z,Z,Z)-3,6,9-tricosatriene (T23). Assays using synthetic lures in a wind tunnel showed that D21 (proportion, 0.39), T21 (0.08), D22 (0.27), and T22 (0.26) are important for evoking full behavioral responses from the males. Titers of the pheromone components did not show clear temporal fluctuations. Moreover, decapitation of the female moth had no effect on the titers of pheromone components in the pheromone gland, suggesting that cephalic endocrine factors such as pheromone biosynthesis activating neuropeptide (PBAN) are not involved in the control of pheromone biosynthesis in this species.     

Sex pheromone titre in the glands of Spodoptera litura females: circadian rhythm and the effects of age and mating

The present study investigates the effects of age and mating status on the circadian variations of gland sex pheromone titre in female Spodoptera litura Fabricius. Similar to other nocturnal moths, S. litura females exhibit circadian variations of gland sex pheromone contents, with higher levels during scotophase and lower levels during photophase. The sex pheromone titre in the glands peaks during the first scotophase after eclosion and sharply declines afterwards. Higher pheromone contents during scotophase may facilitate female reproductive activities, and the negative relationship between pheromone titre and female calling is likely the result of pheromone release during female calling. Interestingly, the present study demonstrates that mated S. litura females have significantly higher sex pheromone titre in their pheromone glands (PGs) than virgin females. This finding contrasts with all previous studies of other insect species, in which mating generally reduces the sex pheromone titre in female PGs. In S. litura, mating and male accessory gland fluids can suppress female calling behaviours and re‐matings. These results suggest that the suppression of female calling behaviours by mating and male accessory gland fluids may significantly reduce the release of sex pheromones and thus result in higher sex pheromone titre in the PGs of mated females.     

Stridulation and Courtship Behaviour of Four Endemic Pill-Millipedes, <Emphasis Type="Italic">Arthrosphaera</Emphasis> spp. (Sphaerotheriida: Arthrosphaeridae) of the Western Ghats of India

Mating behaviour of four species of pill-millipedes under genus Arthrosphaera Pocock (Arthrosphaera dalyi Pocock and Arthrosphaera disticta Pocock, Arthrosphaera fumosa Pocock and Arthrosphaera magna Attems) endemic to the Western Ghats of Southern India was analyzed in mesocosms. Stridulation is a classical communication signal in males as well as females for mate selection. Conglobation (or volvation) is a mechanism of defence to protect from disturbance or avoid predation. If male touches female or vice versa they conglobate. To avoid disparity among individuals of the same species, volvating pill-millipedes evolved stridulation behaviour for communication. The male broadcasts appropriate signal to female through stridulation to advertise its interest in mating. The females test the male’s fitness by conglobation and suitable male uncoils the partner through stridulation signals. Male with its pygidium successfully uncoils the female and attains suitable orientation for courtship. Male pairs with female ventro–ventro contact in opposite direction to deposit sperm into the vulva of female. The duration of mating varies from species to species and usually a lapse from 3 to 30 min. Vibration generated by stridulation is species-specific and its perception mechanism in pill-millipedes is yet to be clearly understood. Present study emphasized the structure of stridulatory organs, mechanism of stridulation and pattern of mating behaviour in four species of pill-millipedes.     

Gall midge olfaction: Pheromone sensitive olfactory neurons in Contarinia nasturtii and Mayetiola destructor

This study describes the morphology and function of the antennal sensilla in two gall midge species, Contarinia nasturtii and Mayetiola destructor, where multi-component sex pheromones have been identified. Both species possess sensilla trichodea, s. coeloconica, s. chaetica and s. circumfila. Sensilla circumfila, which consist of several sensilla that bifurcate and fuse into one structure, are unique for the gall midges. In C. nasturtii s. circumfila are sexually dimorphic. In males, they form elongated loops suspended on cuticular spines, whereas in females they run like worm-like structures directly on the antennal surface. Single sensillum recordings demonstrated that olfactory sensory neurons housed in male s. circumfila in C. nasturtii responded to the female sex pheromone. In M. destructor, s. circumfila were attached to the antennal surface in both sexes, and displayed no response to sex pheromone components.A sexual dimorphism was also found in the number of s. trichodea per antennal segment in both C. nasturtii (male 1 vs. female 7) and M. destructor (male 13 vs. female 10). OSNs located in male M. destructor s. trichodea responded to the sex pheromone. This is the first gall midge single sensillum study, and the first demonstration of the functional significance of s. circumfila.     

Mechanisms of pheromone inactivation in the gut of Periplaneta americana

The sex attractant of the cockroach, Periplaneta americana, has been shown in earlier work to be largely inactivated by dissected midgut from males and from mated females. It is only slightly inactivated by the midgut from virgin females. In this paper, the sex pheromone inactivating system is further studied and shown to be active in late instar larvae of both sexes. In the male this pheromone inactivation is inhibited by piperonyl butoxide, a microsomal oxidase inhibitor. This compound appears to act on the midgut tissue directly. In the mated female, piperonyl butoxide has little effect. When the pheromone inactivating capacity is partitioned into soluble and tissue components, it appears that the soluble component is most active in the male, whereas the tissue component is most active in the female. Evidence from heat inactivation, trichloracetic acid precipitation, and the use of soy bean trypsin inhibitor, as well as the time course of the reaction, suggest that the factor or factors inactivating pheromone are proteins, probably enzymes. Evidence that at least part of the pheromone inactivating capacity is due to microsomal oxidases is considered. It is also observed that both pheromone and piperonyl butoxide absorb to membranes.     

Moths behaving like butterflies. Evolutionary loss of long range attractant pheromones in castniid moths: a Paysandisia archon model

Background

In the course of evolution butterflies and moths developed two different reproductive behaviors. Whereas butterflies rely on visual stimuli for mate location, moths use the ‘female calling plus male seduction’ system, in which females release long-range sex pheromones to attract conspecific males. There are few exceptions from this pattern but in all cases known female moths possess sex pheromone glands which apparently have been lost in female butterflies. In the day-flying moth family Castniidae (“butterfly-moths”), which includes some important crop pests, no pheromones have been found so far.

Methodology/Principal Findings

Using a multidisciplinary approach we described the steps involved in the courtship of P. archon, showing that visual cues are the only ones used for mate location; showed that the morphology and fine structure of the antennae of this moth are strikingly similar to those of butterflies, with male sensilla apparently not suited to detect female-released long range pheromones; showed that its females lack pheromone-producing glands, and identified three compounds as putative male sex pheromone (MSP) components of P. archon, released from the proximal halves of male forewings and hindwings.

Conclusions/Significance

This study provides evidence for the first time in Lepidoptera that females of a moth do not produce any pheromone to attract males, and that mate location is achieved only visually by patrolling males, which may release a pheromone at short distance, putatively a mixture of Z,E-farnesal, E,E-farnesal, and (E,Z)-2,13-octadecadienol. The outlined behavior, long thought to be unique to butterflies, is likely to be widespread in Castniidae implying a novel, unparalleled butterfly-like reproductive behavior in moths. This will also have practical implications in applied entomology since it signifies that the monitoring/control of castniid pests should not be based on the use of female-produced pheromones, as it is usually done in many moths.     

Antennal lobe organization and pheromone usage in bombycid moths

We investigated the neuroanatomy of the macroglomerular complex (MGC), which is involved in sex pheromone processing, in five species in the subfamily Bombycinae, including Ernolatia moorei, Trilocha varians, Rondotia menciana, Bombyx mandarina and Bombyx mori. The glomerulus located at the dorsal-most part of the olfactory centre shows the largest volume in moth species examined to date. Such normal glomerular organization has been observed in E. moorei and T. varians, which use a two-component mixture and includes the compound bombykal as a mating signal. By contrast, the other three species, which use another component as a single attractant, exhibited a modified arrangement of the MGC. This correlation between pheromone usage and neural organization may be useful for understanding the process of speciation.