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In recent years, several invasive woodborers (Coleoptera: Cerambycidae) have been found in Japan. Aromia bungii is a worldwide important pest of fruits and ornamental species of the genus Prunus. It invaded Japan in the early 2010s and now causes heavy damage to stone fruit trees. Anoplophora glabripennis and Apriona swainsoni are destructive pests of street, ornamental and horticultural trees. The first step in intercepting these beetles is to detect their presence early in their infestation, as accurate identification is crucial for their management. Ejected frass is a major sign of infestation and likely holds information on the insect. We focused on chemicals in both larvae and frass, and conducted a GC–MS analysis of these three invasive beetles and the native Anoplophora malasiaca. In all four species, 4 or 5 species-specific hydrocarbons were detected in both larvae and frass. These results indicate that analysis of hydrocarbons in frass could allow definitive detection of invasive wood-boring pests. [ABSTRACT FROM AUTHOR]

Mealybug females release sex pheromones to attract conspecific males for mating. It is critical for mealybug males, which are fragile and short-lived, to respond to the pheromone of their species without time- and energy-consuming cross-attractions to other species. Thus, mealybug pheromone systems are considered to have evolved to be species-specific with unique structures in each species and offer an opportunity to study the diversity of pheromone chemistry that mediates intersexual courtship signals. More than 20 mealybug pheromones are reported to be monoterpenes in general, with only one exception, a hemiterpene alcohol esterified with a medium-chain fatty acid (MCFA), found in the Matsumoto mealybug, Crisicoccus matsumotoi. However, it is unknown whether this is truly exceptional, or if similar compounds are used in other related mealybugs. In this study, we isolated and characterized the pheromone of an allied species, the azalea mealybug C. azaleae. Using gas chromatography–mass spectrometry, nuclear magnetic resonance spectroscopy, and bioassays with synthetics, the pheromone was shown to be composed of isopropyl (E)-7-methyl-4-nonenoate, isopropyl (E)-7-methyl-4-octenoate, and ethyl (E)-7-methyl-4-nonenoate. Surprisingly, the structures of these compounds do not include hemiterpene nor monoterpene motifs but have methyl-branched MCFA parts that are similar to an acid moiety of the C. matsumotoi pheromone. This study implies irregular events for the divergence of pheromone structures in ancestors of the genus Crisicoccus and other mealybugs. [ABSTRACT FROM AUTHOR]

An emerging invader, the red-necked longicorn beetle Aromia bungii (Faldermann) (Coleoptera: Cerambycidae), has become a major pest of cherry blossom, peach, prune, and Japanese apricot in Japan. The synthetic aggregation-sex pheromone, (E)-2-cis-6,7-epoxynonenal, has been used as a chemical for their monitoring. The longevity of the current lure is, however, only one week. Unlike most Lepidopteran lures, it needs to be emitted in large amount of the pheromone emission for effective attraction. In this study, we surveyed chemical retention materials to determine which improving its longevity. The weights of 7 types of lures were measured for 2 or 3 weeks to determine the weight-loss. The pheromone emission from each lure tended to be correlated to the weight-loss. We analyzed the pheromone emissions from candidate lures for three weeks and compared them to the current lures. The current lure continued pheromone emission for 7 d, whereas the candidate lure continued for 14 d. In field trials, the candidate and current lures were both significantly attractive during the first week compared to the control. However, the candidate lure attracted the beetle to the same extent for over 14 d; its longevity was double that of the current lure, therefore, it could be useful for extended monitoring of this beetle. [ABSTRACT FROM AUTHOR]

To evaluate the infection risk of Anoplophora malasiaca (Thomson) (Coleoptera: Cerambycidae) in two species of Japanese pine bonsais (Japanese black pine and Japanese white pine), feeding and oviposition preferences of female A. malasiaca to pine bonsai and mandarin orange were examined. A female was released into a net cage together with a pair of one pine bonsai and one mandarin orange. All mandarin oranges used in preference tests were damaged by feeding and deep biting scars from females, and larval frass ejection from the trunk was found in some mandarin oranges within 4 months of observation. However, these scars were found on only a few pine bonsais, and frass ejection was not found in any pine bonsais. No A. malasiaca larvae were detected in these pine bonsais. Therefore, frass ejection from trunks might be indicative of the presence of larvae. In field surveys at bonsai nurseries, no A. malasiaca adults were found on the pine bonsais and no pine bonsais exhibited frass ejection. Statistical analyses of these results suggested that the risk of A. malasiaca infestation in pine bonsais could be ruled out as a negligible level by confirming an absence from the trunk of frass ejection. [ABSTRACT FROM AUTHOR]

The epiphytic orchid Luisia teres (Asparagales: Orchidaceae) releases floral scent that attracts males of the cupreous polished chafer Protaetia pryeri pryeri (Coleoptera: Scarabaeidae) for pollination. We analyzed this floral scent to identify the attractant(s). When various flower parts were extracted with diethyl ether and assayed, male chafers were attracted to the extract of petals but not to that of sepals or lips. The petal extracts were chromatographed on silica gel column and the activity was found in ether and methanol fractions. Gas chromatography/mass spectrometry (GC/MS) analyses and subsequent NMR analyses revealed that both active fractions contained 2,3-dihydroxypropyl isovalerate (2,3-DHPiV) as a major compound. Natural 2,3-DHPiV showed identical retention time to that of synthetic (R)-enantiomer in subsequent HPLC analyses using a chiral column, while (S)-enantiomer was not detected. The amount of this compound was ca. 5 µg/flower. In field tests performed in Okinawa, Japan, males of P. pryeri pryeri were observed to fly around and land on a cotton ball impregnated with synthetic (R)-2,3-DHPiV, but no male was attracted to the (S)-enantiomer or the racemic mixture. Thus, the chemical attractant released from the flower was identified to be (R)-2,3-DHPiV. 2,3-DHPiV was also found in a volatile sample collected from the virgin females by GC/MS analyses using achiral column, while this compound did not occur in males. Because of the difficulty in HPLC detection of a small amount of 2,3-DHPiV, the chirality remains to be determined. Nevertheless, these results strongly suggest that the flowers of L. teres release the same compound as females of its pollinator P. pryeri pryeri, indicating that the flower of this orchid chemically mimics the female sex pheromone. [ABSTRACT FROM AUTHOR]

The flower bug Orius sauteri is a generalist predator that occurs throughout Japan, and is a promising indigenous natural enemy for micro-pests such as thrips, aphids, and spider mites. We aimed to manipulate the attraction, dispersal, and settlement behavior of Orius bugs using natural chemical substances emitted by the bugs themselves. To identify potential candidates, we screened components in the whole-body extract of O. sauteri based on antennal response and then determined their chemical structure. A gas chromatograph electroantennographic detector (GC/EAD) indicated that the antennae of males responded to two components in the extract of females. GC/mass spectrometry (MS) showed that these two components were octenal and octadienal. Derivatization or GC-FT-IR analysis identified these components as (E)-2-octenal and (E)-2,7-octadienal. To assess the effect of these components on O. sauteri behavior, we conducted two assays. A field bioassay demonstrated that a blend of the two components functioned as a sex pheromone, and a dispersal assay showed that (E)-2-octenal generated a dose-dependent dispersal response. Our study will provide baseline information for enhancing the retention of O. sauteri on important commercial crops to prey on pest species. [ABSTRACT FROM AUTHOR]

The elytra of females of the white-spotted longhorn beetle, Anoplophora malasiaca (Coleoptera: Cerambycidae), are coated with a contact sex pheromone, which was previously shown to be composed of at least three chemical groups. Individually, the chemical groups had little pheromonal activity, but a blend of all three exhibited activity equal to that of the crude female extract. Two groups are female-specific aliphatic hydrocarbons and ketones, which were previously synthesized and confirmed to elicit mating behavior. The third group consists of three lactones, gomadalactones A, B, and C, whose chemical structures were previously identified. These have now been synthesized, and the contact sex pheromone activities of synthetic gomadalactones A, B, and C, and the diastereomer of gomadalactone C, were tested in bioassays in this study. When tested in combination with synthetic hydrocarbons and ketones at the same doses as found in female elytra extract, the individual gomadalactones and a blend showed potent pheromonal activity equivalent to that of the crude extract of the elytra of female beetles. This completes the identification of the essential components of the contact sex pheromone of A. malasiaca. Redundancy of components in the hydrocarbon and ketone groups required to elicit mating behavior was observed previously, and this was also true for the gomadalactones. [ABSTRACT FROM AUTHOR]

Anoplophora malasiaca (Thomson) (Coleoptera: Cerambycidae) is a serious pest that affects various crop trees and landscapes in Japan. We collected and analyzed volatiles from male and female A. malasiaca. 4-(n-Heptyloxy)butan-1-ol and its aldehyde, pheromone components in A. glabripennis and A. chinensis, were detected in the male volatile extracts and nonanal both in the male and female volatile extracts. Nonanal was absent from the extracts of twigs of the willow host plant. Gas chromatograph-electroantennographic responses showed that nonanal and 4-(n-heptyloxy)butan-1-ol elicited responses from both male and female antennae, but 4-(n-heptyloxy)butanal did not. Volatiles of eight artificially reared males, analyzed every 3 or 4 days for 60 days from adult emergence showed that they all produced nonanal and 4-(n-heptyloxy)butan-1-ol. The two compounds produced no short-range female attraction but in males, the short-range attraction to nonanal was dose dependent and significant in a higher dose, but did not depend on 4-(n-heptyloxy)butan-1-ol. When wounded willow twigs were added to nonanal and 4-(n-heptyloxy)butan-1-ol, the frequency of male responses was higher than in all other treatments, but the same as wounded willow twigs alone. The wounded host plant willow twigs might thus be enough for male attraction. The identified volatiles from the beetles in the present study might have a weaker function for attraction from the distance compared to their host plant volatiles. [ABSTRACT FROM AUTHOR]

The red-necked longicorn beetle Aromia bungii (Faldermann) (Coleoptera: Cerambycidae) is an emerging invader in Japan and has become a major pest of cherry blossom trees, peaches, prunes and Japanese apricots. The male-produced aggregation-sex pheromone, recently identified as (E)-2-cis-6,7-epoxynonenal, has two enantiomers. Here, we determined which enantiomer the male produces and tested the responses of beetle antennae to the enantiomers using electroantennography. The male produced only the (6R,7S)-enantiomer, but both enantiomers elicited responses from the antennae of female beetles. Antennae of males also responded to both enantiomers. In field trials, the (6R,7S)-enantiomer and the racemic mixture were equally attractive to both sexes of the beetle, whereas the unnatural (6S,7R)-enantiomer did not attract any beetles. [ABSTRACT FROM AUTHOR]

The red-necked longicorn beetle, Aromia bungii (Faldermann) (Coleoptera: Cerambycidae), which has recently invaded Japan, is a serious pest species. The larvae kill cherry and orchard Rosaceae trees, such as peach, plum, and apricot. To clarify their features for mate location, male and female flight behaviors were observed in wind tunnels with caged males and females as lures. In a small wind tunnel (50 cm in diameter, 2 m in length), both sexes showed takeoff behavior according to increasing airflow. The rate of female takeoff against the male lure, a live male in a wire-netting cage, tended to be higher than for other combinations. In a large wind tunnel (ca. 1.6 m in diameter, 4 m in length), both sexes ascended and showed momentary hovering and astatic flight. When male lure cages were placed windward in the wind tunnel, females showed 'upwind flight,' while males did not. It is assumed that females were induced to fly upwind by a factor derived from live males, likely to be an airborne pheromone component. [ABSTRACT FROM AUTHOR]

The white grub beetle Dasylepida ishigakiensis Niijima et Kinoshita (Coleoptera: Scarabaeidae) is a serious sugarcane pest on the Miyako Islands, Okinawa, Japan. Because this beetle stays underground for most of its lifetime, mating disruption using a synthetic sex pheromone has been suggested as a promising control measure. The amount of pheromone (2-butanol) released by the females is known to decrease drastically as they repeat calling each day. In this study, we determined the response of males to different concentrations of ( R)-2-butanol, both in the laboratory and in the field. Males showed typical pre-mating behaviors and they were attracted to ( R)-2-butanol even at a concentration 1/100 of the amount typically emitted by a female during the 1st period of calling. We examined whether female attractiveness to males was reduced with the frequency of calling, by counting the number of males attracted to females that had called during zero to five periods beforehand. Our results indicated that a high level of female attractiveness was maintained even after three periods of calling. Based on these findings, we propose that releasing synthetic sex pheromone shortly before the start of the mating season and into the late mating season may be optimal for disrupting the mating of this beetle. [ABSTRACT FROM AUTHOR]

The effect of mating disruption by the ground-surface application of 2-cm-long dispensers (mini-dispensers) of 2-butanol against the white grub beetle Dasylepida ishigakiensis Niijima et Kinoshita (Coleoptera: Scarabaeidae) was examined in sugarcane fields on the Miyako Island, Okinawa, Japan. Mating rates and male catches with sex pheromone traps were reduced to a low level comparable to that obtained from a conventional method in which rope dispensers (25 m) were hung at a height of ca. 30 cm along the sugarcane ridges. Both mating rates and male catches were reduced with increasing number of treated mini-dispensers. These results suggest that by using mini-dispensers the amounts of synthetic sex pheromone and plastic resin can be reduced to 1/10-1/5 and 1/6-1/3, respectively, of their conventional application with rope-type dispensers, without impairing the efficiency of mating disruption in this beetle. Furthermore, the amount of labor required for the application of this method is expected to be greatly reduced compared to the rope-type dispenser method. [ABSTRACT FROM AUTHOR]

Orius minutus (L.) (Heteroptera: Anthocoridae) is a natural enemy of agricultural pests such as thrips, aphids, and various newly hatched insect juveniles. In this study, we conducted 1) behavioral assays for evidence of contact sex pheromone activity in trails of O. minutus, and 2) chemical analysis to identify the essential chemical components of the trails. Males showed arrestment to trails of mature virgin females but not to trails from either conspecific nymphs or immature females. Females also showed arrestment to trails from conspecific males, although the response was weaker than that exhibited by males. The activity of female trails lasted for at least 46 h after deposition. Males showed a response irrespective of mating experience. Following confirmation that a contact sex pheromone was present in the trails of female O. minutus, we used a bioassay-driven approach to isolate the active chemicals. After fractionation on silica gel, the n-hexane fraction was found to be biologically active to males. A major compound in the active fraction was ( Z)-9-nonacosene; this compound was found only in trail extracts of mature virgin females. Synthetic ( Z)-9-nonacosene arrested O. minutus males, indicating that it is the major active component of the contact sex pheromone in the trails of female O. minutus. [ABSTRACT FROM AUTHOR]

The use of pheromones to disrupt mating behaviours is perhaps the most promising method for controlling the sugarcane pest Dasylepida ishigakiensis Niijima et Kinoshita, also known as the white grub beetle. Herein, based on laboratory observations of mating, oviposition and hatching behaviours in the white grub beetle, we suggest guidelines for the timing and length of sex-pheromone release in the field. In the present study, we collected virgin female adults early in the mating season (4 February) in sugarcane fields in Okinawa, Japan and mated them in the laboratory after different time intervals with males collected at the same time. We investigated the effects of delayed mating on mating success, timing of oviposition, female longevity after mating and progeny hatchability. Mating success – defined as the proportion of mated females that produced progeny – and hatchability were not significantly affected by mating delays of less than 31 days, although further delays dramatically decreased these parameters. Female life span and the time between mating and oviposition were unaffected by mating delay. Similar results were obtained for laboratory-reared adults mated on day 45, 60 or 80 after adult emergence. Based on these findings, we propose that releasing synthetic sex pheromones between mid-January (i.e. shortly before the start of the mating season) and late March would be optimal for disrupting the mating behaviours of this beetle in the field. [ABSTRACT FROM PUBLISHER]

The feasibility of mating disruption with synthetic sex pheromone for the control of the white grub beetle Dasylepida ishigakiensis (Coleoptera: Scarabaeidae) was examined by permeating sugarcane fields with a racemic mixture of 2-butanol ( rac-2B) released from polyethylene-tube dispensers on Miyako Island, Japan in 2011. An application of rac-2B released from 10,000 tube dispensers into 3,200 m sugarcane fields significantly reduced the mating rate of feral females and catches of feral males with R2B-baited traps compared with the results from untreated sugarcane fields. The larval density for the treated fields was found to be nearly zero in the following winter when the corresponding figure for untreated fields was high (1.73 and 2.33/40 × 40 cm quadrat). These results clearly show that the mating disruption technique using rac-2B could be highly promising for the control of D. ishigakiensis in sugarcane fields. [ABSTRACT FROM AUTHOR]

The use of magnetic resonance imaging (MRI) as a tool for in vivo detection of eggs in living Dasylepida ishigakiensis Niijima et Kinoshita, a major pest of sugarcane, was explored using females with an ovary at different developmental stages. MRI measurements of beetles were performed at 13 °C to avoid motion artifacts on the MR images. Spin-lattice relaxation time-weighted images allowed the observation of eggs at short acquisition times (2 min, 8 s). By comparing MR images with dissection data, criteria for determining mature eggs in MR images were a clear circular or ellipsoidal shape surrounded by a relatively bright rim and a size typically larger than 1.3 mm in the minor axis. Although small oocytes could not be detected, females with a developed or undeveloped ovary could be clearly distinguished based on MR images. The possibility of confusing the digestive tract as eggs in a female with a less developed ovary can be eliminated using a proton density weighted image. [ABSTRACT FROM AUTHOR]

The subtropical scarab beetle, Dasylepida ishigakiensis, has a two-year life cycle. This study showed the time of adult emergence from the soil relative to the time of dusk and the presence of the female sex pheromone. Beetles collected on Miyako Island were transported to Tsukuba where they were immediately placed under natural day lengths in February. They exhibited two emergence peaks that corresponded to the times of dusk in Tsukuba and on the island, respectively. Males emerged precociously if a lure containing synthetic female sex pheromone was placed in their container, whereas the females' behavior was unaffected. Previous observations that mated females dig deeper in the soil than virgin females, males or mated males were confirmed. To explore the underlying mechanism controlling the behavioral change associated with mating, liquid material derived from the male accessory glands, seminal vesicles and female bursa copulatrix was injected into beetles, but without any significant influence on burrowing behavior. No significant influence was also observed in beetles injected with anisomycin, an inhibitor of protein synthesis related to memory in other animals. [ABSTRACT FROM AUTHOR]

Adult Anoplophora malasiaca beetles were collected from field populations inhabiting three different host plants: Citrus, Salix and Vaccinium. Male orientation responses towards odours of wounded branches of each host plant were examined. Males of all three populations were attracted most frequently to odours of their original hosts, indicating that male orientation behaviour was induced by the odours of the plants from which they were originally collected. We also observed the mating responses of the males to elytra extracts from young and mature females of the three populations and chemically analysed those extracts. Citrus males were highly responsive to female extracts, regardless of origin. Salix males were less reactive than Citrus males, but responded moderately to female extracts of all populations. Vaccinium males were never attracted to Citrus female extracts and responded more to extracts from mature Vaccinium and Salix females, indicating that males might distinguish female origin and maturity. All eight hydrocarbons, four ketones, and three lactones that were previously identified as female contact sex pheromones were found in the extracts of three populations and both ages. Higher amounts of lactones on mature females likely caused greater male responses. These results suggested that males would be more likely to approach a female feeding on the same host plant and subsequently recognise her sexual maturity based on the profile of contact sex pheromones on her elytra. [ABSTRACT FROM AUTHOR]

The effects of changes in host plants on the mate-searching behavior and feeding preferences of the white-spotted longicorn beetle Anoplophora malasiaca (Thomson) (Coleoptera: Cerambycidae) were examined. All individuals were raised on the same artificial diet until they became pupae. Analysis of the mate-searching behavior of the males showed that many more newly emerged males were attracted to the odor of the artificial diet than to an unbaited control. We prepared three groups of beetles and fed each group on different host plants for one week. The host plants used included the following: an artificial diet (containing Morus alba Linné), Citrus unshiu Marc. branches, and Vaccinium spp. branches. The mate-searching behavior of the males changed in relation to the plant supplied for feeding. Simultaneously, the preference among the three host plants was tested. The newly emerged males preferred the artificial diet. After a week of feeding on one of the three plants, however, the adult males selected and consumed significantly more of the plant that they had just experienced than the other plants. These results suggest that the male mate-location cue can be acquired after adult eclosion. In addition, the male beetles are capable of changing their host-plant preference. If they do so, they use different odor cues for mate location. Newly emerged A. malasiaca females showed no preference for their first choice of food among the three host plants presented, whereas the consumption was significantly larger on C. unshiu branches. After one week of feeding on different host plants, females chose their host plant after the adult stage as well as C. unshiu, but consumed mostly C. unshiu. These results suggest that the food preferences of females are different from those of males. The behavior of females may not be affected by chemical signals from their original host-plant species (as pupae) or from the host-plant species acquired when they emerge as adults. [ABSTRACT FROM AUTHOR]

Males of the white grub beetle, Dasylepida ishigakiensis Niijima et Kinoshita, transfer a large amount of a colloidal substance to females during mating. In this study we investigated the effect of the male substance on the reproductive performance of mated females and the significance of multiple mating of this beetle. Females artificially separated from the males 5 min after the start of mating produced fewer eggs than those separated after 30 min or those that were undisturbed and separated spontaneously, suggesting that the male substance is used as a nutrient for egg production by the females. When females were allowed to mate with 1-4 males, multiple mating had no clear effect on reproductive performance. The amount of male substance stored in the bursa copulatrix (BC) was not significantly increased by a second mating. The functions of multiple mating of this species may be to provide a chance for females to obtain sufficient amounts of male substance when the first male to mate has only small amounts of this substance, and to increase the genetic heterogeneity of the progeny. The presence of a serine proteinase and its possible involvement in the dynamics of the BC contents are reported. [ABSTRACT FROM AUTHOR]

In order to understand the visual orientation mechanisms of the male black chafer Holotrichia loochooana loochooana to females, we examined male behavioral response to colored objects (lures) when female sex pheromone was present. In the field, males landed more frequently on darker lures than on lighter ones of the same color. Among lures of different colors but with the same brightness, males preferred landing on black, blue, or red ones to green or grey ones. The reflectance intensities of preferred lures were weaker than those of green and grey in the range of 540-580 nm. This range is considered to affect choice by males. When females perch on the leaf edges of the food plant to adopt a calling posture, the dominant background color is that of the leaves (i.e., green). This means that the landing target forms a silhouette (dark spot) which is prominent against a background of green leaves, providing maximum visual acuity for mating. [ABSTRACT FROM AUTHOR]

Volatile chemicals from Citrus and Salix host plants evoke orientation behavior in males of the species Anoplophora malasiaca (Thomson) (Coleoptera: Cerambycidae). These chemicals are emitted from wounded branches. We hypothesized that when released, these chemicals may indicate the presence of an individual to other conspecifics. Insects that originate from different host plants may use different plant chemicals from their own host to communicate with conspecifics. To further explore this theory, we investigated this communication system in a population of A. malasiaca from a third host plant, blueberry ( Vaccinium spp.). Males from a blueberry host ( Vaccinium population) were attracted to the odor of wounded Vaccinium branches when released near a female model in the laboratory, as has been observed in males found on Citrus and Salix host plants. The Vaccinium branch extract that was attractive to the males was separated into six fractions, of which two were active. Three active compounds were subsequently identified: β-caryophyllene and sulfur from the hexane fraction, and ( E)-phytol in the weakly polar fraction. The latter two active compounds of Vaccinium branches were different from those found in Citrus and Salix. [ABSTRACT FROM AUTHOR]

In 2010, abrupt outbreaks of the African armyworm, Spodoptera exempta (Walker), occured on the Tarama, Iriomote and Kikai Islands in southwestern Japan. Analysis by gas chromatography-electroantennographic detection (GC-EAD) revealed two EAG-active compounds on male antenna in crude extract of virgin females. These compounds were identified as (9 Z)-9-tetradecenyl acetate (Z9-14:Ac) and (9 Z,12 E)-9,12-tetradecadienyl acetate (Z9E12-14:Ac) in ca. 90:10 ratio by subsequent GC-MS analyses. (11 Z)-11-Hexadecenyl acetate (Z11-16:Ac), which had previously been identified as a third component in the Kenyan population, was not detected. Binary blends of Z9-14:Ac and Z9E12-14:Ac at ratios between 99:1 and 90:10 showed a potent attractiveness in the field, superior to that of virgin females and comparable to that of the three-component formulation determined in Kenya. For the population survey, a 98:2 blend was used. In Tarama, only a few moths of S. exempta were captured with a light trap during the night when more than 600 males were captured with synthetic sex pheromone; more S. exempta captures with a light trap had been reported than with sex-pheromone traps in Kenya. This indicates that the Okinawan population has different properties from the Kenyan population in pheromone composition and behavioral response to light. [ABSTRACT FROM AUTHOR]

Genetic variation of the white-spotted longicorn beetle Anoplophora spp., distributed on the Japanese mainland and the Ryukyu Islands, was examined using nucleotide sequences of mitochondrial DNA. Two fragments, a 1.2-kb-long fragment containing portions of the cytochrome oxidase subunit I and II genes and a 1.4-kb-long fragment containing portions of 16S and 12S rDNAs, were sequenced. In phylogenetic analyses based on the sequences, 294 individuals collected at 75 localities were divided into two major groups (groups A and B) and then split into seven subgroups (A1-A4 and B1-B3). In group A, the closely related subgroups A1, A2, and A3 were mainly distributed on the Japanese mainland and were roughly separated among geographic areas, although the range of A3 spread significantly from Kyushu to the Central Ryukyu Islands. The sequence of A4, detected from one individual collected in eastern Honshu, was almost the same as that reported for A. chinensis (Forster) on the Chinese continent. In group B, subgroups B1 and B2 were restricted to the Central and Southern Ryukyu Islands, respectively, while B3 was distributed widely in both regions. Based on the results, we discuss the geographic origin of the haplotypes and movement of the insects between geographic areas. [ABSTRACT FROM AUTHOR]

Male white-spotted longicorn beetles Anoplophora malasiaca raised on willow Salix schwerinii were preferentially attracted to the odour of wounded S. schwerinii branches when released near a female model in the laboratory. This attractiveness rapidly decreased within 2 h after wounding. Solid phase microextraction and subsequent gas chromatography analyses of volatiles from the wounded branches detected three highly volatile monoterpenes: 1,8-cineole, nerol and geraniol. Among the monoterpene compounds, nerol showed a significant attractiveness to the male beetles raised on Salix. Sesquiterpene hydrocarbons, which have been identified as attractants for a beetle population raised on citrus, were detected only in trace amounts in S. schwerinii branches. These results suggest that beetles raised on these two hosts use different odour cues for mate location. [ABSTRACT FROM AUTHOR]

Field and laboratory observations were carried out to understand the significance of prolonged copulation and the male substance transferred from males to females in the white grub beetle Dasylepida ishigakiensis Niijima & Kinoshita. Adults emerged from the soil in the evening and formed mating pairs on sugarcane plants, where copulation lasted for 1–3 h. Mating pairs (30%) observed in the field were accompanied by one or two additional males. Some females mated with one such male after ending copulation with the first male, showing multiple mating. After mating, males dropped to the ground to burrow into the soil earlier than females. Females received a large amount of colloidal secretion from males during mating. The amount and rate of transfer of this secretion from males to females were investigated in this study. Transfer of the secretion from males to females was completed within 30 min, and females that had mated with two males in the laboratory stored a larger amount of male secretion in the bursa copulatrix than those that had mated once. The results of both field and laboratory observations might indicate a possibility that prolonged copulation serves as a post-insemination female guarding behaviour to prevent mating by other conspecific males. [ABSTRACT FROM PUBLISHER]

The privet tree, Ligustrum obtusifolium (Oleaceae), defends its leaves against insects with a strong lysine-decreasing activity that make proteins non-nutritive. This is caused by oleuropein, an iridoid glycoside. We previously found that some privet-specialist caterpillars adapt by secreting glycine in the digestive juice as a neutralizer that prevents the loss of lysine. Here, we extended the survey into 42 lepidopteran and hymenopteran species. The average concentration of glycine in digestive juice for 11 privet-feeding species (40.396 mM) was higher than that for 32 non-privet-feeding species (2.198 mM). The glycine concentrations exceeded 10 mM in 7 out of 11 privet-feeding species. In Macrophya timida (Hymenoptera), it reached 164.8 mM. Three out of the four remaining privet-feeding species had other amino acids instead. Larvae of a privet-specialist butterfly, Artopoetes pryeri (Lycaenidae), had a high concentration (60.812 mM) of GABA. In two other specialists, β-alanine was found. GABA, β-alanine, and glycine as well as alanine, amines, and ammonium ion inhibited the lysine decrease, indicating that amino residues are responsible for the inhibition. However, the three amino acids found in the specialists were far more effective (20 mM showed 80% inhibition) than the rest (>140 mM was required for 80% inhibition). Our results show a clear and rare case of the apparent convergent evolution of herbivores' molecular adaptations of feeding on a plant with a chemical defense in a manner that minimizes the cost of adaptation. The novel role of GABA in plant-herbivore interactions shown here is probably the first reported non-neuronal role of animal-derived GABA. [ABSTRACT FROM AUTHOR]

A predatory stink bug, Eocanthecona furcellata, died after feeding on Milionia basalis larvae. The compounds toxic to E. furcellata were isolated from the hemolymph of M. basalis larvae and identified as inumakilactone A, nagilactone C, and nagilactone C glucoside. The concentrations of inumakilactone A, nagilactone C, and nagilactone C glucoside in the hemolymph of the final instar larvae were 130, 50, and 770 μg/ml, respectively. Nagilactone C showed the highest insecticidal activity against second-instar nymphs of E. furcellata, while nagilactone C glucoside showed the lowest, one twentieth of that of nagilactone C. When mixed compounds were given at the same concentrations as those in hemolymph of M. basalis, all nymphs of E. furcellata died with in three days. Inumakilactone A and nagilactone C were found to be in the leaves of podocarp, Podocarpus macrophyllus, the only host plant of M. basalis, at concentrations of 13 and 175 μg/g fresh weight, respectively. However, no nagilactone C glucoside was detected in the leaves of this species. These results suggested that M. basalis may transform nagilactone C to its glucoside. [ABSTRACT FROM AUTHOR]

Bitter gourd, Momordica charantia, was less palatable to two species of armyworms, Spodoptera litura and Pseudaletia separata, than two other cucurbitaceous plants. A methanol extract of M. charantia leaves inhibited feeding of the armyworm larvae. The two most active fractions obtained by silica gel chromatography were purified by HPLC. Momordicine II, a triterpene monoglucoside, was identified as an antifeedant compound from the more active of these fractions. The second active fraction led to the isolation of a new triterpene diglucoside. Fresh leaves of M. charantia contained ca. 0.3% of momordicine II. Momordicine II showed significant antifeedant effects on P. separata at concentrations of 0.02, 0.1, and 0.5% in artificial diets. Momordicine II caused a significant feeding reduction in S. litura only at the highest concentration (0.5%) tested. The difference in feeding response of the two armyworms to momordicine II may be related to the diversity in their host range. [ABSTRACT FROM AUTHOR]

In previous studies, we reported the existence of a high concentration of free glycine in the digestive juice of several Lepidoptera larvae, particularly in the digestive juice of species that feed on the privet tree, Ligustrum oblusifolium. The water extract of privet leaves showed very strong protein-denaturing activity and lysine-decreasing activity, which closely resembled activity of oxidized polyphenolics. Addition of 1% glycine to the extract could completely inhibit these activities. Free glycine may be secreted into the digestive juice by larvae as an adaptive mechanism for chemical defense against its host plants. The protein-denaturing compound in privet leaves is present in the cytosol or in the vacuoles of the leaf cells. The compound does not show protein-denaturing activity without oxidation, but when mixed with intact organelles under low osmotic conditions to give an osmotic shock, a very high protein-denaturing activity is produced. Our results suggest that the privet tree is endowed with a defense mechanism in which a stable compound in the cytosol or in the vacuoles is activated into a chemically active denaturant by an enzyme present in the organelles (including chloroplasts) after the leaves are eaten by insects and the organelles are broken by osmotic shock or by digestive mechanisms. Based upon HPLC and NMR data, we conclude that the denaturing compound is oleuropein, an o-dihydroxyphenolic compound. This compound makes up 3% of the wet weight of privet leaves. The protein-denaturing activity of purified oleuropein activated by the leaf enzyme is high enough to account for all the denaturing activity in a water extract of privet leaves. The denaturing reaction is completely inhibited by free glycine. Our results suggest that the protein-denaturing activity and lysine-decreasing activity of privet leaves are caused by oxidized polyphenolics, and that some insects secrete free glycine to counter the denaturing activity of oxidized phenolics. The chemical mechanism of counteraction by glycine is also considered. Free glycine in the midgut of insects probably protects proteins from denaturation by competing with proteins for oxidized phenolics. [ABSTRACT FROM AUTHOR]