Nao Fujiwara-Tsujii

Volatile attractant phytochemicals for a population of white-spotted longicorn beetles Anoplophora malasiaca (Thomson) (Coleoptera: Cerambycidae) fed on willow differ from attractants for a population fed on citrus

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 2xa0h 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.

Determination of the amount of sex pheromone emitted by individual virgin females of the white grub beetle, Dasylepida ishigakiensis (Coleoptera:Scarabaeidae) at different stages of reproductive life

Virgin females of the white grub beetle, Dasylepida ishigakiensis Niijima et Kinoshita, emit the sex pheromone, 2-butanol. To investigate the relationships between pheromone emission and ovarian development in this beetle, virgin adult females were kept at 18°C for various lengths of time. The sex pheromone released by individual females at first calling was collected at 23°C and the amounts determined. Females started developing visible oocytes at dayxa044 and emitting sex pheromone at dayxa050. Almost all females at dayxa052 or later possessed eggs, indicating that diapause had ended. A high positive correlation was found between the amounts of pheromone emitted and levels of ovarian development. A large variation of pheromone emission was encountered even among individuals of the same age. Females retained the ability to emit sex pheromone as long as they were virgin. These results may provide important information regarding the timing and duration of application of synthetic sex pheromone in an attempt to control this pest through disrupting the male-female communication.

Population differences in male responses to chemical mating cues in the white-spotted longicorn beetle, Anoplophora malasiaca

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.

Mating disruption for control of the white grub beetle Dasylepida ishigakiensis (Coleoptera: Scarabaeidae) with synthetic sex pheromone in sugarcane fields.

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 3,200xa0m2 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 2.33/40xa0×xa040xa0cm 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.

The white-spotted longicorn beetle, Anoplophora malasiaca (Coleoptera: Cerambycidae), with a blueberry as host plant, utilizes host chemicals for male orientation

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.

The significance of multiple mating and male substance transferred to females at mating in the white grub beetle, Dasylepida ishigakiensis (Coleoptera: Scarabaeidae)

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 5xa0min after the start of mating produced fewer eggs than those separated after 30xa0min 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.

Sex pheromone of the African armyworm Spodoptera exempta (Lepidoptera: Noctuidae): identification of components of the Okinawan population and formulation for population monitoring

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 (9Z)-9-tetradecenyl acetate (Z9-14:Ac) and (9Z,12E)-9,12-tetradecadienyl acetate (Z9E12-14:Ac) in ca. 90:10 ratio by subsequent GC-MS analyses. (11Z)-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.

Extrinsic and Intrinsic Factors Influencing the Emerging and Burrowing Behaviors of Reproductively Active Adults of the White Grub Beetle, Dasylepida ishigakiensis

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.

Managing the white grub beetle Dasylepida ishigakiensis (Coleoptera: Scarabaeidae) in sugarcane fields on Miyako Island, Japan, using sex attractant pheromone: effects of mating delay on the reproductive ability of laboratory-reared and field-collected females

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.

The effects of foods consumed after adult eclosion on the mate-searching behavior and feeding preferences of the white-spotted longicorn beetle Anoplophora malasiaca (Coleoptera: Cerambycidae)

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 onexa0week. 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 onexa0week 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.