Silkworms suppress the release of green leaf volatiles by mulberry leaves with an enzyme from their spinnerets.

In response to herbivory, plants emit a blend of volatile organic compounds that includes green leaf volatiles (GLVs) and terpenoids. These volatiles are known to attract natural enemies of herbivores and are therefore considered to function as an indirect defense. Selection should favor herbivores that are able to suppress these volatile emissions, and thereby make themselves less conspicuous to natural enemies. We tested this possibility for silkworms, which were observed to leave secretions from their spinnerets while feeding on mulberry leaves. When we ablated the spinnerets of silkworms, no secretions were observed. Leaves infested by intact silkworms released smaller amounts of GLVs than leaves infested by ablated silkworms, indicating that the spinneret secretion suppressed GLV production. This difference in GLV emissions was also reflected in the behavioral response of Zenillia dolosa (Tachinidae), a parasitoid fly of silkworms. The flies laid fewer eggs when exposed to the volatiles from intact silkworm-infested leaves than when exposed to the volatiles from ablated silkworm-infested leaves. We identified a novel enzyme in the secretion from the spinneret that is responsible for the GLV suppression. The enzyme converted 13(S)-hydroperoxy-(9Z,11E,15Z)-octadecatrienoic acid, an intermediate in the biosynthetic pathway of GLVs, into its keto-derivative in a stereospecific manner. Taken together, this study shows that silkworms are able to feed on mulberry in a stealthy manner by suppressing GLV production with an enzyme in secretions of their spinnerets, which might be a countermeasure against induced indirect defense by mulberry plants.

(1S,3R)-cis-Chrysanthemyl Tiglate: Sex Pheromone of the Striped Mealybug, Ferrisia virgata.

Derivatives of 2,2-dimethyl-3-(2-methylprop-1-enyl)cyclopropanecarboxylic acid (chrysanthemic acid) are classic natural pyrethroids discovered in pyrethrum plants and show insecticidal activity. Chrysanthemic acid, with two asymmetric carbons, has four possible stereoisomers, and most natural pyrethroids have the (1R,3R)-trans configuration. Interestingly, chrysanthemic acid-related structures are also found in insect sex pheromones; carboxylic esters of (1R,3R)-trans-(2,2-dimethyl-3-(2-methylprop-1-enyl)cyclopropyl)methanol (chrysanthemyl alcohol) have been reported from two mealybug species. In the present study, another ester of chrysanthemyl alcohol was discovered from the striped mealybug, Ferrisia virgata (Cockerell), as its pheromone. By means of gas chromatography-mass spectrometry, nuclear magnetic resonance spectrometry, and high-performance liquid chromatography analyses using a chiral stationary phase column and authentic standards, the pheromone was identified as (1S,3R)-(-)-cis-chrysanthemyl tiglate. The (1S,3R)-enantiomer strongly attracted adult males in a greenhouse trapping bioassay, whereas the other enantiomers showed only weak activity. The cis configuration of the chrysanthemic acid-related structure appears to be relatively scarce in nature, and this is the first example reported from arthropods.

Sex pheromone of a coccoid insect with sexual and asexual lineages: fate of an ancestrally essential sexual signal in parthenogenetic females.

Sex pheromones play a central role in intersexual communication for reproduction in many organisms. Particularly in insects, reproductive isolation that leads to speciation is often achieved by shifts of pheromone chemistries. However, the divergence and evolution of pheromones remain largely unknown. This study reveals a unique evolutionary consequence for terpenoid pheromones in coccoid insects. Coccoids, such as mealybugs, show clear sexual dimorphism: males are dwarf and short-lived, whereas females are wingless and almost immobile. Female pheromones are therefore indispensable for males to navigate for sexual reproduction, but some females can reproduce asexually. Interestingly, a derived asexual lineage that reproduces by parthenogenesis coexists with its ancestral lineage that reproduces sexually in a population of the pineapple mealybug, Dysmicoccus brevipes Here, we isolated, characterized and synthesized a novel monoterpene, (-)-(anti-1,2-dimethyl-3-methylenecyclopentyl)acetaldehyde, as a pheromone of the sexual females of Dbrevipes This monoterpene aldehyde, with an irregular linkage of isoprene units, is notable, because all mealybug pheromones previously reported are carboxylic esters of terpenols. This compound was, however, never produced by the asexual females. As a consequence of acquiring parthenogenetic reproduction, the asexual females appear to have abandoned the production of the sex pheromone, which had been essential to attracting males in their ancestors.

Sex Pheromone of the Cotton Mealybug, Phenacoccus solenopsis, with an Unusual Cyclobutane Structure.

The cotton mealybug, Phenacoccus solenopsis, the distribution of which was formerly limited to Nearctic and Neotropical regions, recently invaded many countries in various regions including Asia, Africa, and the Pacific. More recently, P. solenopsis was newly recorded in Japan and is currently an emerging pest of agricultural crops. In this study, we determined the structure of a sex pheromone of P. solenopsis in order to develop an effective lure for monitoring this pest. From volatiles emitted by virgin adult females, we isolated a compound attractive to males. By means of coupled gas chromatography-mass spectrometry and nuclear magnetic resonance spectroscopy, we identified this as (2,2-dimethyl-3-isopropylidenecyclobutyl)methyl 3-methylbut-2-enoate. This compound was synthesized and shown to be attractive to male P. solenopsis. Analysis by gas chromatography using an enantioselective stationary phase and polarimetry analyses of the natural pheromone and synthetic enantiomers showed the natural compound to be the (R)-(-)-enantiomer. This compound is an ester of maconelliol, which has an unusual cyclobutane structure found in sex pheromones of other mealybug species, and senecioic acid, also found in the pheromones of other mealybug species. However, this is the first example of the ester of maconelliol and senecioic acid as a natural product.

Sexual versus Asexual Reproduction: Distinct Outcomes in Relative Abundance of Parthenogenetic Mealybugs following Recent Colonization.

Asexual reproduction, including parthenogenesis in which embryos develop within a female without fertilization, is assumed to confer advantages over sexual reproduction, which includes a "cost of males." Sexual reproduction largely predominates in animals, however, indicating that this cost is outweighed by the genetic and/or ecological benefits of sexuality, including the acquisition of advantageous mutations occurring in different individuals and the elimination of deleterious mutations. But the evolution of sexual reproduction remains unclear, because we have limited examples that demonstrate the relative success of sexual lineages in the face of competition from asexual lineages in the same environment. Here we investigated a sympatric occurrence of sexual and asexual reproduction in the pineapple mealybug, Dysmicoccus brevipes. This pest invaded southwestern Japan, including Okinawa and Ishigaki Islands, in the 1930s in association with imported pineapple plants. Our recent censuses demonstrated that on Okinawa sexually reproducing individuals can coexist with and even dominate asexual individuals in the presence of habitat and resource competition, which is considered to be severe for this nearly immobile insect. Molecular phylogeny based on partial DNA sequences in the mitochondrial and nuclear genomes, as well as the endosymbiotic bacterial genome, revealed that the asexual lineage diverged from a common sexual ancestor in the relatively recent past. In contrast, only the asexual lineage exhibiting obligate apomictic thelytoky was discovered on Ishigaki. Co-existence of the two lineages cannot be explained by the results of laboratory experiments, which showed that the intrinsic rate of increase in the sexual lineage was not obviously superior to that of the asexual lineage. Differences in biotic and/or abiotic selective forces operating on the two islands might be the cause of this discrepancy. This biological system offers a unique opportunity to assess the relative success of sexual versus asexual lineages with an unusual morphology and life cycle.

A new lavandulol-related monoterpene in the sex pheromone of the grey pineapple mealybug, Dysmicoccus neobrevipes.

The grey pineapple mealybug, Dysmicoccus neobrevipes, is a serious pest that attacks a variety of crops in tropical regions. Recently, it was recorded on an island in southwestern Japan, suggesting that its distribution is expanding. As a measure against this expansion, a monitoring tool is urgently needed. In this study we determined the structure of the sex pheromone of D. neobrevipes in order to develop an efficient lure for monitoring traps. Volatiles collected from virgin adult females were fractionated by high performance liquid chromatography (HPLC) and gas chromatography, and fractions were tested for attractiveness to males in a laboratory bioassay. A single compound was isolated which was as attractive to males as the crude collections, and this was proposed to be the main, if not the only, component of the female-produced sex pheromone. The structure of this was determined to be (E)-2-isopropyl-5-methylhexa-3,5-dienyl acetate by gas chromatography/mass spectrometry and nuclear magnetic resonance analyses. This compound was synthesized through four steps, and the synthetic chemical was as attractive as the natural product in a greenhouse bioassay. The enantiomers of the synthetic acetate were obtained by enantioselective HPLC fractionation of the corresponding alcohols, and the natural pheromone was shown to be the (+)-isomer. The carbon skeleton of this novel compound is related to lavandulol, a monoterpene with an unusual non-head-to-tail connection of isoprene units that is often found in mealybug pheromones.

Different uses of plant semiochemicals in host location strategies of the two tachinid parasitoids.

Some members of the family Tachinidae (Insecta: Diptera) deposit numerous very small eggs, termed "microtype" eggs, on the food plants of their caterpillar hosts. Parasitization is successful only when the hosts ingest these eggs. To increase the chance of hosts encountering the eggs, microtype tachinid parasitoids have to choose a suitable plant that harbors hosts and lay their eggs near the hosts. In their host location process, semiochemicals emitted by host-infested plants offer the tachinids a reliable cue. We investigated the behavioral responses of two microtype tachinid parasitoids, Pales pavida and Zenillia dolosa, to maize plants infested with their caterpillar host, Mythimna separata, in a wind tunnel. P. pavida females showed a significantly higher rate of landing on caterpillar-infested plants than on mechanically wounded or intact plants, whereas Z. dolosa landed on both the caterpillar-infested and mechanically wounded plants at significantly higher rates than on intact plants. We also examined which part of a caterpillar-infested maize leaf induces oviposition. P. pavida deposited eggs on the margin of the leaf, whereas Z. dolosa preferentially laid eggs around a caterpillar-infested area or a mechanically wounded spot. P. pavida eggs retained their parasitization ability for more than 15 days after they were deposited, whereas the eggs of Z. dolosa could not survive more than 5 days after oviposition. Our results suggest that each tachinid parasitoid employs a different host location strategy to exploit semiochemicals coming from plant-herbivore interaction as cues in order to increase their parasitization success.

A comparison of artificial diets for mass rearing Brontispa longissima (Coleoptera: Chrysomelidae) as hosts for the larval and the pupal parasitoids.

The coconut hispine beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae) is a serious pest of coconut palm. In this study, we developed an artificial diet for B. longissima so that the beetle could be used as a host for rearing two of its parasitoids, Asecodes hispinarum Boucek (Hymenoptera: Eulophidae) and Tetrastichus brontispae Ferrière (Hymenoptera: Eulophidae). The new artificial diet represents an improvement of our previous diet, which we used as a control. When beetle larvae were reared on the new diet, which contains cysteine but not cellulose powder and has twice as much coconut leaf powder as in the control, the adult emergence was 71% (approximately 2 times that in the control). We also examined the suitability of beetles fed on the new diet as hosts for the larval parasitoid A. hispinarum and the pupal parasitoid T. brontispae. The percentage of wasps that emerged from hosts that were fed the new diet was higher than that from the control-fed hosts. The new diet allowed both A. hispinarum and T. brontispae to produce adult wasps of the next generation, whereas the control only allowed T. brontispae to produce the next generation. These results suggest that the new diet is suitable for B. longissima and will facilitate mass-rearing of A. hispinarum and T. brontispae.

Attraction to herbivore-induced plant volatiles by the host-foraging parasitoid fly Exorista japonica.

Responses of the tachinid fly Exorista japonica Townsend to odors from corn plants infested with the fly's host, the larvae of the noctuid moth Mythimna separata (Walker), were examined in a wind tunnel. Naïve female flies showed a higher rate of landing on M. separata-infested corn plants from which the host larvae had been removed than on artificially damaged or intact corn plants. When paper impregnated with a solution of headspace volatiles collected from host-infested plants was attached to intact plants, females landed on the plants at a high rate. Females also responded to intact plants to which had been attached with paper impregnated with a synthetic blend of nine chemicals identified previously in host-infested plants. There was an optimum concentration of the synthetic blend for the females' landing. Of the nine chemicals identified previously, four [(E)-4,8-dimethyl-1,3,7-nonatriene, indole, 3-hydroxy-2-butanone, and 2-methyl-1-propanol] released only by host-infested plants were classified as a host-induced blend. The other five [(Z)-3-hexen-1-yl acetate, (E)-2-hexenal, hexanal, (Z)-3-hexen-1-ol, and linalool] were classified as a non-specific blend released not only by infested plants but also by artificially damaged or intact plants. In the wind tunnel, E. japonica females did not respond to intact plants to which paper containing a solution of non-specific blend or host-induced blend was attached. However, they showed a high level of response to a mixture of the non-specific and host-induced blends. These results indicate that naïve E. japonica use a combination of non-specific and host-induced blends as an olfactory cue for locating host-infested plants.