Male white grub beetles prefer the pheromone composition of young females in the field

Females of the white grub beetle, Dasylepida ishigakiensis, release both (R)- and (S)-2-butanol as sex pheromones, but the males are only attracted to (R)-2-butanol. In laboratory-reared females, the proportion of the (R)-isomer decreased significantly as their calling opportunities increased and as they aged. We examined whether such qualitative changes also occur in field populations. We collected virgin females from the field and then trapped and analysed the volatiles emitted during their first and second callings. The ratio of (R)- to (S)-2-butanol (R/S) was 78:22 at the first calling, but shifted to 39:61 at the second calling. While investigating the composition of the female pheromones, the question arose as to whether the male preferences change in response to the shift in female pheromone composition. To answer this question, we observed the behaviour of young and old males in response to various R/S ratios as lures in the laboratory and in the field. In the flight tunnel assay of laboratory-reared individuals, young males touched female models with a 9:1 R/S ratio lure less than those with pure (R)-2-butanol; however, older males touched the two groups with equivalent frequency. In the field trap test, older males were much more attracted to (R)-2-butanol-scented lures. When we tested using lures with the same amount of (R)-2-butanol but added different amounts of the (S)-isomer, we found that increased levels of (S)-2-butanol resulted in lower attractiveness to males. (S)-2-butanol was confirmed to have an inhibitive activity in the attractiveness of (R)-2-butanol.

Age-dependent changes in the ratio of (R)- and (S)-2-butanol released by virgin females of Dasylepida ishigakiensis (Coleoptera: Scarabaeidae)

The females of the white grub beetle, Dasylepida ishigakiensis, release two enantiomers of 2-butanol, (R)-2-butanol and (S)-2-butanol. The ratio describing the relative proportions of these two enantiomers (R/S ratio) has not yet been investigated. (R)-2-Butanol has been shown to attract males in laboratory and field experiments, whereas (S)-2-butanol tends to inhibit them. To determine the R/S ratio of the 2-butanol emitted by virgin females, we collected 2-butanol from young (53 days old), mature (63 days old) and old females (73 days old) using water, extracted with an SPME fibre and subsequently injected into GC-MS. The major component of the 2-butanol emitted by the young females was (R)-2-butanol, but as the females aged, the component ratio favoured (S)-2-butanol. Young females released an 80:20 mixture of (R)- and (S)-2-butanol, whereas old females released a 45:55 mixture. The EAG response of male antennae to a 50:50 ratio (racemic mixture) showed a similar dose-response curve to that of (R)-2-butanol. The male orientation responses to (R)-2-butanol decreased when the relative proportion of (S)-2-butanol increased. An inhibitory and/or masking effect of (S)-2-butanol on male orientation behaviour was also observed in the flight tunnel assay. These results suggest that males are more strongly attracted to young females than to old females. We also discuss the possibility of using 2-butanol isomers as a control or monitoring agent for this insect.