Pheromones of three ambrosia beetles in the Euwallacea fornicatus species complex: ratios and preferences

Abstract

Three cryptic species in the Euwallacea fornicatus species complex were reared in laboratory colonies and investigated for the presence of pheromones. Collections of volatiles from combinations of diet, fungus, beetles, and galleries from polyphagous shot hole borer (Euwallacea sp. #1) revealed the presence of 2-heneicosanone and 2-tricosanone only in the presence of beetles, regardless of sex. Subsequent examination of volatiles from the other two species, tea shot hole borer (Euwallacea sp. #2) and Kuroshio shot hole borer (Euwallacea sp. #5), revealed these two ketones were present in all three species but in different ratios. In dual choice olfactometer behavioral bioassays, mature mated females were strongly attracted to the binary blend of ketones matching their own natural ratios. However, females in each species were repelled by the ketone blends in ratios corresponding to the other two species. Males of each species responded similarly to females when presented with ratios matching their own or the other two species. The presence of these compounds in the three beetle species, in ratios unique to each species, and their strong species-specific attraction and repellency, suggests they are pheromones.The ecological function of these pheromones is discussed. In addition to the pheromones, the previously known attractant (1S,4R)-p-menth-2-en-1-ol (also known as quercivorol) was discovered in the presence of the fungal symbionts, but not in association with the beetles.Quercivorol was tested in a dual-choice olfactometer and was strongly attractive to all three species. This evidence suggests quercivorol functions as a kairomone for members of the E. fornicatus species complex, likely produced by the symbiotic fungi. 29 #5), revealed these two ketones were present in all three species but in different ratios. In dual 30 choice olfactometer behavioral bioassays, mature mated females were strongly attracted to a 31 synthetic binary blend of ketones matching their own natural ratios. However, females in each 32 species were repelled by ketone blends in ratios corresponding to the other two species. Males 33 of each species responded similarly to females when presented with ratios matching their own or 34 the other two species. The presence of these compounds in the three beetle species, in ratios 35 unique to each species, and their strong species-specific attraction and repellency, suggests they 36 are pheromones. The ecological function of these pheromones is discussed. In addition to the 37 pheromones, the previously known attractant (1S,4R)-p-menth-2-en-1-ol (also known as 38 quercivorol) was discovered in the presence of the fungal symbionts, but not in association with 39 the beetles. Quercivorol was tested in a dual-choice olfactometer and was strongly attractive to 40 all three species. This evidence suggests quercivorol functions as a kairomone for members of 41 the E. fornicatus species complex, likely produced by the symbiotic fungi.