Electrophysiological and behavioral responses of Ips duplicatus aggregation pheromone in Inner Mongolia, China: amitinol as a potential pheromone component

Volatiles from Ips duplicatus male hindgut extracts and aeration samples of spruce logs colonized by the virgin males were analyzed by coupled gas chromatography-mass spectrometry (GC-MS), coupled gas chromatography-electroantennographic detection (GC-EAD), and field trapping bioassays in Inner Mongolia, China. GC-MS analyses showed that males from nuptial chambers (for reproduction) produced hindgut volatile profiles similar to those of maturation feeding males from the nonreproductive galleries. The known aggregation pheromone components, ipsdienol (Id) and E-myrcenol (EM) are the major constituents of this blend, followed by several minor bark beetle-related compounds: 2-methyl-3-buten-2-ol, ipsenol, cis-verbenol, verbenone, 2-phenylethanol, myrtenol, and trans-myrtanol. The enantiomeric composition of the Id from the male hindgut extracts was determined to be a racemic mixture (50/50) by enantioselective GC. GC-EAD experiments indicated that I. duplicatus antennae from both sexes responded strongly not only to the known aggregation pheromone components, Id and EM, but also to the other minor conspecific-produced compounds. The strongest antennal response was elicited by the major pheromone component, Id, which was ca. 2-3 times higher than that elicited by the second component, EM. Electroantennogram dose-responses indicated that antennal response threshold to Id was approximately 100 times lower than that of EM. No synergistic effects between these occurred at peripheral level. A strong and repeatable EAD response was found to a trace compound (flame ionization detection undetectable) from both hindgut and aeration samples. The compound was identified as amitinol (At) based on the retention time and a further EAD analysis of a synthetic mixture. In the field trapping experiments, At increased the attraction of I. duplicatus to the traps baited with a binary blend of EM/Id (1:1) in a synergistic manner when tested at a low release rate (1:1:0.1) but had no effect on catches at the high release rate (1:1:1). This result suggests that At might be a part of the Chinese I. duplicatus aggregation pheromone system. The production ratios of EM/Id from the Inner Mongolian population (EM/Id[approximately equal]1:2.0-4) were twice as high as those reported from Europe (EM/Id[approximately equal]1:9), corresponding well with the differences in the optimal response ratios