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Ratios of compounds in host plant odors fluctuate with the phenological stage of the plant. In the present study, we investigated the effect of changing ratios of host plant volatile constituents on herbivore insect attraction and olfactory information processing. We tested a synthetic mixture of bioactive peach shoot volatiles with different concentrations of one of the mixture constituents, benzonitrile, on oriental fruit moth Cydia (=Grapholita) molesta females. Y-tube olfactometer bioassays showed that female attraction to the mixture was maintained while increasing the benzonitrile level up to 100 times. Further increases led to behaviorally ineffective mixtures. Then, we recorded odor-evoked neural activity patterns in the antennal lobes, the main olfactory center of the brain, using calcium imaging. Benzonitrile-containing mixtures elicited strong activation in two glomeruli, which were found to process mixture-related information in specific ways. Activation in one glomerulus directly paralleled behavioral effects of the different ratios tested whereas a deviating pattern was noted in the other glomerulus. Our results indicate that the ratio of constituents in a volatile mixture can be varied to a certain degree without reducing female attraction. Thus, volatile blends in nature might vary quantitatively within a certain range without affecting odor-guided host location. Neurophysiological results showed that the processing of mixture-related information inside the antennal lobes is not uniform across glomeruli. Thus, final processing of this information probably takes place in higher-order brain centers.

Behavioral and neurophysiological responses of an insect to changing ratios of constituents in host plant-derived volatile mixtures