Odorant molecules interact with odorant receptors (ORs) lining the pores on the surface of the sensilla on an insect’s antennae and maxillary palps. This interaction triggers an electrical signal that is transmitted to the insect’s nervous system, thereby influencing its behavior. Orco, an OR coreceptor, is crucial for olfactory transduction, as it possesses a conserved sequence across the insect lineage. In this study, we focused on 2,4-di-tert-butylphenol (DTBP), a single substance present in acetic acid bacteria culture media. We applied DTBP to oocytes expressing various Drosophila melanogaster odor receptors and performed electrophysiology experiments. After confirming the activation of DTBP on the receptor, the binding site was confirmed through point mutations. Our findings confirmed that DTBP interacts with the insect Orco subunit. The 2-heptanone, octanol, and 2-hexanol were not activated for the Orco homomeric channel, but DTBP was activated, and the EC₅₀ value was 13.4 ± 3.0 μM. Point mutations were performed and among them, when the W146 residue changed to alanine, the E_max value was changed from 1.0 ± 0 in the wild type to 0.0 ± 0 in the mutant type, and all activity was decreased. Specifically, DTBP interacted with the W146 residue of the Orco subunit, and the activation manner was concentration-dependent and voltage-independent. This molecular-level analysis provides the basis for novel strategies to minimize pest damage. DTBP, with its specific binding to the Orco subunit, shows promise as a potential pest controller that can exclusively target insects.

The predatory gall midge, Feltiella acarisuga (Vallot) (Diptera: Cecidomyiidae), is an acarivorous species that mainly feeds on spider mites (Acarina: Tetranychidae). Because of its cosmopolitan distribution and predation efficacy, it is considered an important natural enemy available as a biological agent for augmentative biocontrol. However, despite its practical use, the thermal development and survival response to temperature have not yet been fully studied. In this study, we investigated the stage-specific development and survival of F. acarisuga at seven temperatures (11.5, 15.7, 19.8, 23.4, 27.7, 31.9, and 35.4 °C) to examine the effect of temperature on its lifecycle. All developmental stages could develop at 11.5–31.9 °C, but the performance was different according to the temperature. From the linear development rate models, the lower development threshold and thermal constant of the total immature stage were estimated at 8.2 °C and 200 DD, respectively. The potential optimal and upper threshold temperatures for the total immature stage were estimated as 29.3 and 35.1 °C using a non-linear development model. The operative thermal ranges for development and survival at 80% of the maximum rate were 24.5–32.3 and 14.7–28.7 °C, respectively. Thus, it was suggested that 24.5–28.7 °C was suitable for the total immature stage. In contrast, conditions around 8 °C and 35 °C should be avoided due to the lower development rate and high mortality. Our findings provide fundamental information for an effective mass-rearing and releasing program of F. acarisuga in an augmentative biocontrol program and help to predict phenology.

It is known that two-spotted spider mite (Tetranychus urticae), a major pest of strawberry, is difficult to manage. This study was conducted to determine the optimal sampling unit to increase management efficiency of T. urticae in strawberries. The sampling unit was determined by characterizing within-strawberry distribution of T. urticae and by comparing coefficient of variation (CV) and correlation coefficient (r²) among potential sampling units. There was a significant (p < 0.05) difference in densities within a strawberry according to the leaf age. However, there was no significant difference in its density within a trifoliate leaf. More T. urticae were found on young-fully-opened (moderately old) leaves than on young and old ones. Moreover, these leaves had lower CV and r² values than others. More specifically, optimal sampling units of T. urticae were fifth, fifth or sixth, and sixth oldest leaves for motiles (immatures and adults), all stages (motiles and eggs), and eggs, respectively. The required sampling number should be determined depending on the density. However, for management purposes, one and two leaflets would be acceptable for eggs and motiles, respectively. By using this small number of suitable sampling unit, the sampling and management for T. urticae in strawberries could be more efficient than before.

The insect olfactory system is vital for survival, enabling the recognition and discrimination of a wide range of odorants present in the environment. This process is mediated by odorant receptors (Ors) and the highly conserved co-receptor Orco. Insect Ors are structurally distinct from mammalian olfactory receptors, a divergence that presents unique advantages for developing insect-specific pest control strategies. In this study, we explored the molecular-level interactions between insect Ors and volatile organic compounds. Specifically, we investigated the response of Ors/Orco to phenethyl acetate (PA), a volatile compound found in the culture media of acetic acid bacteria. PA elicited activation in a concentration-dependent, reversible, and voltage-independent manner in Or1a, Or24a, and Or35a when combined with Orco, as well as in Orco homomers. Through molecular docking studies, we determined that the PA-binding site is localized to the Orco subunit, a highly conserved protein across diverse insect taxa. To further elucidate the role of key residues in the Orco homotetramer receptor, we performed site-directed mutagenesis. A mutational analysis identified W146 and E153 as critical residues for PA binding and activation. A double-mutant Orco receptor (W146A + E153A) exhibited a significant reduction in PA-induced activation compared to the wild-type receptor. These findings indicate that PA functions as an agonist for the Drosophila melanogaster Orco receptor and highlight its potential applications in chemosensory research and insect pest management strategies.