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Apolygus lucorum (Meyer-Dür) (Hemiptera: Miridae) is a serious pest of cotton, jujube, grape and many other crops around the world. Understanding how olfactory information directs this insect to its host plants may provide environment-friendly approaches to the control of its population in agriculture. In our study, we cloned an odorant receptor gene, AlucOR46, that was specifically expressed in antennae and female-biased. Functional expression of AlucOR46 in Xenopus oocytes showed that it is tuned to six plant volatiles (S)-(-)-Limonene, (R)-(+)-Limonene, (E)-2-Hexenal, (E)-3-Hexenol, 1-Heptanol and (1R)-(-)-Myrtenol. Electroantennogram (EAG) recordings revealed that all six compounds could elicit electrophysiological responses from the antennae of A. lucorum, higher in females. Our results are in agreement with previous reports showing that (E)-2-Hexenal could attract female A. lucorum in behavior experiments. These results suggest that AlucOR46 might play an important role in locating the host plants of A. lucorum and therefore represents a suitable target for green pest control.
Figure 1. Alignment of amino acid sequences of AlucOR46, AlucOR12, AlucOR18, AlucOR30, AlucOR28 and AlucOrco. The seven transmembrane domains (TM1âTM7) are marked by solid lines. The conserved amino acid sites among the 6 ORs are marked with black shading. Amino acid similarities are very poor between these members. In particular, AlucOR46 is 12.7% identical to AlucOrco and shares 15.7%, 13.7%, 14.2% and 10.4% amino acids with AlucOR12, AlucOR18, AlucOR28 and AlucOR30, respectively.
Figure 2. Tissue expression patterns of AlucOR46 in adults of A. lucorum. A: antenna; H: heads without antenna; T: thoraxes; Ab: abdomens; L: legs. Asterisk indicates significant difference between female and male.
Figure 3. Functional characterization of AlucOR46/Orco in Xenopus oocytes. (A) Inward current responses of AlucOR46/Orco Xenopus oocytes to 10â4 M solution of (S)-(â)-Limonene, (R)-(+)-Limonene, (E)-2-Hexenal, (E)-3-Hexenol, 1-Heptanol and (1R)-(â)-Myrtenol; (B) Response profile of AlucOR46/Orco Xenopus oocytes. Error bars indicate standard error of the mean (SEM) (n = 6); (C) Tuning curve of AlucOR46. Tuning curve for the AlucOR46 to an odor panel comprising 65 odorants arranged along the x-axis. The odors which elicited the strongest responses are in the middle of the distribution, the weakest near the edges.
Figure 4. Dose-response of AlucOR46/Orco expressed in Xenopus. (A) AlucOR46/Orco Xenopus oocytes were stimulated with a concentrations range of (E)-2-Hexenal, (E)-3-Hexenol, 1-Heptanol and (1R)-(â)-Myrtenol; (B) Dose-response curves of AlucOR46/Orco Xenopus oocytes to (E)-2-Hexenal, (E)-3-Hexenol, 1-Heptanol and (1R)-(â)-Myrtenol. Responses are normalized by defining the maximal response as 100 in each group. The Error bar indicates SEM (n = 6).
Figure 5. Relative electroantennogram (EAG) responses of female and male A. lucorum to six plant volatiles. NS indicates that there are no significant differences. Asterisks indicate significant differences in EAG response between female and male antennae, p < 0.05. Error bars indicate SEM (n = 6).
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