To better understand the olfactory mechanisms in the rice leaf folder, Cnaphalocrocis medinalis (Guenée), a serious pest of rice in Asia, we established six partial transcriptomes from antennae, protarsus, and reproductive organs of male and female adults. A total of 102 transcripts were identified, including 29 odorant receptors (ORs), 15 ionotropic receptors (IRs), 30 odorant-binding proteins (OBPs), 26 chemosensory proteins (CSPs), and 2 sensory neuron membrane proteins (SNMPs). The expression patterns of these genes were calculated by fragments per kilobase of exon per million fragments mapped (FPKM) and validated by real-time quantitative PCR (RT-qPCR). Some transcripts were exclusively expressed in specific organs, such as female protarsus, whereas others were universally expressed, this varied expression profile may provide insights into the specific functions mediated by chemoreception proteins in insects. To the best of our knowledge, among the 102 identified transcripts, 81 are novel and have never been reported before. In addition, it also is the first time that ORs and IRs are identified in C. medinalis. Our findings significantly enhance the currently limited understanding olfactory mechanisms of the olfactory mechanisms underlying the chemoreception system in C. medinalis.

It is well known that the odorant-binding proteins (OBPs) play crucial roles in insect olfactory detection. To explore if interactions of OBPs affect olfactory coding in the rice leaffolder Cnaphalocrocis medinalis ligand-binding experiments, molecular docking, RNA interference and electrophysiological recording were performed. The binding activity of two C. medinalis OBPs (CmedOBPs) to rice plant volatiles showed wide flexibility depending on the structure of ligands and interactions of CmedOBPs involved. The binding sites of CmedOBP2 and CmedOBP3 to rice plant volatiles were well predicted by three-dimensional structure modelling and molecular docking experiments. In addition, the interactions of these two CmedOBPs in the perception of rice volatiles were demonstrated by RNA interference experiments. When a single double-stranded RNA (dsRNA)-CmedOBP2 was injected, the expression of CmedOBP2 was significantly reduced and the expression of CmedOBP3 was significantly increased, and vice versa. When both dsRNA-CmedOBP2 and 3 were injected together, greater reduction of electroantennogram responses to rice plant volatiles was induced than that seen with individual injection of either dsRNA-CmedOBP2 or dsRNA-CmedOBP3. These results clearly indicate that the interactions of CmedOBP2 and CmedOBP3 have significant effects on C. medinalis during the detection of host plant volatiles.

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