Oviposition holds crucial significance for insect reproduction. Nevertheless, the research on the neural conduction mechanism of oviposition is still rather limited in most agricultural pests. Here, we demonstrate that the conserved Kainate receptors (KARs) expressed in the glutamatergic neurons (GNs) and the ovipositor neuromuscular junctions (NMJs) regulate the oviposition behavior in Bactrocera dorsalis . We identified two KARs (Grik2b and Grik2c), which control the oviposition behavior by influencing both oviposition preference and egg-laying quantity. Protein-ligand interaction indicated that glutamate serves as the neurotransmitter of Grik2b and Grik2c. Knockdown glutamate-coding genes adversely impacted oviposition preference and egg-laying quantity. Specific knockdown Grik2b (or Grik2c) in the GNs and NMJs could respectively influence oviposition preference and egg-laying quantity. Finally, inhibitors of KARs were screened for their ability to inhibit oviposition. Our study provides strong supporting evidence that a novel neural conduction mechanism for oviposition by uncovering the diverse roles of KARs and provides potential molecular target controlling insect oviposition.