Functional characterization of two spliced variants of fructose gustatory receptor in the diamondback moth, Plutella xylostella

Abstract

Insect gustatory system plays important roles in multiple behaviors including feeding, mating, and oviposition. Gustatory receptors (GRs), located on the dendritic membrane of gustatory sensory neurons (GSNs), are crucial in peripheral coding of non-volatile compounds. However, GRs and their detailed functions remain poorly understood in lepidopteran pests. In the present work, focusing on GR genes of Plutella xylostella, an important worldwide crop pest, we cloned a candidate fructose GR gene that has two spliced variants (PxylGR43a-1 and PxylGR43a-2), and determined the tissue expression profiles by semi-quantitative reverse transcription PCR (RT-PCR). It revealed that both GR variants were highly expressed in antennae and less highly in heads of adults, while PxylGR43a-2 was also weakly expressed in other tested tissues. Functional analyses were further conducted using the Xenopus oocyte system. PxylGR43a-1 and PxylGR43a-2 both responded specifically to the d-fructose among the 12 tested sugar compounds, but PxylGR43a-2 showed much higher current response than PxylGR43a-1. In addition, the proboscis extension reflex (PER) assay was conducted, demonstrating that female moths could respond to d-fructose following stimulation of antenna. Taken together, our study contributes to elucidation of the molecular mechanisms of fructose reception and provides a potential target for development of GR based pest control techniques.