A novel method for molecular targeting of insecticide resistance in Rhopalosiphum padi L. (Homoptera: Aphididae)

The bird cherry-oat aphid (Rhopalosiphum padi L.) is a devastating cereal pest that develops high resistance to organophosphate and carbamate insecticides. Because acetylcholinesterase (ACE) is the target of carbamate and organophosphate insecticides, the resistance mechanism usually involves mutations occurring in ACE-encoding genes, Ace1 and Ace2. Here, we describe a novel polymerase chain reaction (PCR)-based method for the diagnosis of resistance cases associated with the point mutation F368L in the Ace2 gene. We amplified a 127bp DNA fragment from Ace2 gene using a modified reverse primer, and digested the amplification product using SmaI endonuclease. This procedure enabled a simple and rapid distinction between resistant and susceptible genotypes for F368L mutation. Subsequently, we screened 152 R. padi samples, and found that F368L mutation occurred at low frequency, in both the homozygous (R/R) and heterozygous (R/S) states. Based upon the results of this study, we believe that molecular diagnosis of insecticide resistance should be generalized to genes and mutations involved in this process, toward an optimal accuracy of insecticide applications.