Quantitative and qualitative variation in the mRNA for carboxylesterases in insecticide-susceptible and resistant Myzus persicae (Sulz)

Abstract

RNA was extracted from two insecticide-resistant clones and one susceptible clone of M. persicae. The resistant clones each produced large amounts of one of two closely related carboxylesterases, the enzymes responsible for cross-resistance to a wide range of insecticides. After purification by affinity chromatography on oligo(dT) cellulose, the mRNA was translated into protein in a rabbit reticulocyte lysate system with [ l- 35S]methionine. The resultant radiolabelled esterases were immunoprecipitated from the products with IgG prepared from an antiserum to one form of the enzyme, but cross-reacting with both. The bound enzyme was extracted by affinity chromatography on protein A sepharose, and characterized alongside the total radiolabelled proteins by SDS electrophoresis and fluorography. The translation products of the two resistant clones each contained large amounts of an immunoprecipitable protein. However, no such protein was detected in the translation products of the mRNA from susceptible aphids showing that resistant aphids produce much more of the mRNA encoding the enzymes responsible for resistance. It was also shown that the enzymes from the two resistant clones had primary structures differing from each other by 1 kDa. In addition, the nascent forms of both enzymes differed from their native forms by 8 kDa and glycosylation was shown to be responsible for this post-translational modification. The likely genetic basis of the changes in mRNA is discussed and related to the karyotype of the resistant clones.