PROSPECÇÕES DE MUTAÇÕES NO GENE CITOCROMO P450 CYP12A2 EM MOSCA-DOS-ESTÁBULOS RELACIONADAS A RESISTÊNCIA AOS INSETICIDAS PIRETROIDES

Abstract

The stable fly is a hematophagous parasite that causes great damage to the sugarcane and livestock production chains. It is known that pesticide resistance in insects is an evolutionary phenomenon and there are different types of genetic mechanisms that can cause a reduction in sensitivity to insecticides, due to changes in the genetic code (mutations) that can affect the target site of insecticides and/or their metabolism. The study aims to prospect mutations in the cytochrome P450 CYP12A2 gene that may be related to insecticide resistance in order to contribute to the effective management and control of this pest. A total of 157 flies were genotyped and distributed in two independent experiments. The primers used in both experiments were designed from the reference sequence NW_013173580. In the first study, the Sanger methodology was used to evaluate the existence of mutations in the region delimited by the first three exons of the gene. The DNA of resistant (16) and susceptible (20) flies obtained by bioassay, as well as the DNA of control samples from the permanent colony of CNPGC (6) and susceptible samples from the colony of Kerville – USA (3) were extracted, amplified, sequenced and genotyped individually. Three mutations were observed in the evaluated region, being a SNP in the second exon (g.330343T>G), an insertion of a thymine in a mononucleotide composed of eight thymines (g.330531-330532insT) in the first intron and a SNP in the 1st exon (g.330702G>C). There was no significant difference between the genotyped populations (p