Acta Entomologica Sinica ›› 2020, Vol. 63 ›› Issue (7): 843-850.doi: 10.16380/j.kcxb.2020.07.008

• RESEARCH PAPERS • Previous Articles     Next Articles

Construction of transgenic Drosophila strains with carboxylesterase genes of Locusta migratoria (Orthoptera: Acrididae) and their roles in the metabolic detoxification of organophosphorus pesticides

YIN Fei1, YANG Yang2, ZHANG Xu-Bo2, ZHANG Jian-Zhen2, ZHANG Jian-Qin1,*   

  1.  (1. Modern Research Center for Traditional Chinese Medicine, Shanxi University, Taiyuan 030006, China; 2. Institute of Applied Biology, Shanxi University, Taiyuan 030006, China)
  • Online:2020-07-20 Published:2020-07-29

Abstract:  【Aim】 To determine whether the carboxylesterase genes LmCesA1 and LmCesA2 of Locusta migratoria are involved in the metabolic detoxification of organophosphorus insecticides in vivo. 【Methods】 Two transgenic strains of Drosophila melanogaster were constructed by using Gal4/UAS system and transgenic technology. Three Gal4 strains (act-Gal4, tub-Gal4 and c601-Gal4) were selected to hybridize with transgenic Drosophila strains (UAS-LmCesA1 and UAS-LmCesA2) and the reference strain (RB0006{y v; attP40, y+}). The offspring of transgenic strains was verified at the DNA and RNA levels, and then the successfully constructed strains were screened out. The resistance of the offspring of the transgenic strains and Gal4 strains to malathion was detected by bioassay. 【Results】 The identification results of transgenic Drosophila at the DNA level showed that the target genes LmCesA1 and LmCesA2 were amplified in the transgenic strains tub>LmCesA1 and tub>LmCesA2, respectively, but not in the reference group (tub>attP40), while those at the RNA level revealed that the two genes were expressed in the corresponding hybrid offspring, indicating that the transgenic Drosophila strains were successfully constructed. Tissue expression profiles of the two target genes in adults of the transgenic Drosophila showed that LmCesA1 and LmCesA2 were highly expressed in the gut of the transgenic strains c601>LmCesA1 and c601>LmCesA2, respectively. In the transgenic strain c601>LmCesA1 the expression level of LmCesA1 in the gut was 7.6- and 16.7-fold as high as those in the brain and cuticle, respectively, while in the transgenic strain c601>LmCesA2 the expression level of LmCesA2 in the gut was 5.4- and 10.9-fold as high as those in the brain and cuticle, respectively. The results of insecticide bioassay demonstrated that compared with the F1 generation of hybrid strain in the reference group (c601>attP40), the hybrid offspring overexpressing LmCesA2 (c601>LmCesA2) showed significantly enhanced resistance to malathion, with the resistance ratio of 1.67. 【Conclusion】 The conclusion of this study is consistent with that of our previous research by RNAi combined with insecticide bioassay, i.e., LmCesA2 may be involved in the metabolic detoxification of malathion in L. migratoria.

Key words: Locusta migratoriaDrosophila melanogaster, carboxylesterase gene, transgene, insecticide, metabolic detoxification, pesticide resistance