Functional Identification of Px-fringe and Px-engrailed Genes under Heat Stress in Chlorpyrifos-Resistant and -Susceptible Plutela xylostella (Lepidoptera: Plutellidae)

Abstract

In our previous research, the fitness cost of resistance of the diamondback moth (DBM), Plutella xylostella found in insecticide-resistant DBM (Rc-DBM) under heat stress was based on heavier damage to wing veins when compared to insecticide-susceptible DBM (Sm-DBM). To investigate the molecular mechanism of the damage to the veins between Rc- and Sm-DBM, the full-length sequences of two related genes involved in the development of wing veins, fringe (Px-fng) and engrailed (Px-en) of DBM were cloned, and the mRNA expressions of both Px-fng and Px-en were studied. The Px-fng and Px-en cDNA contained 1038 bp and 1152 bp of open reading frames (ORFs), respectively, which encoded a putative protein comprising 345 and 383 amino acids with a calculated molecular weight of 39.59 kDa and 42.69 kDa. Significantly down regulated expressions of Px-fng and Px-en under heat stress were found in pupae and adults of Rc-DBM compared to Sm-DBM, and a result of higher damage to wing veins in Rc-DBM under heat stress. Based on RNAi experiments, significant inhibitions on expressions of Px-fng and Px-en in both Sm-DBM and Rc-DBM were found when the pupae were infected by dsFng or dsEn. Corresponding to these, infections of dsFng or dsEn resulted in significant decrease of eclosion rate and increase malformation rate of DBM. Our results suggest that the higher damage of wing veins in DBM might be related to the heavier inhibitions of Px-fng and Px-en expression, and the Px-fng and Px-en are involved in the development of wings and veins.