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. 2020 May 1;20(3):12.
doi: 10.1093/jisesa/ieaa047.

Silencing of Cytochrome P450 in Spodoptera frugiperda (Lepidoptera: Noctuidae) by RNA Interference Enhances Susceptibility to Chlorantraniliprole

Zhang Bai-Zhong  1 , Su Xu  1 , Zhen Cong-Ai  2 , Lu Liu-Yang  1 , Li Ya-She  1 , Ge Xing  1 , Chen Dong-Mei  1 , Pei Zhang  1 , Shi MIng-Wang  1 , Chen Xi-Ling  1
Affiliations

Silencing of Cytochrome P450 in Spodoptera frugiperda (Lepidoptera: Noctuidae) by RNA Interference Enhances Susceptibility to Chlorantraniliprole

Zhang Bai-Zhong et al. J Insect Sci. .

Abstract

Fall armyworm, Spodoptera frugiperda (Smith), has caused significant losses for crop production in China. The fall armyworm is mainly controlled by the chemical insecticides, whereas the frequent application of insecticides would result in the resistance development. Insect cytochrome P450 monooxygenases play an essential part in the detoxification of insecticides. In this study, five P450 genes were selected to determine the role in response to insecticides by RNA interference (RNAi). Developmental expression pattern analysis revealed that S. frugiperda CYP321A8, CYP321A9, and CYP321B1 were highest in second-instar larvae among developmental stages, with 2.04-, 3.39-, and 8.58-fold compared with eggs, whereas CYP337B5 and CYP6AE44 were highest in adult stage, with 16.3- and 10.6-fold in comparison of eggs, respectively. Tissue-specific expression pattern analysis exhibited that CYP321A8, CYP321B1, and CYP6AE44 were highest in the midguts, with 3.56-, 3.33-, and 3.04-fold compared with heads, whereas CYP321A9 and CYP337B5 were highest in wings, with 3.07- and 3.36-fold compared with heads, respectively. RNAi was also conducted to explore detoxification effects of the five P450 genes on chlorantraniliprole. The second-instar larvae became more sensitive to chlorantraniliprole with a higher mortality rate than the control, after silencing CYP321A8, CYP321A9, and CYP321B1, respectively. These findings strongly supported our viewpoint that CYP321A8, CYP321A9, and CYP321B1 may play a critical role in insecticide detoxification. It will provide a basis for further study on regulation of P450 genes and the management of S. frugiperda.

Keywords: Spodoptera frugiperda; RNAi; cytochrome (P450); insecticide detoxification.

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Figures

Fig. 1.
Fig. 1.
Developmental expression levels of the five P450 genes in Spodoptera frugiperda. Data are means ± SE of three biological replicates. The relative expression was calculated using the 2−ΔΔCT method based on the value of the egg expression, which was ascribed an arbitrary value of 1. The bars with lowercase letters are significantly different according to the one-way ANOVA, followed by Tukey’s multiple comparison test (P < 0.05).
Fig. 2.
Fig. 2.
The tissue expression profiles of the five P450 genes in Spodoptera frugiperda. The relative expression was calculated using the 2−ΔΔCT method based on the expression value of the adults head, which was assigned an arbitrary value of 1. The bars with lowercase letters are significantly different according to the one-way ANOVA, followed by Tukey’s multiple comparison test (P < 0.05).
Fig. 3.
Fig. 3.
Relative expression levels of the target P450 gene and the mortality (%) of Spodoptera frugiperda at the 1 mg/l chlorantraniliprole. (A) Relative expression of the target P450 gene and (B) mortality (%) of Spodoptera frugiperda at 1 mg/l chlorantraniliprole. The data were expressed as the means ± SE. The bars with lowercase letters are significantly different according to the one-way ANOVA, followed by Tukey’s multiple comparison test (P < 0.05).
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