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. 2021 Jan 20:11:624287.
doi: 10.3389/fphys.2020.624287. eCollection 2020.

Function Analysis of P450 and GST Genes to Imidacloprid in Aphis craccivora (Koch)

Yuan-Xue Yang  1 , Rong-Hua Lin  2 , Zhuo Li  1 , Ai-Yu Wang  1 , Chao Xue  1 , Ai-Ling Duan  1 , Ming Zhao  1 , Jian-Hua Zhang  3
Affiliations

Function Analysis of P450 and GST Genes to Imidacloprid in Aphis craccivora (Koch)

Yuan-Xue Yang et al. Front Physiol. .

Abstract

Aphis craccivora (Koch) is an economically important pest that affects legumes in worldwide. Chemical control is still the primary efficient method for A. craccivora management. However, the mechanism underlying insecticide resistance in A. craccivora has not been elucidated. A previous study observed that piperonyl butoxide (PBO) and diethyl maleate (DEM) significantly synergized imidacloprid in A. craccivora field populations, indicating that cytochrome P450 (P450) and glutathione S-transferase (GST) genes may play pivotal roles in imidacloprid resistance. In this study, 38 P450 genes and 10 GST genes were identified in A. craccivora through transcriptomic analysis. The expression levels of these P450 and GST genes were measured in susceptible (SUS) strains of A. craccivora under imidacloprid treatment with LC15, LC50, and LC85 doses. The expression levels of CYP18A1, CYP6CY21, CYP6DA1, CYP6DA2, CYP4CJ1, CYP4CJ2, and CYP380C6 were up-regulated in the three treatments. Most of these genes belong to CYP3 and CYP4 Clans. In addition, the expression levels of all P450 and GST genes in A. craccivora were also measured in the Juye (JY) and Linqing (LQ) field populations. The expression levels of CYP6DA2, CYP4CJ1, and CYP380C6 were up-regulated in the SUS strain after imidacloprid treatment at three doses, and these genes were overexpressed in the JY population. Furthermore, the sensitivity of A. craccivora to imidacloprid was significantly increased after knockdown of CYP380C6 and CYP6DA2 through RNA interference. These results may help to elucidate the mechanisms underlying of imidacloprid resistance in A. craccivora.

Keywords: Aphis craccivora; GST genes; P450 genes; RNA interference; expression induction; imidacloprid.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Phylogenetic analysis of P450 genes. The numbers above the branches indicate the support for the phylogenies, and only values above 50% are shown. Blackened circles indicate the Aphis craccivora position. Ac, A. craccivora; Ag, Aphis gossypii; Ap, Acyrthosiphon pisum; Nl, Nilaparvata lugens.
Figure 2
Figure 2
Phylogenetic analysis of GST genes. The numbers above the branches indicate the support for the phylogenies, and only values above 50% are shown. Blackened circles indicate the A. craccivora position. Ac, A. craccivora; Ag, Aphis gossypii; Ap, Acyrthosiphon pisum; Nl, Nilaparvata lugens.
Figure 3
Figure 3
Fold changes in the expression of 38 P450 genes and 10 GST genes in the A. craccivora susceptible strain after the LC15 dose of imidacloprid treatment. The significant differences were marked by asterisks. *Significantly at 0.05 level. The ns represents no significant difference. (A) Fold changes in the expression of P450 genes from CYP2 and Mito Clan. (B) Fold changes in the expression of P450 genes from CYP4 Clan. (C) Fold changes in the expression of P450 genes from CYP3 Clan. (D) Fold changes in the expression of GST genes.
Figure 4
Figure 4
Fold changes in the expression of 38 P450 genes and 10 GST genes in the A. craccivora susceptible strain after the LC50 dose of imidacloprid treatment. The significant differences were marked by asterisks. *Significantly at 0.05 level. The ns represents no significant difference. (A) Fold changes in the expression of P450 genes from CYP2 and Mito Clan. (B) Fold changes in the expression of P450 genes from CYP4 Clan. (C) Fold changes in the expression of P450 genes from CYP3 Clan. (D) Fold changes in the expression of GST genes.
Figure 5
Figure 5
Fold changes in the expression of 38 P450 genes and 10 GST genes in the A. craccivora susceptible strain after the LC85 dose of imidacloprid treatment. The significant differences were marked by asterisks. *Significantly at 0.05 level. The ns represents no significant difference. (A) Fold changes in the expression of P450 genes from CYP2 and Mito Clan. (B) Fold changes in the expression of P450 genes from CYP4 Clan. (C) Fold changes in the expression of P450 genes from CYP3 Clan. (D) Fold changes in the expression of GST genes.
Figure 6
Figure 6
Fold changes of LQ and JY vs. SUS in the expression levels of 10 GST genes the in A. craccivora. The significant differences were marked by asterisks. *Significantly at 0.05 level. The ns represents no significant difference. (A) Fold changes in the expression of P450 genes from CYP2 and Mito Clan. (B) Fold changes in the expression of P450 genes from CYP4 Clan. (C) Fold changes in the expression of P450 genes from CYP3 Clan. (D) Fold changes in the expression of GST genes.
Figure 7
Figure 7
The dsRNA-mediated suppression of P450 genes and effects on imidacloprid toxicity in A. craccivora SUS strain. (A) The dsRNA-mediated suppression the expression of three P450 genes in A. craccivora SUS strain fed on the artificial diet with dsRNA. (B) The mortality of A. craccivora strain with LC50 dose of imidacloprid treatment after knockdown of CYP6DA2, CYP4CJ1, and CYP380C6. The significant differences were marked by asterisks. *Significantly at 0.05 level. The ns represents no significant difference. IMI, imidacloprid.
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