Regulation of GSTu1-mediated insecticide resistance in Plutella xylostella by miRNA and lncRNA

Author summary The development of insecticide resistance in insect pests is a worldwide concern and a major problem in agriculture. Understanding the genetics of insecticide resistance is critical for effective crop protection. Plutella. xylostella (L.), a major pest of cruciferous crops, has developed resistance to almost all kinds of insecticide, and has become one of the most resistant pests in the world. Overexpression of detoxification enzymes is closely associated with insecticide resistance, but researches on their regulatory mechanism are still very limited. Here, GSTu1 was identified to be upregulated in several chlorantraniliprole-resistant P. xylostella populations and was confirmed to be involved in chlorantraniliprole resistance by direct degradation of this insecticide. Further, lnc-GSTu1-AS transcribed from the opposite DNA strand to GSTu1 was identified to be able to enhance the mRNA stability of GSTu1 by blocking miRNA activity, and thus increased the resistance of P. xylostella to chlorantraniliprole. The results provide further insights into the mechanisms underlying metabolic resistance. The evolution of resistance to insecticides is well known to be closely associated with the overexpression of detoxifying enzymes. Although the role of glutathione S-transferase (GST) genes in insecticide resistance has been widely reported, the underlying regulatory mechanisms are poorly understood. Here, one GST gene (GSTu1) and its antisense transcript (lnc-GSTu1-AS) were identified and cloned, and both of them were upregulated in several chlorantraniliprole-resistant Plutella xylostella populations. GSTu1 was confirmed to be involved in chlorantraniliprole resistance by direct degradation of this insecticide. Furthermore, we demonstrated that lnc-GSTu1-AS interacted with GSTu1 by forming an RNA duplex, which masked the binding site of miR-8525-5p at the GSTu1-3 ' UTR. In summary, we revealed that lnc-GSTu1-AS maintained the mRNA stability of GSTu1 by preventing its degradation that could have been induced by miR-8525-5p and thus increased the resistance of P. xylostella to chlorantraniliprole. Our findings reveal a new noncoding RNA-mediated pathway that regulates the expression of detoxifying enzymes in insecticide-resistant insects and offer opportunities for the further understanding of the mechanisms of insecticide and drug resistance.