Integrated transcriptome sequencing and RNA interference reveals molecular changes in Diaphorina citri after exposure to validamycin

Validamycin has been widely used as a specific competitive inhibitor of tre-halase.In our previous research,validamycin significantly inhibited trehalase activity and chitin synthesis in Diaphorina citri,resulting in abnormal phenotypes.However,the mechanism of validamycin's action on D.citri remains unclear.Here,using a comparative transcriptome analysis,464 differentially expressed genes(DEGs)in D.citri were identified after validamycin treatment.A Gene Ontology enrichment analysis revealed that these DEGs were mainly involved in“small molecule process”,“structural molecule ac-tivity”and“transition metal ion binding”.DEGs involved in chitin metabolism,cuticle synthesis and insecticide detoxification were validated by reverse transcription quantitative polymerase chain reaction.The RNA interference of D.citri chitinase-like protein EN03 and D.citri cuticle protein 7 genes significantly affected D.citri molting.Moreover,the recombinant chitinase-like protein EN03 exhibited a chitin-binding property,and an antimicrobial activity against Bacillus subtilis.This study provides a first insight into the molecular changes in D.citri after exposure to validamycin and identifies two effective RNA interference targets for D.citri control.