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RNA interference (RNAi) is an effective gene-silencing tool, and double stranded RNA (dsRNA) is considered a powerful strategy for gene function studies in insects. In the present study, we aimed to investigate the function of trehalase (TRE) genes ( TRE 1-1 ,  TRE 1-2 , and  TRE-2 ) isolated from the brown planthopper  Nilaparvata lugens , a typical piercing-sucking insect in rice, and investigate their regulating roles in chitin synthesis by injecting larvae with dsRNA. The results showed that  TRE1  and  TRE2  had compensatory function, and the expression of each increased when the other was silenced. The total rate of insects with phenotypic deformities ranged from 19.83 to 24.36% after dsTRE injection, whereas the mortality rate ranged from 14.16 to 31.78%. The mRNA levels of genes involved in the chitin metabolism pathway in RNA-Seq and DGEP, namely hexokinase ( HK ), glucose-6-phosphate isomerase ( G6PI ) and chitinase ( Cht ), decreased significantly at 72 h after single dsTREs injection, whereas two transcripts of chitin synthase ( CHS ) genes decreased at 72 h after dsTRE1-1 and dsTREs injection. These results demonstrated that  TRE  silencing could affect the regulation of chitin biosynthesis and degradation, causing moulting deformities. Therefore, expression inhibitors of  TRE s might be effective tools for the control of planthoppers in rice.

Functional characterization of three trehalase genes regulating the chitin metabolism pathway in rice brown planthopper using RNA interference