Bactrocera dorsalis is a notorious pest widely distributed across most Asian countries. With the rapid development of pesticide resistance, new pest control methods are urgently needed. RNAi-based sterile insect technique (SIT) is a species-specific pest management strategy for B. dorsalis control. It is of great significance to identify more target genes from B. dorsalis, and improve the RNAi efficiency. In this study, microinjection-based RNAi results showed that six 20E response genes were necessary for male fertility of B. dorsalis, of which E75 was identified as the key target according to the lowest egg-laying number and hatching rate after E75 knockdown. Three nanoparticles chitosan (CS), chitosan‑sodium tripolyphosphate (CS-TPP), and star polycation (SPc) were used to encapsulate dsE75 expressed by HT115 strain. Properties analysis of nanoparticle-dsRNA complexes showed that both CS-TPP-dsRNA and SPc-dsRNA exhibited better properties (smaller size and polydispersity index) than CS-dsRNA. Moreover, oral administration of CS-TPP-dsE75 and SPc-dsE75 by males resulted in more abnormal testis and significantly lower fertility than feeding naked dsE75. Semi-field trials further confirmed that the number of hatched larvae was dramatically reduced in these two groups. Our study not only provides more valuable targets for RNAi-based SIT, but also promotes the application of environment-friendly management against B. dorsalis in the field.Copyright © 2024. Published by Elsevier B.V.