The green rice leafhopper, Nephotettix cincticeps (Hemiptera: Cicadellidae), salivary protein NcSP75 is a key effector for successful phloem ingestion

Nephotettix cincticeps, a prevalent rice pest, injects gelling and watery saliva into plant tissues during the sucking process. Certain components within the saliva are believed to interact with plant cellular constituents and play important roles in overcoming host plant defense responses. Based on our previous analysis of the salivary gland transcriptome and secreted saliva proteome of N. cincticeps, in this study, we analyzed the biological functions of salivary protein, NcSP75 (N. cincticeps salivary protein 75 kD). NcSP75, a salivary glands-specific gene, showed low similarities to any previously reported sequences. Knockdown of NcSP75 by RNA interference (RNAi) reduced the longevity of treated nymphs to approximately half of the longevity of controls and caused severe developmental retardation. Furthermore, the knockdown of NcSP75 decreased the survival rate of adults, and reduced the number of deposited eggs and hatched nymphs. Thus, the adverse effects caused by the knockdown of NcSP75 were observed throughout the lifetime of N. cincticeps, when feeding on rice plants. In contrast, no reduction was observed in the survival rate of the knockdown of NcSP75 adults fed on an artificial diet. Electrical penetration graph measurements taken from adult females feeding on rice plants showed a significantly shorter duration of phloem ingestion associated with the knockdown of NcSP75 than the knockdown of the enhanced green fluorescent protein (EGFP). Furthermore, the total sugar content of the honeydew was lower when NcSP75 was knocked down. These results suggest that the NcSP75 protein contribute to successful and sustainable ingestion from the sieve elements of rice plants. The NcSP75 protein of N. cincticeps can, accordingly, be considered as a key effector for establishing compatible interaction with rice plants and could be a potential target for controlling this species.