Complete protection from Henosepilachna vigintioctopunctata by expressing long double-stranded RNAs in potato plastids

RNA interference (RNAi) has emerged as a powerful technology for pest management. Previously, we have shown that plastid-mediated RNAi (PM-RNAi) can be utilized to control the Colorado potato beetle, an insect pest in the Chrysomelidae family; however, whether this technology is suitable for controlling pests in the Coccinellidae remained unknown. The coccinellid 28-spotted potato ladybird (Henosepilachna vigintioctopunctata; HV) is a serious pest of solanaceous crops. In this study, we identified three efficient target genes (β-Actin, SRP54, and SNAP) for RNAi using in vitro double-stranded RNA (dsRNA) fed to HV, and found that dsRNAs targeting β-Actin mRNA (dsACT) induced more potent RNAi than those targeting the other two genes. We next generated transplastomic and nuclear transgenic potato (Solanum tuberosum) plants expressing HV dsACT. Long dsACT stably accumulated to up to 0.7% of the total cellular RNA in the transplastomic plants, at least three orders of magnitude higher than in the nuclear transgenic plants. Notably, the transplastomic plants also exhibited a significantly stronger resistance to HV, killing all larvae within 6 days. Our data demonstrate the potential of PM-RNAi as an efficient pest control measure for HV, extending the application range of this technology to Coccinellidae pests. This article is protected by copyright. All rights reserved.