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 RNAs(dsRNAs)fed to HV,and found that dsRNAs targetingβ-Actin messenger RNA(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 d.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.

BioClay^(TM)prolongs RNA interference-mediated crop protection against Botrytis cinerea

One of the most promising tools for the control of fungal plant diseases is spray-induced gene silencing(SIGS).In SIGS,small interfering RNA(siRNA)or double-stranded RNA(dsRNA)targeting essential or virulence-related pathogen genes are exogenously applied to plants and postharvest products to trigger RNA interference(RNAi)of the targeted genes,inhibiting fungal growth and disease.However,SIGS is limited by the unstable nature of RNA under environmental conditions.The use of layered double hydroxide or clay particles as carriers to deliver biologically active dsRNA,a formulation termed BioClay^(TM),can enhance RNA durability on plants,prolonging its activity against pathogens.Here,we demonstrate that dsRNA delivered as BioClay can prolong protection against Botrytis cinerea,a major plant fungal pathogen,on tomato leaves and fruit and on mature chickpea plants.BioClay increased the protection window from 1 to 3 weeks on tomato leaves and from 5 to 10 days on tomato fruits,when compared with naked dsRNA.In flowering chickpea plants,BioClay provided prolonged protection for up to 4 weeks,covering the critical period of poding,whereas naked dsRNA provided limited protection.This research represents a major step forward for the adoption of SIGS as an eco-friendly alternative to traditional fungicides.