Consumption of miRNA-Mediated Insect-Resistant Transgenic Rice Pollen Does Not Harm Apis mellifera Adults

RNAi-based insect-resistant genetically engineered (IRGE) crops represent a promising approach for pest management by suppressing gene expressions or translation. A developed microRNA-mediated IRGE rice line expressing endogenous Chilo suppressalis Csu-novel-260 shows significant resistance to target pests. The nontarget insect Apis mellifera is an important pollinator used as a surrogate species for the ecological risk assessment of IRGE plants. To simulate a worst-case scenario, the full-length C. suppressalis and A. mellifera disembodied (dib) cDNAs were cloned. The dib 3'-untranslated regions shared 58.06% nucleotide sequence similarity between C. suppressalis and A. mellifera. No potential Csu-novel-260 binding site in Amdib was detected through the bioinformatics analysis. A dietary RNAi toxicity assay of the impacts of ingested Csu-novel-260 on A. mellifera adults showed that the survival rates of RNAi-treated A. mellifera did not significantly differ from those in the blank control (CK) and negative control (NC) treatments. The Csu-novel-260 uptake by A. mellifera peaked at 8 days postfeeding and then gradually decreased. The Amdib expression was not affected by the RNAi assay days or treatments. These results suggest that A. mellifera adults are not susceptible to high doses of Csu-novel-260 in the dietary RNAi assay and that the impact of miRNA-mediated IRGE plants on A. mellifera is negligible.