The involvement of systemic RNA interference deficient-1-like (SIL1) in cellular dsRNA uptake in Acyrthosiphon pisum

Systemic RNA interference deficient-1-like(SIL1)is considered a core component in dsRNA uptake in some insect species.Investigation related to the potential function of SIL1 in dsRNA uptake can contribute to a further understanding of RNA interference(RNAi)mechanisms in insects and agricultural pest control.However,the role of SIL1 in dsRNA uptake in insects such as aphids remains controversial.We have thoroughly analyzed the role of SIL1 from the model aphid Acyrthosiphon pisum(ApSIL1)in cellular dsRNA to clarify its function.First,the induced expression of ApSIL1 upon dsRNA oral exposure provided a vital clue for the possible involvement of ApSIL1 in cellular dsRNA uptake.Subsequent in vivo experiments using the RNAi-of-RNAi approach for ApSIL1 supported our hypothesis that the silencing efficiencies of reporter genes were reduced after inhibition of ApSIL1 expression.The impaired biological phenotypes of aphids,including cumulative average offspring,deformities of the nymph,and mortality upon pathogen infection,were then observed in the treatment group.Thereafter,in vitro dual-luciferase reporter assay showed compelling evidence that the luciferin signal was significantly attenuated when dsluciferase or dsGFP was transferred into ApSIL1-transfected Drosophila S2 cells.These observations further confirmed that the signal of Cy3-labeled dsRNA was rapidly attenuated with time in ApSIL1-transfected Drosophila S2 cells.Overall,these findings conclusively establish that ApSIL1 is involved in dsRNA uptake in A.pisum.