Abstract
The pea aphid, Acyrthosiphon pisum, is an important agricultural pest and biological model organism, and RNA interference (RNAi) is an important tool for functional genomics and for insect pest management. However, the efficiency of RNAi in pea aphids is variable, limiting its application in aphids. In this study, we present optimized conditions for inducing and increasing the gene silencing efficiency of RNAi in pea aphids. The optimal gene silencing of the target Aphunchback gene was achieved by injecting 600 ng double-stranded (ds) RNA, and the highest mRNA depletion rate (74%) was detected at 36 h after injection. Moreover, the same gene silencing conditions were used to achieve transcript silencing for nine different genes in the pea aphid, although the silencing efficiencies for the different genes varied. Furthermore, the pre-exposure of aphids to dsRNA (600 ng dsGFP) led to significant hunchback silencing following a secondary exposure to 60 ng of dshunchback, a dose which did not lead to gene silencing when independently injected. The information presented here can be exploited to develop more efficient RNAi bioassays for pea aphids, both as gene functional study tools and an insect pest control strategy.
Highlights
Over the past two decades, RNA interference (RNAi) has become a powerful tool for gene functional studies in various organisms by delivering gene-specific double-stranded RNA, and it is recognized as a next-generation insect pest control tool (Wang et al, 2017; Zhang et al, 2017)
To explore whether an increase in the transcript levels of ApDcr2, ApR2d2, and ApAgo2 could lead to an increase in the RNAi efficiency, we evaluated the effects of pre-injecting double-stranded RNA (dsRNA) into adult pea aphids later verifying the gene silencing efficiency following a second injection
We provided a general image of dose- and time-dependent gene silencing efficiencies in pea aphids
Summary
Over the past two decades, RNA interference (RNAi) has become a powerful tool for gene functional studies in various organisms by delivering gene-specific double-stranded RNA (dsRNA), and it is recognized as a next-generation insect pest control tool (Wang et al, 2017; Zhang et al, 2017). In insects, both viral infections (Marques et al, 2013; Niu et al, 2016) and non-specific exogenous dsRNAs (Flenniken and Andino, 2013; Cappelle et al, 2016) can induce the activity of the small interfering (si) RNA pathway. Some factors that may influence the efficacy of RNAi efficacy include dsRNA degradation (Singh et al, 2017), dsRNA uptake (Huvenne and Smagghe, 2010), the expression of RNAi-related genes (Garbutt and Reynolds, 2012), the transcript abundance of target genes (Scott et al, 2013) and viral infections (Swevers et al, 2013)
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