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ARS Home » Southeast Area » Dawson, Georgia » National Peanut Research Laboratory » Research » Publications at this Location » Publication #346656
Title: Detection of exogenous double-stranded RNA (dsRNA) movement in in vitro peanut plants

Author
item Faustinelli, Paola
item Power, Imana
item Arias De Ares, Renee

Submitted to: Plant Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/30/2018
Publication Date: 2/6/2018
Citation: Faustinelli, P.C., Power, I.L., Arias De Ares, R.S. 2018. Detection of exogenous double-stranded RNA (dsRNA) movement in in vitro peanut plants. Plant Biology. doi.org/10.1111/plb.12703.
DOI: https://doi.org/10.1111/plb.12703

Interpretive Summary: New technologies are needed to eliminate mycotoxins and/or fungal pathogens from agricultural products. RNA interference (RNAi) has shown potential to control fungi associated to crops. This is the first time that movement of exogenous DsiRNA in in vitro peanut plants was studied. The fact that the duplexes moved toward the pegs containing part of the aflatoxin-biosynthesis pathogen gene enables silencing aflatoxin production when Dicer-substrate RNAs (DsiRNA) comes into contact with and crosses to the Aspergillus cells growing on the seed. Developing new genomics tools can provide great advantages to researchers who work in the design of strategies to effectively and efficiently control pathogens in crop plants.

Technical Abstract: New technologies are needed to eliminate mycotoxins and/or fungal pathogens from agricultural products. RNA interference (RNAi) has shown potential to control fungi associated to crops. In RNAi, double-stranded RNA (dsRNA) targets homologous messenger RNA (mRNA) for cleavage, and can reach the mRNA of pathogens in close contact with the plant. The key element in this process is the movement of RNA signals cell-to-cell and over long distance within the plant, and between host plants and their parasites. In this study, we selected an important regulatory gene in the aflatoxin-biosynthesis pathway, aflS/aflR, necessary for the production of aflatoxins in Aspergillus spp. Based on its DNA sequence, we designed a Dicer-substrate RNAs (DsiRNA) to study duplex movement and persistence over time in in vitro peanut plant using stem-loop primers and Real Time-Polymerase Chain Reaction (RT-PCR) for DsiRNA detection. These preliminary results demonstrated that DsiRNA were absorbed and moved away from the point of application, spread systematically, and were transported rapidly probably through the phloem of the shoot to sink tissues, such as new auxiliary shoots, flowers and newly formed pegs. The DsiRNA remained detectable at least 30 days after treatment. This is the first time that movement of exogenous DsiRNA in in vitro peanut plants was studied. The fact that the duplexes moved toward the pegs containing part of the aflatoxin-biosynthesis pathogen gene enables silencing aflatoxin production when DsiRNA comes into contact with and crosses to the Aspergillus cells growing on the seed. The application of small RNAs could be a non-transformative option for mycotoxin contamination control.