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Induction of silencing in plants by high-pressure spraying of in vitro-synthesized small RNAs

In this report we describe a method for the delivery of small interfering RNAs (siRNAs) into plant cells. In vitro synthesized siRNAs that were designed to target the coding region of a GREEN FLUORESCENT PROTEIN (GFP) transgene were applied by various methods onto GFP-expressing transgenic Nicotiana...

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Main Authors: Dalakouras, Athanasios (Author) , Wassenegger, Michèle (Author) , McMillan, John N. (Author) , Cardoza, Vinitha (Author) , Maegele, Ira (Author) , Dadami, Elena (Author) , Runne, Miriam (Author) , Krczal, Gabi (Author) , Wassenegger, Michael (Author)
Format: Article (Journal)
Language:English
Published: 30 August 2016
In: Frontiers in plant science
Year: 2016, Volume: 7
ISSN:1664-462X
DOI:10.3389/fpls.2016.01327
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.3389/fpls.2016.01327
Verlag, lizenzpflichtig, Volltext: https://www.frontiersin.org/articles/10.3389/fpls.2016.01327/full
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Author Notes:Athanasios Dalakouras, Michèle Wassenegger, John N. McMillan, Vinitha Cardoza, Ira Maegele, Elena Dadami, Miriam Runne, Gabi Krczal and Michael Wassenegger
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Summary:In this report we describe a method for the delivery of small interfering RNAs (siRNAs) into plant cells. In vitro synthesized siRNAs that were designed to target the coding region of a GREEN FLUORESCENT PROTEIN (GFP) transgene were applied by various methods onto GFP-expressing transgenic Nicotiana benthamiana plants to trigger RNA silencing. In contrast to mere siRNA applications, including spraying, syringe injection and infiltration of siRNAs that all failed to induce RNA silencing, high pressure spraying of siRNAs resulted in efficient local and systemic silencing of the GFP transgene, with comparable efficiency as was achieved with biolistic siRNA introduction. High-pressure spraying of siRNAs with sizes of 21, 22 and 24 nucleotides (nt) led to local GFP silencing. Small RNA deep sequencing revealed that no shearing of siRNAs was detectable by high-pressure spraying. Systemic silencing was basically detected upon spraying of 22 nt siRNAs. Local and systemic silencing developed faster and more extensively upon targeting the apical meristem than spraying of mature leaves.
Item Description:Gesehen am 06.05.2020
Physical Description:Online Resource
ISSN:1664-462X
DOI:10.3389/fpls.2016.01327

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