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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 Nicotian...

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Autores principales: Dalakouras, Athanasios, Wassenegger, Michèle, McMillan, John N., Cardoza, Vinitha, Maegele, Ira, Dadami, Elena, Runne, Miriam, Krczal, Gabi, Wassenegger, Michael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5003833/
https://www.ncbi.nlm.nih.gov/pubmed/27625678
http://dx.doi.org/10.3389/fpls.2016.01327
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author Dalakouras, Athanasios
Wassenegger, Michèle
McMillan, John N.
Cardoza, Vinitha
Maegele, Ira
Dadami, Elena
Runne, Miriam
Krczal, Gabi
Wassenegger, Michael
author_facet Dalakouras, Athanasios
Wassenegger, Michèle
McMillan, John N.
Cardoza, Vinitha
Maegele, Ira
Dadami, Elena
Runne, Miriam
Krczal, Gabi
Wassenegger, Michael
author_sort Dalakouras, Athanasios
collection PubMed
description 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.
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spelling pubmed-50038332016-09-13 Induction of Silencing in Plants by High-Pressure Spraying of In vitro-Synthesized Small RNAs Dalakouras, Athanasios Wassenegger, Michèle McMillan, John N. Cardoza, Vinitha Maegele, Ira Dadami, Elena Runne, Miriam Krczal, Gabi Wassenegger, Michael Front Plant Sci Plant Science 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. Frontiers Media S.A. 2016-08-30 /pmc/articles/PMC5003833/ /pubmed/27625678 http://dx.doi.org/10.3389/fpls.2016.01327 Text en Copyright © 2016 Dalakouras, Wassenegger, McMillan, Cardoza, Maegele, Dadami, Runne, Krczal and Wassenegger. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Dalakouras, Athanasios
Wassenegger, Michèle
McMillan, John N.
Cardoza, Vinitha
Maegele, Ira
Dadami, Elena
Runne, Miriam
Krczal, Gabi
Wassenegger, Michael
Induction of Silencing in Plants by High-Pressure Spraying of In vitro-Synthesized Small RNAs
title Induction of Silencing in Plants by High-Pressure Spraying of In vitro-Synthesized Small RNAs
title_full Induction of Silencing in Plants by High-Pressure Spraying of In vitro-Synthesized Small RNAs
title_fullStr Induction of Silencing in Plants by High-Pressure Spraying of In vitro-Synthesized Small RNAs
title_full_unstemmed Induction of Silencing in Plants by High-Pressure Spraying of In vitro-Synthesized Small RNAs
title_short Induction of Silencing in Plants by High-Pressure Spraying of In vitro-Synthesized Small RNAs
title_sort induction of silencing in plants by high-pressure spraying of in vitro-synthesized small rnas
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5003833/
https://www.ncbi.nlm.nih.gov/pubmed/27625678
http://dx.doi.org/10.3389/fpls.2016.01327
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