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siRNA biogenesis and advances in topically applied dsRNA for controlling virus infections in tomato plants

A non-transgenic approach based on RNA interference was employed to induce protection against tomato mosaic virus (ToMV) infection in tomato plants. dsRNA molecules targeting the cp gene of ToMV were topically applied on plants prior to virus inoculation. Protection was dose-dependent and sequence-s...

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Autores principales: Rego-Machado, Camila M., Nakasu, Erich Y. T., Silva, João M. F., Lucinda, Natália, Nagata, Tatsuya, Inoue-Nagata, Alice K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7746768/
https://www.ncbi.nlm.nih.gov/pubmed/33335295
http://dx.doi.org/10.1038/s41598-020-79360-5
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author Rego-Machado, Camila M.
Nakasu, Erich Y. T.
Silva, João M. F.
Lucinda, Natália
Nagata, Tatsuya
Inoue-Nagata, Alice K.
author_facet Rego-Machado, Camila M.
Nakasu, Erich Y. T.
Silva, João M. F.
Lucinda, Natália
Nagata, Tatsuya
Inoue-Nagata, Alice K.
author_sort Rego-Machado, Camila M.
collection PubMed
description A non-transgenic approach based on RNA interference was employed to induce protection against tomato mosaic virus (ToMV) infection in tomato plants. dsRNA molecules targeting the cp gene of ToMV were topically applied on plants prior to virus inoculation. Protection was dose-dependent and sequence-specific. While no protection was achieved when 0–16 µg dsRNA were used, maximum rates of resistance (60 and 63%) were observed in doses of 200 and 400 µg/plant, respectively. Similar rates were also obtained against potato virus Y when targeting its cp gene. The protection was quickly activated upon dsRNA application and lasted for up to 4 days. In contrast, no detectable antiviral response was triggered by the dsRNA from a begomovirus genome, suggesting the method is not effective against phloem-limited DNA viruses. Deep sequencing was performed to analyze the biogenesis of siRNA populations. Although long-dsRNA remained in the treated leaves for at least 10 days, its systemic movement was not observed. Conversely, dsRNA-derived siRNA populations (mainly 21- and 22-nt) were detected in non-treated leaves, which indicates endogenous processing and transport through the plant. Altogether, this study provides critical information for the development of novel tools against plant viruses; strengths and limitations inherent to the systems are discussed.
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spelling pubmed-77467682020-12-18 siRNA biogenesis and advances in topically applied dsRNA for controlling virus infections in tomato plants Rego-Machado, Camila M. Nakasu, Erich Y. T. Silva, João M. F. Lucinda, Natália Nagata, Tatsuya Inoue-Nagata, Alice K. Sci Rep Article A non-transgenic approach based on RNA interference was employed to induce protection against tomato mosaic virus (ToMV) infection in tomato plants. dsRNA molecules targeting the cp gene of ToMV were topically applied on plants prior to virus inoculation. Protection was dose-dependent and sequence-specific. While no protection was achieved when 0–16 µg dsRNA were used, maximum rates of resistance (60 and 63%) were observed in doses of 200 and 400 µg/plant, respectively. Similar rates were also obtained against potato virus Y when targeting its cp gene. The protection was quickly activated upon dsRNA application and lasted for up to 4 days. In contrast, no detectable antiviral response was triggered by the dsRNA from a begomovirus genome, suggesting the method is not effective against phloem-limited DNA viruses. Deep sequencing was performed to analyze the biogenesis of siRNA populations. Although long-dsRNA remained in the treated leaves for at least 10 days, its systemic movement was not observed. Conversely, dsRNA-derived siRNA populations (mainly 21- and 22-nt) were detected in non-treated leaves, which indicates endogenous processing and transport through the plant. Altogether, this study provides critical information for the development of novel tools against plant viruses; strengths and limitations inherent to the systems are discussed. Nature Publishing Group UK 2020-12-17 /pmc/articles/PMC7746768/ /pubmed/33335295 http://dx.doi.org/10.1038/s41598-020-79360-5 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Rego-Machado, Camila M.
Nakasu, Erich Y. T.
Silva, João M. F.
Lucinda, Natália
Nagata, Tatsuya
Inoue-Nagata, Alice K.
siRNA biogenesis and advances in topically applied dsRNA for controlling virus infections in tomato plants
title siRNA biogenesis and advances in topically applied dsRNA for controlling virus infections in tomato plants
title_full siRNA biogenesis and advances in topically applied dsRNA for controlling virus infections in tomato plants
title_fullStr siRNA biogenesis and advances in topically applied dsRNA for controlling virus infections in tomato plants
title_full_unstemmed siRNA biogenesis and advances in topically applied dsRNA for controlling virus infections in tomato plants
title_short siRNA biogenesis and advances in topically applied dsRNA for controlling virus infections in tomato plants
title_sort sirna biogenesis and advances in topically applied dsrna for controlling virus infections in tomato plants
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7746768/
https://www.ncbi.nlm.nih.gov/pubmed/33335295
http://dx.doi.org/10.1038/s41598-020-79360-5
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