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RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)

RNA interference (RNAi), a gene-silencing mechanism that involves providing double-stranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become e...

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Autores principales: Camargo, Roberto A., Barbosa, Guilherme O., Possignolo, Isabella Presotto, Peres, Lazaro E. P., Lam, Eric, Lima, Joni E., Figueira, Antonio, Marques-Souza, Henrique
Formato: Online Artículo Texto
Lenguaje:English
Publicado: PeerJ Inc. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5162399/
https://www.ncbi.nlm.nih.gov/pubmed/27994959
http://dx.doi.org/10.7717/peerj.2673
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author Camargo, Roberto A.
Barbosa, Guilherme O.
Possignolo, Isabella Presotto
Peres, Lazaro E. P.
Lam, Eric
Lima, Joni E.
Figueira, Antonio
Marques-Souza, Henrique
author_facet Camargo, Roberto A.
Barbosa, Guilherme O.
Possignolo, Isabella Presotto
Peres, Lazaro E. P.
Lam, Eric
Lima, Joni E.
Figueira, Antonio
Marques-Souza, Henrique
author_sort Camargo, Roberto A.
collection PubMed
description RNA interference (RNAi), a gene-silencing mechanism that involves providing double-stranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become evident. The production of transgenic plants expressing dsRNA molecules that target essential insect genes could provide a means of specific gene silencing in larvae that feed on these plants, resulting in larval phenotypes that range from loss of appetite to death. In this report, we show that the tomato leafminer (Tuta absoluta), a major threat to commercial tomato production, can be targeted by RNAi. We selected two target genes (Vacuolar ATPase-A and Arginine kinase) based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet for T. absoluta, we used two approaches to deliver dsRNA into tomato leaflets. The first approach was based on the uptake of dsRNA by leaflets and the second was based on “in planta-induced transient gene silencing” (PITGS), a well-established method for silencing plant genes, used here for the first time to deliver in planta-transcribed dsRNA to target insect genes. Tuta absoluta larvae that fed on leaves containing dsRNA of the target genes showed an ∼60% reduction in target gene transcript accumulation, an increase in larval mortality and less leaf damage. We then generated transgenic ‘Micro-Tom’ tomato plants that expressed hairpin sequences for both genes and observed a reduction in foliar damage by T. absoluta in these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest.
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spelling pubmed-51623992016-12-19 RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum) Camargo, Roberto A. Barbosa, Guilherme O. Possignolo, Isabella Presotto Peres, Lazaro E. P. Lam, Eric Lima, Joni E. Figueira, Antonio Marques-Souza, Henrique PeerJ Agricultural Science RNA interference (RNAi), a gene-silencing mechanism that involves providing double-stranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become evident. The production of transgenic plants expressing dsRNA molecules that target essential insect genes could provide a means of specific gene silencing in larvae that feed on these plants, resulting in larval phenotypes that range from loss of appetite to death. In this report, we show that the tomato leafminer (Tuta absoluta), a major threat to commercial tomato production, can be targeted by RNAi. We selected two target genes (Vacuolar ATPase-A and Arginine kinase) based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet for T. absoluta, we used two approaches to deliver dsRNA into tomato leaflets. The first approach was based on the uptake of dsRNA by leaflets and the second was based on “in planta-induced transient gene silencing” (PITGS), a well-established method for silencing plant genes, used here for the first time to deliver in planta-transcribed dsRNA to target insect genes. Tuta absoluta larvae that fed on leaves containing dsRNA of the target genes showed an ∼60% reduction in target gene transcript accumulation, an increase in larval mortality and less leaf damage. We then generated transgenic ‘Micro-Tom’ tomato plants that expressed hairpin sequences for both genes and observed a reduction in foliar damage by T. absoluta in these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest. PeerJ Inc. 2016-12-15 /pmc/articles/PMC5162399/ /pubmed/27994959 http://dx.doi.org/10.7717/peerj.2673 Text en ©2016 Camargo et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.
spellingShingle Agricultural Science
Camargo, Roberto A.
Barbosa, Guilherme O.
Possignolo, Isabella Presotto
Peres, Lazaro E. P.
Lam, Eric
Lima, Joni E.
Figueira, Antonio
Marques-Souza, Henrique
RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)
title RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)
title_full RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)
title_fullStr RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)
title_full_unstemmed RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)
title_short RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)
title_sort rna interference as a gene silencing tool to control tuta absoluta in tomato (solanum lycopersicum)
topic Agricultural Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5162399/
https://www.ncbi.nlm.nih.gov/pubmed/27994959
http://dx.doi.org/10.7717/peerj.2673
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