Silencing of aphid genes by dsRNA feeding from plants.

<h4>Background</h4>RNA interference (RNAi) is a valuable reverse genetics tool to study gene function in various organisms, including hemipteran insects such as aphids. Previous work has shown that RNAi-mediated knockdown of pea aphid (Acyrthosiphon pisum) genes can be achieved through d...

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Main Authors: Marco Pitino, Alexander D Coleman, Massimo E Maffei, Christopher J Ridout, Saskia A Hogenhout
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2011-01-01
Series:PLoS ONE
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21998682/?tool=EBI
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spelling doaj-acfbd69233544762a8187212d16728bf2021-03-04T01:29:56ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-01610e2570910.1371/journal.pone.0025709Silencing of aphid genes by dsRNA feeding from plants.Marco PitinoAlexander D ColemanMassimo E MaffeiChristopher J RidoutSaskia A Hogenhout<h4>Background</h4>RNA interference (RNAi) is a valuable reverse genetics tool to study gene function in various organisms, including hemipteran insects such as aphids. Previous work has shown that RNAi-mediated knockdown of pea aphid (Acyrthosiphon pisum) genes can be achieved through direct injection of double-stranded RNA (dsRNA) or small-interfering RNAs (siRNA) into the pea aphid hemolymph or by feeding these insects on artificial diets containing the small RNAs.<h4>Methodology/principal findings</h4>In this study, we have developed the plant-mediated RNAi technology for aphids to allow for gene silencing in the aphid natural environment and minimize handling of these insects during experiments. The green peach aphid M. persicae was selected because it has a broad plant host range that includes the model plants Nicotiana benthamiana and Arabidopsis thaliana for which transgenic materials can relatively quickly be generated. We targeted M. persicae Rack1, which is predominantly expressed in the gut, and M. persicae C002 (MpC002), which is predominantly expressed in the salivary glands. The aphids were fed on N. benthamiana leaf disks transiently producing dsRNA corresponding to these genes and on A. thaliana plants stably producing the dsRNAs. MpC002 and Rack-1 expression were knocked down by up to 60% on transgenic N. benthamiana and A. thaliana. Moreover, silenced M. persicae produced less progeny consistent with these genes having essential functions.<h4>Conclusions/significance</h4>Similar levels of gene silencing were achieved in our plant-mediated RNAi approach and published silencing methods for aphids. Furthermore, the N. benthamiana leaf disk assay can be developed into a screen to assess which genes are essential for aphid survival on plants. Our results also demonstrate the feasibility of the plant-mediated RNAi approach for aphid control.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21998682/?tool=EBI
collection DOAJ
language English
format Article
sources DOAJ
author Marco Pitino
Alexander D Coleman
Massimo E Maffei
Christopher J Ridout
Saskia A Hogenhout
spellingShingle Marco Pitino
Alexander D Coleman
Massimo E Maffei
Christopher J Ridout
Saskia A Hogenhout
Silencing of aphid genes by dsRNA feeding from plants.
PLoS ONE
author_facet Marco Pitino
Alexander D Coleman
Massimo E Maffei
Christopher J Ridout
Saskia A Hogenhout
author_sort Marco Pitino
title Silencing of aphid genes by dsRNA feeding from plants.
title_short Silencing of aphid genes by dsRNA feeding from plants.
title_full Silencing of aphid genes by dsRNA feeding from plants.
title_fullStr Silencing of aphid genes by dsRNA feeding from plants.
title_full_unstemmed Silencing of aphid genes by dsRNA feeding from plants.
title_sort silencing of aphid genes by dsrna feeding from plants.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2011-01-01
description <h4>Background</h4>RNA interference (RNAi) is a valuable reverse genetics tool to study gene function in various organisms, including hemipteran insects such as aphids. Previous work has shown that RNAi-mediated knockdown of pea aphid (Acyrthosiphon pisum) genes can be achieved through direct injection of double-stranded RNA (dsRNA) or small-interfering RNAs (siRNA) into the pea aphid hemolymph or by feeding these insects on artificial diets containing the small RNAs.<h4>Methodology/principal findings</h4>In this study, we have developed the plant-mediated RNAi technology for aphids to allow for gene silencing in the aphid natural environment and minimize handling of these insects during experiments. The green peach aphid M. persicae was selected because it has a broad plant host range that includes the model plants Nicotiana benthamiana and Arabidopsis thaliana for which transgenic materials can relatively quickly be generated. We targeted M. persicae Rack1, which is predominantly expressed in the gut, and M. persicae C002 (MpC002), which is predominantly expressed in the salivary glands. The aphids were fed on N. benthamiana leaf disks transiently producing dsRNA corresponding to these genes and on A. thaliana plants stably producing the dsRNAs. MpC002 and Rack-1 expression were knocked down by up to 60% on transgenic N. benthamiana and A. thaliana. Moreover, silenced M. persicae produced less progeny consistent with these genes having essential functions.<h4>Conclusions/significance</h4>Similar levels of gene silencing were achieved in our plant-mediated RNAi approach and published silencing methods for aphids. Furthermore, the N. benthamiana leaf disk assay can be developed into a screen to assess which genes are essential for aphid survival on plants. Our results also demonstrate the feasibility of the plant-mediated RNAi approach for aphid control.
url https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21998682/?tool=EBI
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