Rapid detection of potyviruses from crude plant extracts

Potyviruses (genus Potyvirus; family Potyviridae) are widely distributed and represent one of the most economically important genera of plant viruses. Therefore, their accurate detection is a key factor in developing efficient control strategies. However, this can sometimes be problematic particular...

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Autores principales: Silva, Gonçalo, Oyekanmi, Joshua, Nkere, Chukwuemeka K., Bömer, Moritz, Kumar, P. Lava, Seal, Susan E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5873530/
https://www.ncbi.nlm.nih.gov/pubmed/29378167
http://dx.doi.org/10.1016/j.ab.2018.01.019
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author Silva, Gonçalo
Oyekanmi, Joshua
Nkere, Chukwuemeka K.
Bömer, Moritz
Kumar, P. Lava
Seal, Susan E.
author_facet Silva, Gonçalo
Oyekanmi, Joshua
Nkere, Chukwuemeka K.
Bömer, Moritz
Kumar, P. Lava
Seal, Susan E.
author_sort Silva, Gonçalo
collection PubMed
description Potyviruses (genus Potyvirus; family Potyviridae) are widely distributed and represent one of the most economically important genera of plant viruses. Therefore, their accurate detection is a key factor in developing efficient control strategies. However, this can sometimes be problematic particularly in plant species containing high amounts of polysaccharides and polyphenols such as yam (Dioscorea spp.). Here, we report the development of a reliable, rapid and cost-effective detection method for the two most important potyviruses infecting yam based on reverse transcription-recombinase polymerase amplification (RT-RPA). The developed method, named ‘Direct RT-RPA’, detects each target virus directly from plant leaf extracts prepared with a simple and inexpensive extraction method avoiding laborious extraction of high-quality RNA. Direct RT-RPA enables the detection of virus-positive samples in under 30 min at a single low operation temperature (37 °C) without the need for any expensive instrumentation. The Direct RT-RPA tests constitute robust, accurate, sensitive and quick methods for detection of potyviruses from recalcitrant plant species. The minimal sample preparation requirements and the possibility of storing RPA reagents without cold chain storage, allow Direct RT-RPA to be adopted in minimally equipped laboratories and with potential use in plant clinic laboratories and seed certification facilities worldwide.
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spelling pubmed-58735302018-04-01 Rapid detection of potyviruses from crude plant extracts Silva, Gonçalo Oyekanmi, Joshua Nkere, Chukwuemeka K. Bömer, Moritz Kumar, P. Lava Seal, Susan E. Anal Biochem Article Potyviruses (genus Potyvirus; family Potyviridae) are widely distributed and represent one of the most economically important genera of plant viruses. Therefore, their accurate detection is a key factor in developing efficient control strategies. However, this can sometimes be problematic particularly in plant species containing high amounts of polysaccharides and polyphenols such as yam (Dioscorea spp.). Here, we report the development of a reliable, rapid and cost-effective detection method for the two most important potyviruses infecting yam based on reverse transcription-recombinase polymerase amplification (RT-RPA). The developed method, named ‘Direct RT-RPA’, detects each target virus directly from plant leaf extracts prepared with a simple and inexpensive extraction method avoiding laborious extraction of high-quality RNA. Direct RT-RPA enables the detection of virus-positive samples in under 30 min at a single low operation temperature (37 °C) without the need for any expensive instrumentation. The Direct RT-RPA tests constitute robust, accurate, sensitive and quick methods for detection of potyviruses from recalcitrant plant species. The minimal sample preparation requirements and the possibility of storing RPA reagents without cold chain storage, allow Direct RT-RPA to be adopted in minimally equipped laboratories and with potential use in plant clinic laboratories and seed certification facilities worldwide. Elsevier 2018-04-01 /pmc/articles/PMC5873530/ /pubmed/29378167 http://dx.doi.org/10.1016/j.ab.2018.01.019 Text en © 2018 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Silva, Gonçalo
Oyekanmi, Joshua
Nkere, Chukwuemeka K.
Bömer, Moritz
Kumar, P. Lava
Seal, Susan E.
Rapid detection of potyviruses from crude plant extracts
title Rapid detection of potyviruses from crude plant extracts
title_full Rapid detection of potyviruses from crude plant extracts
title_fullStr Rapid detection of potyviruses from crude plant extracts
title_full_unstemmed Rapid detection of potyviruses from crude plant extracts
title_short Rapid detection of potyviruses from crude plant extracts
title_sort rapid detection of potyviruses from crude plant extracts
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5873530/
https://www.ncbi.nlm.nih.gov/pubmed/29378167
http://dx.doi.org/10.1016/j.ab.2018.01.019
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