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Bean pod mottle virus: a new powerful tool for functional genomics studies in Pisum sativum

Pea (Pisum sativum L.) is an important legume worldwide. The importance of pea in arable rotations and nutritional value for both human and animal consumption have fostered sustained production and different studies to improve agronomic traits of interest. Moreover, complete sequencing of the pea ge...

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Autores principales: Meziadi, Chouaib, Blanchet, Sophie, Richard, Manon M.S., Pilet‐Nayel, Marie‐Laure, Geffroy, Valérie, Pflieger, Stéphanie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5067550/
https://www.ncbi.nlm.nih.gov/pubmed/26896301
http://dx.doi.org/10.1111/pbi.12537
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author Meziadi, Chouaib
Blanchet, Sophie
Richard, Manon M.S.
Pilet‐Nayel, Marie‐Laure
Geffroy, Valérie
Pflieger, Stéphanie
author_facet Meziadi, Chouaib
Blanchet, Sophie
Richard, Manon M.S.
Pilet‐Nayel, Marie‐Laure
Geffroy, Valérie
Pflieger, Stéphanie
author_sort Meziadi, Chouaib
collection PubMed
description Pea (Pisum sativum L.) is an important legume worldwide. The importance of pea in arable rotations and nutritional value for both human and animal consumption have fostered sustained production and different studies to improve agronomic traits of interest. Moreover, complete sequencing of the pea genome is currently underway and will lead to the identification of a large number of genes potentially associated with important agronomic traits. Because stable genetic transformation is laborious for pea, virus‐induced gene silencing (VIGS) appears as a powerful alternative technology for determining the function of unknown genes. In this work, we present a rapid and efficient viral inoculation method using DNA infectious plasmids of Bean pod mottle virus (BPMV)‐derived VIGS vector. Six pea genotypes with important genes controlling biotic and/or abiotic stresses were found susceptible to BPMV carrying a GFP reporter gene and showed fluorescence in both shoots and roots. In a second step, we investigated 37 additional pea genotypes and found that 30 were susceptible to BPMV and only 7 were resistant. The capacity of BPMV to induce silencing of endogenes was investigated in the most susceptible genotype using two visual reporter genes: PsPDS and PsKORRIGAN1 (PsKOR1) encoding PHYTOENE DESATURASE and a 1,4‐β‐D‐glucanase, respectively. The features of the ‘one‐step’ BPMV‐derived VIGS vector include (i) the ease of rub‐inoculation, without any need for biolistic or agro‐inoculation procedures, (ii) simple cost‐effective procedure and (iii) noninterference of viral symptoms with silencing. These features make BPMV the most adapted VIGS vector in pea to make low‐ to high‐throughput VIGS studies.
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spelling pubmed-50675502016-11-01 Bean pod mottle virus: a new powerful tool for functional genomics studies in Pisum sativum Meziadi, Chouaib Blanchet, Sophie Richard, Manon M.S. Pilet‐Nayel, Marie‐Laure Geffroy, Valérie Pflieger, Stéphanie Plant Biotechnol J Research Articles Pea (Pisum sativum L.) is an important legume worldwide. The importance of pea in arable rotations and nutritional value for both human and animal consumption have fostered sustained production and different studies to improve agronomic traits of interest. Moreover, complete sequencing of the pea genome is currently underway and will lead to the identification of a large number of genes potentially associated with important agronomic traits. Because stable genetic transformation is laborious for pea, virus‐induced gene silencing (VIGS) appears as a powerful alternative technology for determining the function of unknown genes. In this work, we present a rapid and efficient viral inoculation method using DNA infectious plasmids of Bean pod mottle virus (BPMV)‐derived VIGS vector. Six pea genotypes with important genes controlling biotic and/or abiotic stresses were found susceptible to BPMV carrying a GFP reporter gene and showed fluorescence in both shoots and roots. In a second step, we investigated 37 additional pea genotypes and found that 30 were susceptible to BPMV and only 7 were resistant. The capacity of BPMV to induce silencing of endogenes was investigated in the most susceptible genotype using two visual reporter genes: PsPDS and PsKORRIGAN1 (PsKOR1) encoding PHYTOENE DESATURASE and a 1,4‐β‐D‐glucanase, respectively. The features of the ‘one‐step’ BPMV‐derived VIGS vector include (i) the ease of rub‐inoculation, without any need for biolistic or agro‐inoculation procedures, (ii) simple cost‐effective procedure and (iii) noninterference of viral symptoms with silencing. These features make BPMV the most adapted VIGS vector in pea to make low‐ to high‐throughput VIGS studies. John Wiley and Sons Inc. 2016-02-20 2016-08 /pmc/articles/PMC5067550/ /pubmed/26896301 http://dx.doi.org/10.1111/pbi.12537 Text en © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Meziadi, Chouaib
Blanchet, Sophie
Richard, Manon M.S.
Pilet‐Nayel, Marie‐Laure
Geffroy, Valérie
Pflieger, Stéphanie
Bean pod mottle virus: a new powerful tool for functional genomics studies in Pisum sativum
title Bean pod mottle virus: a new powerful tool for functional genomics studies in Pisum sativum
title_full Bean pod mottle virus: a new powerful tool for functional genomics studies in Pisum sativum
title_fullStr Bean pod mottle virus: a new powerful tool for functional genomics studies in Pisum sativum
title_full_unstemmed Bean pod mottle virus: a new powerful tool for functional genomics studies in Pisum sativum
title_short Bean pod mottle virus: a new powerful tool for functional genomics studies in Pisum sativum
title_sort bean pod mottle virus: a new powerful tool for functional genomics studies in pisum sativum
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5067550/
https://www.ncbi.nlm.nih.gov/pubmed/26896301
http://dx.doi.org/10.1111/pbi.12537
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