The "one-step" Bean pod mottle virus (BPMV)-derived vector is a functional genomics tool for efficient overexpression of heterologous protein, virus-induced gene silencing and genetic mapping of BPMV R-gene in common bean (Phaseolus vulgaris L.)

BMC Plant Biology

Research Article

Manon MS Richard¹  Vincent Thareau¹  Céline Mazoyer¹  Fanny Mary¹  Sophie Blanchet¹  Chouaib Meziadi¹  Valérie Geffroy²  Stéphanie Pflieger³ 

[1] CNRS, Institut de Biologie des Plantes, UMR 8618, Université Paris Sud, Saclay Plant Sciences (SPS), 91405, Orsay, France;CNRS, Institut de Biologie des Plantes, UMR 8618, Université Paris Sud, Saclay Plant Sciences (SPS), 91405, Orsay, France;INRA, Unité Mixte de Recherche de Génétique Végétale, Université Paris Sud, IDEEV FR3284, Ferme du Moulon, 91190, Gif-sur-Yvette, France;CNRS, Institut de Biologie des Plantes, UMR 8618, Université Paris Sud, Saclay Plant Sciences (SPS), 91405, Orsay, France;Univ Paris Diderot, Sorbonne Paris Cité, 75205, Paris, France;

关键词: Disease resistance;    Functional validation;    Legume;    Phaseolus vulgaris;    RNAi;    Post-transcriptional gene silencing;    Soybean;    Virus resistance gene;   

DOI  :  10.1186/s12870-014-0232-4

 received in 2014-06-11, accepted in 2014-08-20,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundOver the last two years, considerable advances have been made in common bean (Phaseolus vulgaris L.) genomics, especially with the completion of the genome sequence and the availability of RNAseq data. However, as common bean is recalcitrant to stable genetic transformation, much work remains to be done for the development of functional genomics tools adapted to large-scale studies.ResultsHere we report the successful implementation of an efficient viral vector system for foreign gene expression, virus-induced gene silencing (VIGS) and genetic mapping of a BPMV resistance gene in common bean, using a "one-step" BPMV vector originally developed in soybean. With the goal of developing this vector for high-throughput VIGS studies in common bean, we optimized the conditions for rub-inoculation of infectious BPMV-derived plasmids in common bean cv. Black Valentine. We then tested the susceptibility to BPMV of six cultivars, and found that only Black Valentine and JaloEEP558 were susceptible to BPMV. We used a BPMV-GFP construct to detect the spatial and temporal infection patterns of BPMV in vegetative and reproductive tissues. VIGS of the PHYTOENE DESATURASE (PvPDS) marker gene was successfully achieved with recombinant BPMV vectors carrying fragments ranging from 132 to 391 bp. Finally, we mapped a gene for resistance to BPMV (R-BPMV) at one end of linkage group 2, in the vicinity of a locus (I locus) previously shown to be involved in virus resistance.ConclusionsThe "one-step" BPMV vector system therefore enables rapid and simple functional studies in common bean, and could be suitable for large-scale analyses. In the post-genomic era, these advances are timely for the common bean research community.

【 授权许可】

Unknown   
© Pflieger et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd. 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, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.