An effective virus-based gene silencing method for functional genomics studies in common bean

Claudia Díaz-Camino, Padmanaban Annamalai, Federico Sanchez, Aardra Kachroo, Said A. Ghabrial

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

Background: Common bean (Phaseolus vulgaris L.) is a crop of economic and nutritious importance in many parts of the world. The lack of genomic resources have impeded the advancement of common bean genomics and thereby crop improvement. Although concerted efforts from the "Phaseomics" consortium have resulted in the development of several genomic resources, functional studies have continued to lag due to the recalcitrance of this crop for genetic transformation.Results: Here we describe the use of a bean pod mottle virus (BPMV)-based vector for silencing of endogenous genes in common bean as well as for protein expression. This BPMV-based vector was originally developed for use in soybean. It has been successfully employed for both protein expression and gene silencing in this species. We tested this vector for applications in common bean by targeting common bean genes encoding nodulin 22 and stearoyl-acyl carrier protein desaturase for silencing. Our results indicate that the BPMV vector can indeed be employed for reverse genetics studies of diverse biological processes in common bean. We also used the BPMV-based vector for expressing the green fluorescent protein (GFP) in common bean and demonstrate stable GFP expression in all common bean tissues where BPMV was detected.Conclusions: The availability of this vector is an important advance for the common bean research community not only because it provides a rapid means for functional studies in common bean, but also because it does so without generating genetically modified plants. Here we describe the detailed methodology and provide essential guidelines for the use of this vector for both gene silencing and protein expression in common bean. The entire VIGS procedure can be completed in 4-5 weeks.

Original languageEnglish
Article number16
JournalPlant Methods
Volume7
Issue number1
DOIs
StatePublished - Jun 13 2011

Bibliographical note

Funding Information:
This work was funded in part by grants from United Soybean Board and Kentucky soybean Promotion Board (to SAG and AK) and from DGAPA UNAM IN203807 and IN202009. We thank Xochitl Alvarado-Affantranger and Wendy Havens for their technical support and Amy Crume for greenhouse care.

Funding

This work was funded in part by grants from United Soybean Board and Kentucky soybean Promotion Board (to SAG and AK) and from DGAPA UNAM IN203807 and IN202009. We thank Xochitl Alvarado-Affantranger and Wendy Havens for their technical support and Amy Crume for greenhouse care.

FundersFunder number
Kentucky Soybean Promotion Board
United Soybean Board
Dirección General de Asuntos del Personal Académico, Universidad Nacional Autónoma de MéxicoIN202009, IN203807

    ASJC Scopus subject areas

    • Biotechnology
    • Genetics
    • Plant Science

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