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Virus-Induced Gene Silencing pp 15–32Cite as

Rationale for Developing New Virus Vectors to Analyze Gene Function in Grasses Through Virus-Induced Gene Silencing

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 975))

Abstract

The exploding availability of genome and EST-based sequences from grasses requires a technology that allows rapid functional analysis of the multitude of genes that these resources provide. There are several techniques available to determine a gene’s function. For gene knockdown studies, silencing through RNAi is a powerful tool. Gene silencing can be accomplished through stable transformation or transient expression of a fragment of a target gene sequence. Stable transformation in rice, maize, and a few other species, although routine, remains a relatively low-throughput process. Transformation in other grass species is difficult and labor-intensive. Therefore, transient gene silencing methods including Agrobacterium-mediated and virus-induced gene silencing (VIGS) have great potential for researchers studying gene function in grasses. VIGS in grasses already has been used to determine the function of genes during pathogen challenge and plant development. It also can be used in moderate-throughput reverse genetics screens to determine gene function. However, the number of viruses modified to serve as silencing vectors in grasses is limited, and the silencing phenotype induced by these vectors is not optimal: the phenotype being transient and with moderate penetration throughout the tissue. Here, we review the most recent information available for VIGS in grasses and summarize the strengths and weaknesses in current virus–grass host systems. We describe ways to improve current virus vectors and the potential of other grass-infecting viruses for VIGS studies. This work is necessary because VIGS for the foreseeable future remains a higher throughput and more rapid system to evaluate gene function than stable transformation.

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Acknowledgments

The authors thank Kiran Mysore, Clemencia M. Rojas, and Muthappa Senthil-Kumar for critical review of the manuscript. This work was supported by The BioEnergy Science Center, which is a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science, and the Samuel Roberts Noble Foundation, Inc.

Note added in proof: A BSMV vector modified for both ligation-free cloning and expression behind a plant-active promoter was recently published: Yuan et al. 2011, PLoS ONE 6, e26468.

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  1. Plant Biology Division, The Samuel Roberts Noble Foundation Inc., Ardmore, OK, USA

    Hema Ramanna, Xin Shun Ding & Richard S. Nelson

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  1. Hema Ramanna
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  2. Xin Shun Ding
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Correspondence to Richard S. Nelson .

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  1. Institute of Botany, Justus-Liebig-University Gießen, Heinrich-Buff-Ring 38, Gießen, 35392, Germany

    Annette Becker

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Ramanna, H., Ding, X.S., Nelson, R.S. (2013). Rationale for Developing New Virus Vectors to Analyze Gene Function in Grasses Through Virus-Induced Gene Silencing. In: Becker, A. (eds) Virus-Induced Gene Silencing. Methods in Molecular Biology, vol 975. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-278-0_2

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