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Virus-Induced Gene Silencing as a Scalable Tool to Study Drought Tolerance in Plants

  • Gavin M. George
  • Michael E. Ruckle
  • James R. Lloyd
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1287)

Abstract

Here we describe the methodology of using virus-induced gene silencing (VIGS) as a powerful and scalable tool to screen the function of genes that participate in adaptation to drought. Silencing of endogenous gene expression in Nicotiana benthamiana is achieved by systemic infection of the aerial parts of the plant with a virus engineered to contain homologous fragments of the target gene(s) of interest. Silenced plant material can be consistently produced with little optimization in less than 1 month without specialized equipment, using only simple cloning and transformation techniques. Although maximal silencing is localized to only a few leaves, when whole plants are subjected to water stress, the tissue from these silenced leaves can be characterized for physiological, biochemical, and transcriptional responses to determine the role of the candidate genes in drought tolerance.

Key words

Virus-induced gene silencing Drought response/s Reverse genetics 

Notes

Acknowledgements

We gratefully acknowledge the Swiss-South African Joint Research Program (Grant Number: S002533) for support in funding this project. In addition, we would like to thank Prof. Samuel C. Zeeman, Prof. Jens Kossmann, the Zürich-Basel Plant Science Centers’ Plant Fellows program, and the South African National Research Foundation for their continuing contributions and support.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Gavin M. George
    • 1
  • Michael E. Ruckle
    • 1
  • James R. Lloyd
    • 2
  1. 1.Department of BiologyETH ZurichZurichSwitzerland
  2. 2.Institute for Plant BiotechnologyUniversity of StellenboschStellenboschSouth Africa

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