Journal of Zhejiang University SCIENCE B

, Volume 12, Issue 2, pp 83–92

Development and application of an efficient virus-induced gene silencing system in Nicotiana tabacum using geminivirus alphasatellite

Authors

  • Chang-jun Huang
    • State Key Laboratory of Rice Biology, Institute of BiotechnologyZhejiang University
  • Tong Zhang
    • State Key Laboratory of Rice Biology, Institute of BiotechnologyZhejiang University
  • Fang-fang Li
    • State Key Laboratory of Rice Biology, Institute of BiotechnologyZhejiang University
  • Xin-yue Zhang
    • State Key Laboratory of Rice Biology, Institute of BiotechnologyZhejiang University
    • State Key Laboratory of Rice Biology, Institute of BiotechnologyZhejiang University
Article

DOI: 10.1631/jzus.B1000157

Cite this article as:
Huang, C., Zhang, T., Li, F. et al. J. Zhejiang Univ. Sci. B (2011) 12: 83. doi:10.1631/jzus.B1000157

Abstract

Virus-induced gene silencing (VIGS) is a recently developed technique for characterizing the function of plant genes by gene transcript suppression and is increasingly used to generate transient loss-of-function assays. Here we report that the 2mDNA1, a geminivirus satellite vector, can induce efficient gene silencing in Nicotiana tabacum with Tobacco curly shoot virus. We have successfully silenced the β-glucuronidase (GUS) gene in GUS transgenic N. tabacum plants and the sulphur desaturase (Su) gene in five different N. tabacum cultivars. These pronounced and severe silencing phenotypes are persistent and ubiquitous. Once initiated in seedlings, the silencing phenotype lasted for the entire life span of the plants and silencing could be induced in a variety of tissues and organs including leaf, shoot, stem, root, and flower, and achieved at any growth stage. This system works well between 18–32 °C. We also silenced the NtEDS1 gene and demonstrated that NtEDS1 is essential for N gene mediated resistance against Tobacco mosaic virus in N. tabacum. The above results indicate that this system has great potential as a versatile VIGS system for routine functional analysis of genes in N. tabacum.

Key words

Virus induced gene silencingGeminivirusAlphasatelliteNicotiana tabacum

CLC number

Q93

Copyright information

© Zhejiang University and Springer-Verlag Berlin Heidelberg 2011