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

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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.

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Authors and Affiliations

  1. State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou, 310029, China

    Chang-jun Huang, Tong Zhang, Fang-fang Li, Xin-yue Zhang & Xue-ping Zhou

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  1. Chang-jun Huang
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  2. Tong Zhang
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  3. Fang-fang Li
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  4. Xin-yue Zhang
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Correspondence to Xue-ping Zhou.

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Project supported by the National Science and Technology Major Projects of China (No. 2009ZX08009-026B), the China Postdoctoral Science Foundation (No. 20090461375), and the National Basic Research Program (973) of China (No. 2006CB101903)

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Huang, Cj., Zhang, T., Li, Ff. et al. Development and application of an efficient virus-induced gene silencing system in Nicotiana tabacum using geminivirus alphasatellite. J. Zhejiang Univ. Sci. B 12, 83–92 (2011). https://doi.org/10.1631/jzus.B1000157

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