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CLC number: Q93

On-line Access: 2011-02-04

Received: 2010-04-28

Revision Accepted: 2010-11-29

Crosschecked: 2011-01-12

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Journal of Zhejiang University SCIENCE B 2011 Vol.12 No.2 P.83-92

http://doi.org/10.1631/jzus.B1000157


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


Author(s):  Chang-jun Huang, Tong Zhang, Fang-fang Li, Xin-yue Zhang, Xue-ping Zhou

Affiliation(s):  State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China

Corresponding email(s):   zzhou@zju.edu.cn

Key Words:  Virus induced gene silencing, Geminivirus, Alphasatellite, Nicotiana tabacum


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Chang-jun Huang, Tong Zhang, Fang-fang Li, Xin-yue Zhang, Xue-ping Zhou. Development and application of an efficient virus-induced gene silencing system in Nicotiana tabacum using geminivirus alphasatellite[J]. Journal of Zhejiang University Science B, 2011, 12(2): 83-92.

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author="Chang-jun Huang, Tong Zhang, Fang-fang Li, Xin-yue Zhang, Xue-ping Zhou",
journal="Journal of Zhejiang University Science B",
volume="12",
number="2",
pages="83-92",
year="2011",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1000157"
}

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%T Development and application of an efficient virus-induced gene silencing system in Nicotiana tabacum using geminivirus alphasatellite
%A Chang-jun Huang
%A Tong Zhang
%A Fang-fang Li
%A Xin-yue Zhang
%A Xue-ping Zhou
%J Journal of Zhejiang University SCIENCE B
%V 12
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%P 83-92
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%D 2011
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1000157

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T1 - Development and application of an efficient virus-induced gene silencing system in Nicotiana tabacum using geminivirus alphasatellite
A1 - Chang-jun Huang
A1 - Tong Zhang
A1 - Fang-fang Li
A1 - Xin-yue Zhang
A1 - Xue-ping Zhou
J0 - Journal of Zhejiang University Science B
VL - 12
IS - 2
SP - 83
EP - 92
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PB - Zhejiang University Press & Springer
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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.

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