Development of tobacco ringspot virus-based vectors for foreign gene expression and virus-induced gene silencing in a variety of plants

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Title
Development of tobacco ringspot virus-based vectors for foreign gene expression and virus-induced gene silencing in a variety of plants
Author(s)
F Zhao; Seungmo Lim; D Igori; Ran Hee Yoo; Suk Yoon KwonJae Sun Moon
Bibliographic Citation
Virology, vol. 492, pp. 166-178
Publication Year
2016
Abstract
We report here the development of tobacco ringspot virus (TRSV)-based vectors for the transient expression of foreign genes and for the analysis of endogenous gene function in plants using virus-induced gene silencing. The jellyfish green fluorescent protein (GFP) gene was inserted between the TRSV movement protein (MP) and coat protein (CP) regions, resulting in high in-frame expression of the RNA2-encoded viral polyprotein. GFP was released from the polyprotein via an N-terminal homologous MP-CP cleavage site and a C-terminal foot-and-mouth disease virus (FMDV) 2 A catalytic peptide in Nicotiana benthamiana. The VIGS target gene was introduced in the sense and antisense orientations into a SnaBI site, which was created by mutating the sequence following the CP stop codon. VIGS of phytoene desaturase (PDS) in N. benthamiana, Arabidopsis ecotype Col-0, cucurbits and legumes led to obvious photo-bleaching phenotypes. A significant reduction in PDS mRNA levels in silenced plants was confirmed by semi-quantitative RT-PCR.
Keyword
ArabidopsisCucurbitsForeign gene expressionFunctional genomicsLegumesNicotiana benthamianaPhytoene desaturaseTobacco ringspot virusVirus-induced gene silencing
ISSN
0042-6822
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.virol.2016.02.025
Type
Article
Appears in Collections:
Division of Biomaterials Research > 1. Journal Articles
Division of Biomaterials Research > Plant Systems Engineering Research > 1. Journal Articles
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