Optimisation of tobacco rattle virus-induced gene silencing in Arabidopsis

Functional Plant Biology : FPB
Chang-Chun WangZhong Zheng

Abstract

Arabidopsis thaliana (L.) Heynh. is a model plant species in which to study plant gene functions. Recently developed virus-induced gene silencing (VIGS) offers a rapid and high-throughput technique platform for gene function analysis. In this paper we report optimisation of tobacco rattle virus (TRV)-induced gene silencing in Arabidopsis. The parameters potentially affecting the efficiency of VIGS in Arabidopsis were investigated. These included the concentration and pre-incubation of Agrobacterium inocula (agro-inocula), the concentration of acetosyringone included in agro-inocula, the Agrobacterium inoculation (agro-inoculation) method, the ecotypes and the growth stages of Arabidopsis plants for agro-inoculation, and the growth temperature of agro-inoculated plants. The optimised VIGS procedure involves preparing the agro-inocula with OD600 of 2.0, pre-incubating for 2 h in infiltration buffer containing 200 μm acetosyringone, agro-inoculating by vacuum infiltration, and growth of agro-inoculated plants at 22 -24°C. Following this procedure consistent and highly efficient VIGS was achieved for the genes encoding phytoene desaturase (PDS) and actin in Arabidopsis. The silencing phenotype lasts for at least 6 weeks, and is app...Continue Reading

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Citations

Jul 23, 2008·The Plant Journal : for Cell and Molecular Biology·Stéphanie PfliegerIsabelle Jupin
Oct 17, 2014·Journal of Integrative Plant Biology·Patrícia VidigalLuísa Carvalho
Sep 14, 2017·The Plant Journal : for Cell and Molecular Biology·Maria Ines ReyesDominique Robertson
Jul 26, 2015·Tree Physiology·Zedan ShenShaoliang Chen
Oct 17, 2007·Plant Cell Reports·Mandar R GodgePrakash P Kumar
May 24, 2013·International Journal of Molecular Sciences·Ying-Li ZhangZhen-Hui Gong
Aug 25, 2019·International Journal of Molecular Sciences·Hongqiu ZengHaitao Shi
Nov 8, 2015·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Mumtaz Ali SaandXin-Zhong Cai
Jan 28, 2014·Plant Molecular Biology Reporter·Shaun R Broderick, Michelle L Jones
Jul 4, 2006·Plant Physiology·Tessa M Burch-SmithS P Dinesh-Kumar
Jun 19, 2021·Applications in Plant Sciences·Shane CareyJocelyn C Hall

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Methods Mentioned

BETA
PCR

Software Mentioned

Quantity One
Origin
VIGS

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