2015 article
Persistent Virus-Induced Gene Silencing in Asymptomatic Accessions of Arabidopsis
PLANT FUNCTIONAL GENOMICS: METHODS AND PROTOCOLS, 2ND EDITION, Vol. 1284, pp. 305–322.
MeSH headings : Arabidopsis / genetics; Arabidopsis / growth & development; Arabidopsis / virology; Begomovirus / physiology; Cloning, Molecular / methods; Gene Silencing; Gene Targeting / methods; Genetic Vectors / genetics; Host-Pathogen Interactions / genetics
TL;DR:
It is shown that genetic variation in the vast collection of Arabidopsis accessions can be leveraged to ameliorate viral symptomology that accompanies the VIGS procedure.
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Coupled with the advantages afforded by the model plant Arabidopsis, virus-induced gene silencing (VIGS) offers a rapid means to assess gene function. The geminivirus vector based on Cabbage leaf curl virus described here has the benefits of small insert size and persistent silencing of the target gene through the life cycle of the plant. Here, we show that genetic variation in the vast collection of Arabidopsis accessions can be leveraged to ameliorate viral symptomology that accompanies the VIGS procedure. The plasticity of phenotypes under different day lengths or temperature conditions can be exploited to achieve maximum silencing efficacy in either vegetative or inflorescence tissue, according to the question being asked. Protocols and vectors for Agro-infiltration of primary leaves, subapical pricking in older plants, and microprojectile bombardment are described.