Knocking down wheat meiotic genes through Virus-induced gene-silencing (VIGS)

C2 - Non-edited contributions to conferences

Desjardins, S., Kanyuka, K. and Higgins, J. D. 2019. Knocking down wheat meiotic genes through Virus-induced gene-silencing (VIGS). The Plant and Animal Genome XXVII Conference (PAG). San Diego, CA, USA 12 Jan 2019

AuthorsDesjardins, S., Kanyuka, K. and Higgins, J. D.
TypeC2 - Non-edited contributions to conferences

Virus-induced gene-silencing (VIGS) is a reverse genetic technology that can be used to assess gene function in model and crop species. The technique exploits a natural viral defence response in plants, whereby invading viral RNAs are detected, and this triggers a targeted post-transcriptional gene silencing (PTGS) mechanism. In VIGS, a small fragment of a plant gene of interest is inserted into a viral vector to generate a recombinant virus that, upon infection, leads to silencing of both the foreign viral RNA and the endogenous target gene; creating knockdown lines.
We have applied barley stripe mosaic virus (BSMV)-mediated VIGS to the study of recombination in hexaploid wheat, and have optimised the targeting of meiotic genes at peak expression. Following VIGS, successful down-regulation of gene expression can be assessed by qRT-PCR, and meiotic phenotypes analysed cytologically. An initial proof-of-concept study was conducted by silencing the recombinase TaDMC1, which resulted in a severe meiotic phenotype with the majority of homologous chromosomes remaining unpaired, visible as univalents, and we are currently expanding this approach to incorporate additional targets. VIGS is proving to be a rapid and cost-effective way of assessing the function of meiotic genes in wheat, enabling us to efficiently screen a broad range of candidate genes, so that further analyses using TILLING or CRISPR/Cas may be employed on promising targets.

KeywordsVIGS; Wheat; Meiosis
Year of Publication2019
Conference locationSan Diego, CA, USA
Event date12 Jan 2019
16 Jan 2019
Web address (URL)
Open accessPublished as bronze (free) open access
Web address (URL) of conference proceedings
Funder project or codeDesigning Future Wheat (DFW) [ISPG]
FunderBiotechnology and Biological Sciences Research Council
Output statusPublished
Publication dates
Publication process dates
Copyright licenseCC BY

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