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
Abstract

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)https://plan.core-apps.com/pag_2019/abstract/37c3073a-1705-41b1-96b7-6ee573d390bb
Open accessPublished as bronze (free) open access
Web address (URL) of conference proceedingshttps://plan.core-apps.com/pag_2019/abstract/37c3073a-1705-41b1-96b7-6ee573d390bb
Funder project or codeDesigning Future Wheat (DFW) [ISPG]
FunderBiotechnology and Biological Sciences Research Council
Output statusPublished
Publication dates
Online2019
Publication process dates
Accepted2019
Copyright licenseCC BY

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