Virus-mediated transient expression techniques enable genetic modification of Alopecurus myosuroides

A - Papers appearing in refereed journals

Mellado-Sanchez, M., Mcdiarmid, F., Cardoso, V., Kanyuka, K. and Macgregor, D. 2020. Virus-mediated transient expression techniques enable genetic modification of Alopecurus myosuroides. Plant Physiology. 183, pp. 455-459.

AuthorsMellado-Sanchez, M., Mcdiarmid, F., Cardoso, V., Kanyuka, K. and Macgregor, D.

Even though considerable progress has been made in weed ecology, weed molecular biology has been hindered by an inability to genetically manipulate weeds. Genetic manipulation is essential to demonstrate a causative relationship between genotype and phenotype. Herein we demonstrate that virus-mediated transient expression techniques developed for other monocots can be used in black-grass (Alopecurus myosuroides) for loss- and gain-of-function studies. We not only use virus induced gene silencing (VIGS) to create the black-grass exhibiting reduced PHYTOENE DESATURASE expression and virus-mediated overexpression (VOX) to drive GREEN FLUORESCENT PROTEIN, we demonstrate these techniques are applicable to testing hypotheses related to herbicide resistance in black-grass. We use VIGS to demonstrate that AmGSTF1 is necessary for the resistant biotype Peldon to survive fenoxaprop application and show the heterologous expression of the bialaphos resistance gene with VOX is sufficient to confer resistance to an otherwise lethal dose of glufosinate. Black-grass is the most problematic weed for winter-cereal farmers in the UK and Western Europe as it has rapidly evolved adaptions that allow it to effectively avoid current integrated weed management practices. Black-grass also reduces yields and therefore directly threatens food security and productivity. Novel disruptive technologies which mitigate resistance evolution and enable better control over this pernicious weed are therefore required. These virus-mediated protocols offer a step change in our ability to alter genes of interest under controlled laboratory conditions and therefore to gain a molecular-level understanding of how black-grass can survive in the agri-environment.

KeywordsBlack-grass; VIGS; VOX; AmGSTF1; Herbicide resistance; Bar; PDS
Year of Publication2020
JournalPlant Physiology
Journal citation183, pp. 455-459
Digital Object Identifier (DOI)
Web address (URL)
Open accessPublished as ‘gold’ (paid) open access
FunderBBSRC Industrial Strategy Challenge
Funder project or codeBBSRC Strategic Programme in Smart Crop Protection
Designing Future Wheat (DFW) [ISPG]
Publisher's version
Supplemental file
Output statusPublished
Publication dates
Online01 Apr 2020
Publication process dates
Accepted26 Mar 2020
PublisherAmerican Society of Plant Biologists (ASPB)

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