Loss of foreign dna inserts from barley stripe mosaic virus vectors- potential consequences for use in functional genomics studies

Authors: Scofield, Steven - Steve

Submitted to: International Journal of Plant Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/21/2023
Publication Date: 11/30/2023
Citation: Scofield, S.R. 2023. Loss of foreign dna inserts from barley stripe mosaic virus vectors- potential consequences for use in functional genomics studies. International Journal of Plant Biology. https://doi.org/10.3390/ijpb14040080.
DOI: https://doi.org/10.3390/ijpb14040080

Interpretive Summary: irus-induced gene silencing (VIGS) is a very useful technique to determine the function of plant genes. In VIGS, plants are infected with an engineered virus that causes a plant gene, chosen by the researcher, to be silenced or switched off. The function of the chosen gene can then be determined by observing how switching it off affects the plant. This report describes a new method for initiating VIGS in wheat in which seeds are infected instead of leaves. This a more simple method than currently used and results in more uniform gene silencing. The new technique will be helpful to wheat researchers and may accelerate the pace of wheat research.

Technical Abstract: Barley stripe mosaic virus (BSMV) has been adapted to be a useful tool for virus-induced gene silencing in wheat and other cereals. The current practice for BSMV VIGS is to inoculate leaves of cereal plants with either BSMV in vitro transcripts or viral particles. This method has proven to be successful, but is not without difficulties. Prime among these issues are that gene silencing is not established uniformly throughout the plant and only phenotypes manifested in leaves and other aerial plant organs can be examined. To alleviate these issues, a method involving inoculation of wheat seeds has been developed. In this study viral particles were prepared from Nicotiana benthamiana plants infected with a BSMV construct engineered to silence phytoene desaturase (PDS). It was found that incubation of wheat seeds with BSMV viral particles during seed imbibition resulted in more uniform VIGS than leaf inoculated plants. Seed inoculated planted showed increased areas of the photobleaching phenotype associated with PDS silencing, in comparison to leaf inoculated plants. This method promises to improve the utility of BSM VIGS for functional genomics studies in cereals.