Virus Induced Gene Silencing (VIGS) to test gene function in pulse crops
Lead Investigator(s)
Dr. Christopher Todd
Lead Investigator(s) Institution
University of Saskatchewan - Dept. of Biology
Objective
To develop improved VIGS vectors to reliably infect and suppress gene expression in common bean, fababean, chickpea, field peas and lentils; to improve protocols for introduction of VIGS constructs into pulse crops; to quantify the effectiveness of gene suppression in different tissues using diverse germplasm; to employ VIGS to silence candidate genes linked to traits of interest.
Outcome
The goal of this project was to develop a laboratory technique to transiently silence specific genes in a number of legume species for use in testing gene function. Direct inoculation of ALSV virus, or inoculation using a bacterial carrier was not effective on soybean, common bean or mung bean, however we developed a simple protocol that was effective in silencing the target gene in pea plants. We also developed a novel technique based on root regeneration to introduce ALSV vector into root tissue that showed 100% effectiveness in pea. Using this technique we were also able to demonstrate gene silencing in lentil with ALSV. Using this approach we were able to introduce a Bean Pod Mottle Virus into germinated soybean and common bean seedlings, but not ALSV, suggesting that specific interactions between ALSV and members of the bean family were responsible for the lack of gene silencing observed with other inoculation methods.
Genetics
SPG Contribution
$99,015.00
Project Status
Complete
Duration/Timeline of Project (Year to Year)
2014 - 2017
Co-funders
Saskatchewan Ministry of Agriculture - Agriculture Development Fund
Total Project Cost
$185,115.00