Location: Weed and Insect Biology Research
Project Number: 3060-21220-033-023-T
Project Type: Trust Fund Cooperative Agreement
Start Date: Jul 1, 2022
End Date: Jun 30, 2026
Objective:
1) Complete our efforts to use CRISPR technology to knock out the VIP3 gene in Regina II to make it less sensitive to deacclimation. 2) Identify gene expression changes in canola and kochia when both plants are grown in proximity and at different densities under both field and greenhouse conditions. 3) Identify genes and processes that can be modified to enhance competitiveness and reduce yield loss in canola growing in the presence of weeds. 4) Complete phenotype the F3 recombinant inbred lines (RILs) families from the cross between the highly freezing tolerant winter genotype that was crossed to the freezing sensitive spring genotype for both flowering time and freezing tolerance. 5) Genotype F3 families and map freezing tolerance and flowering time loci.
Approach:
1) We will design guide RNAs to knock out this gene in the deacclimation sensitive variety ARS312. These constructs will target one or both of the VIP3 genes of canola will be engineered and transformed into canola varieties with rapid deacclimation rates. 2) RNAseq analysis will be run on root and leaf samples of kochia and canola already collected from each plot from objective 2. Additionally, RNAseq analysis will be run on leaf and root material collected from kochia and canola from green house grown plants where kochia a single kochia plant was surrounded by 2 to six canola plants or a single canola plant was surrounded by 4two to six kochia plants in a random complete block design with 6 reps. 4) Analysis of gene expression differences and gene set enrichment and network analysis will be used to identify regulators of kochia-canola interactions. 5) Continue work to complete study where twelve individuals each from our 250 independent F3 RILs populations derived from a cross between a freezing sensitive spring and a freezing resistant winter type canola will be grown to the 6 leaf stage, acclimated, and subjected to freezing stress. The plants will then be scored for freezing damage and an average freezing survival score for each line will be generated using AI technology. Likewise, the average days to flowering will also be scored for 12 individuals each from each RIL population both before and after vernalization.
