Location: Small Grains and Potato Germplasm Research
Project Number: 2050-21000-034-022-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Aug 1, 2018
End Date: Jan 31, 2021
Objective:
To use gene editing approaches to genetically engineer Fusarium Head Blight (FHB) resistance in barley. Our specific objectives for this two-year project are: (1) construction of barley HvHSK, Hv2OGO and HvEIN2 transient and integrating CRISPR-editing vectors; (2) production of HvHSK-, Hv2OGO- and HvEIN2-edited barley plants (cv. Conlon); (3) evaluation of HvHSK, Hv2OGO and HvEIN2 mutant barley plants for FHB resistance; (4) editing the HvUGT promoter in the FHB susceptible barley cv. Morex to study the HvUGT gene expression kinetics; (5) editing the Nud gene in two-rowed and hulled Conlon and ND Genesis barley to study the effect of hull type on the FHB development; (6) editing the Vrs1 gene in the six-rowed cultivar Morex to study the effect of row type on the FHB development; and (7) developing barley anther culture for CRISPR-gene editing and barley engineering.
Approach:
Engineer and test resistance to FHB in barley through gene editing technology and develop resistant two-row and six-row barley varieties through breeding. Existing methods for transformation of embryogenic callus cultures will be used and modified as necessary for success with cultivars other than Golden Promise that are relevant to North American commercial production. Genetic modifications will include knockouts of genes associated with FHB susceptibility in Arabidopsis or wheat, and editing of other genes and promoters with association to FHB symptoms, accomplished via the construction and introduction of vectors (gRNA and Cas9) designed for transient expression. The development of a suitable anther culture system for barley will involve optimization of culture conditions and genotype. To facilitate success, collaboration will be established with researchers in the University of Minnesota and Oregon State University.
