Open Access Honors Program Thesis
Tilahun Abebe, Honors Thesis Advisor, Department of Biology
Fusarium--Genetic engineering; CRISPR-associated protein 9;
Fusarium graminearum causes the fungal disease known as scab or head blight in cereals, including barley. There is no known resistance in barley against scab disease. Generating mutations in the fungus is a promising strategy to reduce infection. Research has shown that CRISPR/Cas9 is a very powerful tool for gene mutation. In this study, CRISPR/Cas9 vectors containing guide RNA sequences were constructed to mutate one visual marker gene (AUR1) and two genes essential for infection in F. graminearum (Tri5 and MGV1). Plasmids pFC332 and pFC334 were used to create vectors with the target genes using the USER cloning kit. The vectors were cloned into E. coli sand confirmed the presence of guide RNA sequences and accompanying sequences (promoter, terminator, and tracrRNA) using PCR and sequencing. The vectors will be used to mutagenize AUR2, Tri5, and MGV1 genes in F. graminearum in future studies. It is predicted that mutated F. graminearum would not be able to infect host barley (Hordeum vulgare) plants.
Year of Submission
Department of Biology
University Honors Designation
A thesis submitted in partial fulfillment of the requirements for the designation University Honors
1 PDF file (v, 25 pages)
©2019 Rebekah Akers
Akers, Rebekah, "Construction of CRISPR/Cas9 vectors for directed mutagenesis of Fusarium graminearum" (2019). Honors Program Theses. 368.