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- Title
Editing net blotch susceptibility genes for disease resistance in barley using CRISPR/Cas9 through gene gun bombardment.
- Authors
Sung-Yong Kim; Brueggeman, Robert; Åhman, Inger; Li-Hua Zhu
- Abstract
Net blotch, a worldwide disease in barley (Hordeum vulgare), is caused by the necrotrophic fungal pathogen Pyrenophora teres f. teres ( Ptt), resulting in a destructive foliar disease with up to 40% yield losses in susceptible barley cultivars. Distinct Ptt isolates can trigger differential disease responses depending on barley genotype. The cultivar Rika is resistant to Ptt isolate 15A but susceptible to Ptt isolate 6A. Conversely, the cultivar Kombar is susceptible to Ptt isolate 15A and resistant to Ptt isolate 6A. The centromeric region of barley chromosome 6H contains dominant susceptibility to 15A, 6A and multiple other isolates and a candidate gene with allelic specificity has been cloned from each of Kombar (Spt.k) and Rika (Spt.r). Further analysis of these Spt1 alleles is necessary to confirm their function and eventually for exploring a novel resistance breeding strategy in barley based on knocking out genes for susceptibility to net blotch. The CRISPR (clustered, regularly interspaced, short palindromic repeats)/Cas (CRISPRassociated) technique has recently been developed for targeted gene editing. Various transformation methods can be used to deliver CRISPR/Cas-containing plasmids into plant cells or tissues. Among them, the Agrobacterium-mediated transformation method is commonly used, especially for dicots. In the case of spring barley, Agrobacterium-mediated transformation is only efficient with the cultivar Golden promise. In our study, we are establishing an efficient particle bombardment transformation protocol for barley using CRISPR/Cas9 constructs for gene knock-out utilizing spring barley genotypes other than Golden Promis. We are targeting the Spt alleles in Rika and Kombar to confirm their function for net blotch susceptibility.
- Subjects
PYRENOPHORA teres; BARLEY genetics; CRISPRS
- Publication
Cereal Research Communications, 2017, Vol 45, p29
- ISSN
0133-3720
- Publication type
Abstract
- DOI
10.1556/0806.45.2017.100