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Journal Article Research Support, Non-U.S. Gov't Ppublish 13 citations

PubMed: 36221782 DOI: 10.1111/pbi.13948

Novel resistance to the Bymovirus BaMMV established by targeted mutagenesis of the PDIL5-1 susceptibility gene in barley.

Plant Biotechnol J 21(2):331-341

Authors: Robert Eric Hoffie, Dragan Perovic, Antje Habekuß, Frank Ordon, Jochen Kumlehn

Affiliations

  • Plant Reproductive Biology, Leibniz-Institute of Plant Genetics and Crop Plant Research (IPK), Seeland, Germany.
  • Institute for Resistance Research and Stress Tolerance, Julius Kuehn Institute (JKI), Federal Research Centre for Cultivated Plants, Quedlinburg, Germany.

Abstract

The Potyviridae are the largest family of plant-pathogenic viruses. Members of this family are the soil-borne bymoviruses barley yellow mosaic virus (BaYMV) and barley mild mosaic virus (BaMMV), which, upon infection of young winter barley seedlings in autumn, can cause yield losses as high as 50%. Resistance breeding plays a major role in coping with these pathogens. However, some viral strains have overcome the most widely used resistance. Thus, there is a need for novel sources of resistance. In ancient landraces and wild relatives of cultivated barley, alleles of the susceptibility factor PROTEIN DISULFIDE ISOMERASE LIKE 5-1 (PDIL5-1) were identified to confer resistance to all known strains of BaYMV and BaMMV. Although the gene is highly conserved throughout all eukaryotes, barley is thus far the only species for which PDIL5-1-based virus resistance has been reported. Whereas introgression by crossing to the European winter barley breeding pool is tedious, time-consuming and additionally associated with unwanted linkage drag, the present study exemplifies an approach to targeted mutagenesis of two barley cultivars employing CRISPR-associated endonuclease technology to induce site-directed mutations similar to those described for PDIL5-1 alleles that render certain landraces resistant. Homozygous primary mutants were produced in winter barley, and transgene-free homozygous M mutants were produced in spring barley. A variety of mutants carrying novel PDIL5-1 alleles were mechanically inoculated with BaMMV, by which all frameshift mutations and certain in-frame mutations were demonstrated to confer resistance to this virus. Under greenhouse conditions, virus-resistant mutants showed no adverse effects in terms of growth and yield.

MeSH

Mutagenesis, Infections, Biotechnology, Clustered Regularly Interspaced Short Palindromic Repeats, Mutation, Genes, Breeding, Alleles, Mosaic Viruses, Protein Disulfide-Isomerases

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