CRISPR/Cas9-Targeted Knockout of Rice Susceptibility Genes OsDjA2 and OsERF104 Reveals Alternative Sources of Resistance to Pyricularia oryzae
Fabiano T.P.K. Távora,
Anne Cécile Meunier,
Aurore Vernet,
Murielle Portefaix,
Joëlle Milazzo,
Henri Adreit,
Didier Tharreau,
Octávio L. Franco,
Angela Mehta
Affiliations
Fabiano T.P.K. Távora
Department of Genetics and Biotechnology, Federal University of Juiz de Fora, Juiz de Fora-MG 36036330, Brazil; Embrapa Genetic Resources and Biotechnology, Brasília-DF 70770-917, Brazil
Anne Cécile Meunier
Genetic Improvement and Adaptation of Mediterranean and Tropical Plants, Agricultural Research Center for International Development (CIRAD), Montpellier 34398 CEDEX 5, France; CIRAD-INRAe-Agro Institute, University of Montpellier, Montpellier 34000, France
Aurore Vernet
Genetic Improvement and Adaptation of Mediterranean and Tropical Plants, Agricultural Research Center for International Development (CIRAD), Montpellier 34398 CEDEX 5, France; CIRAD-INRAe-Agro Institute, University of Montpellier, Montpellier 34000, France
Murielle Portefaix
Genetic Improvement and Adaptation of Mediterranean and Tropical Plants, Agricultural Research Center for International Development (CIRAD), Montpellier 34398 CEDEX 5, France; CIRAD-INRAe-Agro Institute, University of Montpellier, Montpellier 34000, France
Joëlle Milazzo
Plant Health Institute of Montpellier, CIRAD, Montpellier 34398, France; University of Montpellier, National Institute of Agronomic Research, Research Institute for Development, Montpellier SupAgro, Montpellier 34398, France
Henri Adreit
Plant Health Institute of Montpellier, CIRAD, Montpellier 34398, France; University of Montpellier, National Institute of Agronomic Research, Research Institute for Development, Montpellier SupAgro, Montpellier 34398, France
Didier Tharreau
Plant Health Institute of Montpellier, CIRAD, Montpellier 34398, France; University of Montpellier, National Institute of Agronomic Research, Research Institute for Development, Montpellier SupAgro, Montpellier 34398, France
Octávio L. Franco
Department of Genomics Science and Biotechnology, Proteomics Analysis Center, Catholic University of Brasília, Brasília-DF 71966-900, Brazil; S-Inova Biotech, Catholic University of Dom Bosco, Campo Grande-MS 79117-900, Brazil
Angela Mehta
Embrapa Genetic Resources and Biotechnology, Brasília-DF 70770-917, Brazil; Corresponding author:
Rice genes OsDjA2 and OsERF104, encoding a chaperone protein and an APETELA2/ ethylene-responsive factor, respectively, are strongly induced in a compatible interaction with blast fungus, and also have function in plant susceptibility validated through gene silencing. Here, we reported the CRISPR/Cas9 knockout of OsDjA2 and OsERF104 genes resulting in considerable improvement of blast resistance. A total of 15 OsDjA2 (62.5%) and 17 OsERF104 (70.8%) T0 transformed lines were identified from 24 regenerated plants for each target and used in downstream experiments. Phenotyping of homozygous T1 mutant lines revealed not only a significant decrease in the number of blast lesions but also a reduction in the percentage of diseased leaf area, compared with the infected control plants. Our results supported CRISPR/Cas9-mediated target mutation in rice susceptibility genes as a potential and alternative breeding strategy for building resistance to blast disease.
