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Laura, Marina; Forti, Chiara; Barberini, Sara; Ciorba, Roberto; Mascarello, Carlo; Giovannini, Annalisa; Pistelli, Luisa; Pieracci, Ylenia; Lanteri, Anna Paola; Ronca, Agostina; Minuto, Andrea; Ruffoni, Barbara; Cardi, Teodoro; Savona, Marco
Plants, 06/2023, Volume: 12, Issue: 13Journal Article
(sweet basil) is an economically important aromatic herb; in Italy, approximately 1000 ha of "Genovese-type" basil are grown annually in greenhouses and open fields and are subjected to Downy Mildew (DM) disease, caused by , leading to huge crop losses. Mutation of the Susceptibility (S) gene DMR6 (Downy Mildew Resistant 6) has been proven to confer a broad-spectrum resistance to DM. In this work, an effective Genome Editing (GE) approach mediated by CRISPR/Cas9 in 'Italiko', the élite cultivar used to produce "Pesto Genovese D.O.P", was developed. A highly efficient genetic transformation method mediated by has been optimized from cotyledonary nodes, obtaining 82.2% of regenerated shoots, 84.6% of which resulted in Cas9+ plants. Eleven T0 lines presented different type of mutations in 60% were indel frameshift mutations with knock-out of of 'FT Italiko'. Analysis of six T1 transgene-free seedlings revealed that the mutations of T0 plants were inherited and segregated. Based on infection trials conducted on T0 plants, clone 22B showed a very low percentage of disease incidence after 14 days post infection. The aromatic profile of all in vitro edited plants was also reported; all of them showed oxygenated monoterpenes as the major fraction.
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