Highly Efficient CRISPR/Cas9 Mediated Gene Editing in Ocimum basilicum ‘FT Italiko’ to Induce Resistance to Peronospora belbahrii

Autor: Marina Laura, Chiara Forti, Sara Barberini, Roberto Ciorba, Carlo Mascarello, Annalisa Giovannini, Luisa Pistelli, Ylenia Pieracci, Anna Paola Lanteri, Agostina Ronca, Andrea Minuto, Barbara Ruffoni, Teodoro Cardi, Marco Savona
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: Plants, Vol 12, Iss 13, p 2395 (2023)
Druh dokumentu: article
ISSN: 2223-7747
DOI: 10.3390/plants12132395
Popis: Ocimum basilicum (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 Peronospora belbahrii, 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 O. basilicum ‘Italiko’, the élite cultivar used to produce “Pesto Genovese D.O.P”, was developed. A highly efficient genetic transformation method mediated by A. tumefaciens 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 ObDMR6; 60% were indel frameshift mutations with knock-out of ObDMR6 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.
Databáze: Directory of Open Access Journals