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- Title
Simultaneous CRISPR/Cas9 Editing of Three PPO Genes Reduces Fruit Flesh Browning in Solanum melongena L.
- Authors
Maioli, Alex; Gianoglio, Silvia; Moglia, Andrea; Acquadro, Alberto; Valentino, Danila; Milani, Anna Maria; Prohens, Jaime; Orzaez, Diego; Granell, Antonio; Lanteri, Sergio; Comino, Cinzia
- Abstract
Polyphenol oxidases (PPOs) catalyze the oxidization of polyphenols, which in turn causes the browning of the eggplant berry flesh after cutting. This has a negative impact on fruit quality for both industrial transformation and fresh consumption. Ten PPO genes (named SmelPPO1 - 10) were identified in eggplant thanks to the recent availability of a high-quality genome sequence. A CRISPR/Cas9-based mutagenesis approach was applied to knock-out three target PPO genes (SmelPPO4, SmelPPO5 , and SmelPPO6) , which showed high transcript levels in the fruit after cutting. An optimized transformation protocol for eggplant cotyledons was used to obtain plants in which Cas9 is directed to a conserved region shared by the three PPO genes. The successful editing of the SmelPPO4, SmelPPO5 , and SmelPPO6 loci of in vitro regenerated plantlets was confirmed by Illumina deep sequencing of amplicons of the target sites. Besides, deep sequencing of amplicons of the potential off-target loci identified in silico proved the absence of detectable non-specific mutations. The induced mutations were stably inherited in the T1 and T2 progeny and were associated with a reduced PPO activity and browning of the berry flesh after cutting. Our results provide the first example of the use of the CRISPR/Cas9 system in eggplant for biotechnological applications and open the way to the development of eggplant genotypes with low flesh browning which maintain a high polyphenol content in the berries.
- Subjects
EGGPLANT; CRISPRS; FRUIT; FRUIT quality; GENES; MUTAGENESIS
- Publication
Frontiers in Plant Science, 2020, Vol 11, pN.PAG
- ISSN
1664-462X
- Publication type
Article
- DOI
10.3389/fpls.2020.607161