title: Rapid breeding of parthenocarpic tomato plants using CRISPR/Cas9.
abstract: Parthenocarpy in horticultural crop plants is an important trait with agricultural value for various industrial purposes as well as direct eating quality. Here, we demonstrate a breeding strategy to generate parthenocarpic tomato plants using the CRISPR/Cas9 system. We optimized the CRISPR/Cas9 system to introduce somatic mutations effectively into SlIAA9-a key gene controlling parthenocarpy-with mutation rates of up to 100% in the T0 generation. Furthermore, analysis of off-target mutations using deep sequencing indicated that our customized gRNAs induced no additional mutations in the host genome. Regenerated mutants exhibited morphological changes in leaf shape and seedless fruit-a characteristic of parthenocarpic tomato. And the segregated next generation (T1) also showed a severe phenotype associated with the homozygous mutated genome. The system developed here could be applied to produce parthenocarpic tomato in a wide variety of cultivars, as well as other major horticultural crops, using this precise and rapid breeding technique.
authors.affiliation: Graduate School of Advanced Technology and Science, Tokushima University, Tokushima, Japan.
authors.forename: Risa
authors.initials: R
authors.lastname: Ueta
authors.affiliation: Graduate School of Advanced Technology and Science, Tokushima University, Tokushima, Japan.
authors.forename: Chihiro
authors.initials: C
authors.lastname: Abe
authors.affiliation: Center for Collaboration among Agriculture, Industry, and Commerce, Tokushima University, Tokushima, Japan.
authors.forename: Takahito
authors.initials: T
authors.lastname: Watanabe
authors.affiliation: Center for Collaboration among Agriculture, Industry, and Commerce, Tokushima University, Tokushima, Japan.
authors.forename: Shigeo S
authors.initials: SS
authors.lastname: Sugano
authors.affiliation: Graduate School of Advanced Technology and Science, Tokushima University, Tokushima, Japan.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Ishihara
authors.affiliation: Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ezura
authors.affiliation: Faculty of Bioscience and Bioindustry, Tokushima University, Tokushima, Japan.
authors.forename: Yuriko
authors.initials: Y
authors.lastname: Osakabe
authors.affiliation: Faculty of Bioscience and Bioindustry, Tokushima University, Tokushima, Japan. kosakabe@tokushima-u.ac.jp.
authors.forename: Keishi
authors.identifier: http://orcid.org/0000-0002-6227-6913
authors.initials: K
authors.lastname: Osakabe
mesh_terms: D001483:Base Sequence; D001947:Breeding; D064113:CRISPR-Cas Systems / Q000235:genetics*; D055786:Gene Knockout Techniques; D018551:Solanum lycopersicum / Q000235:genetics* / Q000254:growth & development*; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics; D010312:Parthenogenesis / Q000235:genetics*; D010641:Phenotype; D030821:Plants, Genetically Modified; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems