Graduate School of Advanced Technology and Science, Tokushima University, Tokushima, Japan.
Center for Collaboration among Agriculture, Industry, and Commerce, Tokushima University, Tokushima, Japan.
Sci Rep. 2017 Mar 30;7(1):507. doi: 10.1038/s41598-017-00501-4.
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.
园艺作物中的单性结实是一种重要的性状，具有农业价值，可用于各种工业用途以及直接的食用品质。在这里，我们展示了一种使用 CRISPR/Cas9 系统生成单性结实番茄植株的育种策略。我们优化了 CRISPR/Cas9 系统，将体细胞突变有效地引入到控制单性结实的关键基因 SlIAA9 中，在 T0 代的突变率高达 100%。此外，使用深度测序分析脱靶突变表明，我们定制的 gRNA 没有在宿主基因组中诱导额外的突变。再生突变体表现出叶片形状和无籽果实的形态变化——这是单性结实番茄的特征。分离的下一代（T1）也表现出与纯合突变基因组相关的严重表型。该系统可用于使用这种精确快速的育种技术，在各种品种的番茄以及其他主要园艺作物中生产单性结实番茄。
