Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated 9 (CRISPR/Cas9) system has recently become one popular technology due to its efficiency, precision, and simplicity compared with other genome editing tools such as Zinc Finger Nucleases (ZFNs) and Transcription Activator Like Effector Nucleases (TALENs). Horticultural crops provide energy and health-keeping nutrients to humankind. Genome-editing technology has become widely adopted in horticultural breeding with the increasing demand for high yield and better-quality horticultural crops. Here, we describe the CRISPR/Cas9 system construction, its optimization, including sgRNA promoter, sgRNA design, Cas9 protein promoter, SpCas9 variants and orthologs, and vector delivery methods. We also summarized the application of this technology in horticultural plants for stress responses enhancement, fruit quality improvement, and cultivation traits modification. This detailed review was compiled to help establish comprehensive understanding of the CRISPR/Cas9 systems and provide a reference for further developing this technology to manipulate horticultural plant traits effectively.

与锌指核酸酶 (ZFN) 和转录激活因子样效应器等其他基因组编辑工具相比，Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR 相关 9 (CRISPR/Cas9) 系统由于其效率、精度和简单性而最近成为一种流行的技术核酸酶 (TALEN)。园艺作物为人类提供能量和保健营养。随着人们对高产优质园艺作物的需求不断增加，基因组编辑技术在园艺育种中得到广泛应用。在这里，我们描述了 CRISPR/Cas9 系统的构建及其优化，包括 sgRNA 启动子、sgRNA 设计、Cas9 蛋白启动子、SpCas9 变体和直系同源物以及载体传递方法。我们还总结了该技术在园艺植物中用于增强应激反应、改善果实品质和改变栽培性状的应用。编写此详细综述旨在帮助建立对 CRISPR/Cas9 系统的全面了解，并为进一步开发该技术以有效操纵园艺植物性状提供参考。