In the last years, tremendous progress has been made in the development of CRISPR/Cas-mediated genome editing tools. A number of natural CRISPR/Cas nuclease variants have been characterized. Engineered Cas proteins have been developed to minimize PAM restrictions, off-side effects and temperature sensitivity. Both kinds of enzymes have, by now, been applied widely and efficiently in many plant species to generate either single or multiple mutations at the desired loci by multiplexing. In addition to DSB-induced mutagenesis, specifically designed CRISPR/Cas systems allow more precise gene editing, resulting not only in random mutations but also in predefined changes. Applications in plants include gene targeting by homologous recombination, base editing and, more recently, prime editing. We will evaluate these different technologies for their prospects and practical applicability in plants. In addition, we will discuss a novel application of the Cas9 nuclease in plants, enabling the induction of heritable chromosomal rearrangements, such as inversions and translocations. This technique will make it possible to change genetic linkages in a programmed way and add another level of genome engineering to the toolbox of plant breeding. Also, strategies for tissue culture free genome editing were developed, which might be helpful to overcome the transformation bottlenecks in many crops. All in all, the recent advances of CRISPR/Cas technology will help agriculture to address the challenges of the twenty-first century related to global warming, pollution and the resulting food shortage.

近年来，CRISPR/Cas介导的基因组编辑工具的开发取得了巨大进展。许多天然 CRISPR/Cas 核酸酶变体已得到表征。工程化 Cas 蛋白的开发旨在最大限度地减少 PAM 限制、越位效应和温度敏感性。到目前为止，这两种酶已广泛有效地应用于许多植物物种，通过多重作用在所需位点产生单个或多个突变。除了 DSB 诱导的诱变之外，专门设计的 CRISPR/Cas 系统还可以实现更精确的基因编辑，不仅导致随机突变，还导致预定义的变化。在植物中的应用包括通过同源重组进行基因打靶、碱基编辑以及最近的引物编辑。我们将评估这些不同技术的前景和在植物中的实际适用性。此外，我们将讨论 Cas9 核酸酶在植物中的一种新应用，能够诱导可遗传的染色体重排，例如倒位和易位。这项技术将使以编程方式改变遗传联系成为可能，并为植物育种工具箱添加另一个水平的基因组工程。此外，还开发了无组织培养的基因组编辑策略，这可能有助于克服许多作物的转化瓶颈。总而言之，CRISPR/Cas技术的最新进展将帮助农业应对二十一世纪与全球变暖、污染和由此导致的粮食短缺有关的挑战。