Carlsen Frida Meijer, Westberg Ida, Johansen Ida Elisabeth, Andreasson Erik, Petersen Bent Larsen
Section for Plant Glycobiology, Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg C, Denmark.
Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Lomma, Sweden.
CRISPR J. 2025 Feb;8(1):37-50. doi: 10.1089/crispr.2024.0068. Epub 2024 Dec 4.
The potato family includes a highly diverse cultivar repertoire and has a high potential for nutritional yield improvement and refinement but must in line with other crops be adapted to biotic and abiotic stresses, for example, accelerated by climate change and environmental demands. The combination of pluripotency, high ploidy, and relative ease of protoplast isolation, transformation, and regeneration together with clonal propagation through tubers makes potato highly suitable for precise genetic engineering. Most potato varieties are tetraploid having a very high prevalence of length polymorphisms and small nucleotide polymorphisms between alleles, often complicating CRISPR-Cas editing designs and strategies. CRISPR-Cas editing in potato can be divided into (i) characterization of target area and -aided editing design, (ii) isolation and editing of protoplast cells, and (iii) the subsequent explant regeneration from single protoplast cells. Implementation of efficient CRISPR-Cas editing relies on efficient editing at the protoplast (cell pool) level and on robust high-throughput editing scoring methods at the cell pool and explant level. Gene and chromatin structure are additional features to optionally consider. Strategies and solutions for addressing key steps in genome editing of potato, including light conditions and schemes for reduced exposure to hormones during explant regeneration, which is often linked to somaclonal variation, are highlighted.
茄科植物包含高度多样的栽培品种库，在提高营养产量和品质方面具有很大潜力，但与其他作物一样，必须适应生物和非生物胁迫，例如因气候变化和环境要求而加剧的胁迫。多能性、高倍性以及原生质体分离、转化和再生相对容易，再加上通过块茎进行克隆繁殖，使得马铃薯非常适合进行精确的基因工程。大多数马铃薯品种是四倍体，等位基因之间长度多态性和单核苷酸多态性的发生率很高，这常常使CRISPR-Cas编辑设计和策略变得复杂。马铃薯中的CRISPR-Cas编辑可分为：（i）目标区域的表征和辅助编辑设计；（ii）原生质体细胞的分离和编辑；（iii）随后从单个原生质体细胞再生外植体。高效CRISPR-Cas编辑的实施依赖于原生质体（细胞库）水平的高效编辑以及细胞库和外植体水平强大的高通量编辑评分方法。基因和染色质结构是另外需要酌情考虑的特征。文中强调了马铃薯基因组编辑关键步骤的策略和解决方案，包括光照条件以及外植体再生过程中减少激素暴露的方案，外植体再生通常与体细胞无性系变异有关。
