Zahid Muhammad Awais, Kieu Nam Phuong
Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Lomma, Sweden.
Curr Protoc. 2025 Jul;5(7):e70180. doi: 10.1002/cpz1.70180.
Potato (Solanum tuberosum L.) yield is highly sensitive to drought stress, yet robust phenotyping methods for drought tolerance remain scarce. To address this challenge, we present a rapid, high-throughput hydroponic assay designed as an efficient pre-screening tool for evaluating potato cultivars and CRISPR-edited lines. This protocol serves as a foundational screen, enabling researchers to identify the most promising genotypes before committing resources to more extensive soil or field trials. The system uses repurposed pipette tip boxes as low-cost, scalable hydroponic units, making the method highly accessible. Over a five-week period, plantlets are subjected to controlled 24-hr osmotic stress with polyethylene glycol (PEG-6000) and then assessed for resilience based on biomass, photosynthetic measurements, and visual recovery. This resource-efficient assay provides a controlled environment to minimize experimental noise and has been successfully applied to characterize CRISPR-edited potato lines. By providing a reproducible platform for initial evaluation, this protocol accelerates the selection pipeline for developing robust potato varieties for a changing climate. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Preparation of stem explants and in vitro rooting Basic Protocol 2: Assembly and maintenance of tip-box hydroponic units Basic Protocol 3: PEG-induced drought treatment and recovery Basic Protocol 4: Post-stress phenotypic, biomass, and photosynthetic analysis.
马铃薯（Solanum tuberosum L.）产量对干旱胁迫高度敏感，但用于耐旱性的强大表型分析方法仍然稀缺。为应对这一挑战，我们提出了一种快速、高通量的水培试验方法，设计为一种高效的预筛选工具，用于评估马铃薯品种和经CRISPR编辑的品系。该方案作为一种基础筛选方法，使研究人员能够在投入资源进行更广泛的土壤或田间试验之前，识别出最有前景的基因型。该系统使用重新利用的移液器吸头盒作为低成本、可扩展的水培单元，使该方法易于使用。在五周的时间里，将幼苗置于用聚乙二醇（PEG - 6000）控制的24小时渗透胁迫下，然后根据生物量、光合测量和视觉恢复情况评估其恢复力。这种资源高效的试验提供了一个可控环境，以尽量减少实验噪声，并已成功应用于表征经CRISPR编辑的马铃薯品系。通过提供一个可重复的初始评估平台，该方案加快了为适应气候变化培育健壮马铃薯品种的选择流程。© 2025作者。由Wiley Periodicals LLC出版的《Current Protocols》。基本方案1：茎外植体的制备和离体生根 基本方案2：吸头盒水培单元的组装和维护 基本方案3：PEG诱导的干旱处理和恢复 基本方案4：胁迫后表型、生物量和光合分析。
