Robinson Hannah F, Vink Jochem N A
School of Biological Sciences, Victoria University of Wellington, Wellington, New Zealand.
Methods Mol Biol. 2025;2892:93-104. doi: 10.1007/978-1-0716-4330-3_7.
The detached leaf assay is a valuable method for studying plant-pathogen interactions, enabling the assessment of pathogenicity, plant resistance, and treatment effects. In this protocol, we outline how to set up a Phytophthora detached leaf assay and use non-expert machine learning tools to increase the reliability and throughput of the image analysis. Utilizing ilastik for pixel classification and Python scripts for segmentation, manual correction, and temporal linking, the pipeline provides objective and quantitative data over time. The protocol covers assay setup and image segmentation and outlines key considerations, providing a comprehensive guide for setting up and analyzing detached leaf assays. The very minimal material requirements and user-friendly software make this protocol accessible for all Phytophthora researchers.
离体叶片测定法是研究植物与病原体相互作用的一种有价值的方法，可用于评估致病性、植物抗性和处理效果。在本方案中，我们概述了如何建立疫霉离体叶片测定法，并使用非专业机器学习工具来提高图像分析的可靠性和通量。该流程利用ilastik进行像素分类，使用Python脚本进行分割、手动校正和时间链接，可随时间提供客观和定量的数据。该方案涵盖了测定设置和图像分割，并概述了关键注意事项，为建立和分析离体叶片测定提供了全面指南。极低的材料要求和用户友好的软件使该方案适用于所有疫霉研究人员。
