Sidhu Jagdeep Singh, Schneider Hannah M
Department of Plant Science, Pennsylvania State University, University Park, Pennsylvania 16802, USA.
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) OT Gatersleben, Seeland 06466, Germany
Cold Spring Harb Protoc. 2025 Sep 2;2025(9):pdb.prot108585. doi: 10.1101/pdb.prot108585.
Root anatomy plays a critical structural and functional role in the maize root system, and regulates edaphic stress tolerance. The function and genetic basis of several maize root anatomical traits for stress tolerance have been demonstrated. Leveraging root anatomical traits in maize thus holds great potential for developing cultivars with greater nutrient and water efficiency. Key for such approaches is the ability to characterize the root anatomy of plants of interest. Here, we outline a systematic method for preparing, imaging, and analyzing maize root cross-sections. The protocol describes root sectioning (by hand or using a vibratome), preparation of microscope slides and toluidine blue staining, imaging under a light microscope, and both manual and semiautomated methods for anatomical feature extraction from images. The protocol enables the visualization and quantification of various anatomical tissues and traits, and its simplicity, adaptability, and accessibility make it an ideal choice for both small- and large-scale phenotyping studies in maize and other plant species. This standardized protocol provides researchers with a comprehensive methodology to accurately dissect root structures, enabling in-depth analyses that are essential for understanding plant growth, development, and adaptive value for stress tolerance.
根系解剖结构在玉米根系系统中起着关键的结构和功能作用，并调节对土壤胁迫的耐受性。几种玉米根系解剖性状对胁迫耐受性的功能和遗传基础已得到证实。因此，利用玉米的根系解剖性状在培育具有更高养分和水分利用效率的品种方面具有巨大潜力。此类方法的关键在于能够对感兴趣的植物的根系解剖结构进行表征。在此，我们概述了一种用于制备、成像和分析玉米根横切面的系统方法。该方案描述了根切片（手动或使用振动切片机）、显微镜载玻片的制备和甲苯胺蓝染色、在光学显微镜下成像，以及从图像中提取解剖特征的手动和半自动方法。该方案能够实现对各种解剖组织和性状的可视化和量化，其简单性、适应性和可及性使其成为玉米和其他植物物种小规模和大规模表型研究的理想选择。这种标准化方案为研究人员提供了一种全面的方法来准确剖析根系结构，从而能够进行深入分析，这对于理解植物生长、发育以及对胁迫耐受性的适应价值至关重要。
