Department of Biology, Western Kentucky University; Plant Biotechnology Division, CSIR Central Institute of Medicinal and Aromatic Plants;
Plant Biotechnology Division, CSIR Central Institute of Medicinal and Aromatic Plants; Academy of Scientific and Innovative Research (AcSIR).
J Vis Exp. 2023 Feb 10(192). doi: 10.3791/64876.
Comprehensive knowledge of plant root system architecture (RSA) development is critical for improving nutrient use efficiency and increasing crop cultivar tolerance to environmental challenges. An experimental protocol is presented for setting up the hydroponic system, plantlet growth, RSA spreading, and imaging. The approach used a magenta box-based hydroponic system containing polypropylene mesh supported by polycarbonate wedges. Experimental settings are exemplified by assessing the RSA of the plantlets under varying nutrient (phosphate [Pi]) supply. The system was established to examine the RSA of Arabidopsis, but it is readily adaptable to study other plants like Medicago sativa (Alfalfa). Arabidopsis thaliana (Col-0) plantlets are used in this investigation as an example to understand the plant RSA. Seeds are surface sterilized by treating ethanol and diluted commercial bleach, and kept at 4 °C for stratification. The seeds are germinated and grown on a liquid half-MS medium on a polypropylene mesh supported by polycarbonate wedges. The plantlets are grown under standard growth conditions for the desired number days, gently picked out from the mesh, and submersed in water-containing agar plates. Each root system of the plantlets is spread gently on the water-filled plate with the help of a round art brush. These Petri plates are photographed or scanned at high resolution to document the RSA traits. The root traits, such as primary root, lateral roots, and branching zone, are measured using the freely available ImageJ software. This study provides techniques for measuring plant root characteristics in controlled environmental settings. We discuss how to (1) grow the plantlets, and collect and spread root samples, (2) obtain pictures of spread RSA samples, (3) capture the images, and (4) use image analysis software to quantify root attributes. The advantage of the present method is the versatile, easy, and efficient measurement of the RSA traits.
全面了解植物根系结构（RSA）的发育对于提高养分利用效率和增加作物品种对环境挑战的耐受性至关重要。本文提出了一种水培系统设置、种苗生长、RSA 扩展和成像的实验方案。该方法使用基于品红色盒子的水培系统，其中包含由聚碳酸酯楔形物支撑的聚丙烯网。通过评估植物在不同养分（磷酸盐[Pi]）供应下的 RSA，举例说明了实验设置。该系统是为研究拟南芥的 RSA 而建立的，但它很容易适应研究其他植物，如紫花苜蓿（Alfalfa）。本研究以拟南芥（Col-0）种苗为例，用于理解植物 RSA。种子先用乙醇和稀释的商用漂白剂进行表面消毒，然后在 4°C 下进行分层。种子在聚丙烯网支撑的聚碳酸酯楔形物上的液体半 MS 培养基上发芽和生长。种苗在标准生长条件下生长所需的天数，然后轻轻从网中取出，浸入含琼脂的水中平板。用圆画刷帮助将种苗的每个根系轻轻展开在充满水的平板上。使用高分辨率拍摄或扫描这些 Petri 平板以记录 RSA 特征。使用免费的 ImageJ 软件测量种苗根系的特征，如主根、侧根和分枝区。本研究提供了在受控环境条件下测量植物根系特征的技术。我们讨论了如何（1）种植种苗，收集和展开根样本，（2）获得展开的 RSA 样本的图片，（3）捕获图像，以及（4）使用图像分析软件来量化根属性。本方法的优点是能够灵活、简便、高效地测量 RSA 特征。
