Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben Stadt Seeland, Germany.
Seed Science and Population Genetics, Institute of Plant Breeding, University of Hohenheim Stuttgart, Germany.
Front Plant Sci. 2015 Jan 20;5:770. doi: 10.3389/fpls.2014.00770. eCollection 2014.
Detailed and standardized protocols for plant cultivation in environmentally controlled conditions are an essential prerequisite to conduct reproducible experiments with precisely defined treatments. Setting up appropriate and well defined experimental procedures is thus crucial for the generation of solid evidence and indispensable for successful plant research. Non-invasive and high throughput (HT) phenotyping technologies offer the opportunity to monitor and quantify performance dynamics of several hundreds of plants at a time. Compared to small scale plant cultivations, HT systems have much higher demands, from a conceptual and a logistic point of view, on experimental design, as well as the actual plant cultivation conditions, and the image analysis and statistical methods for data evaluation. Furthermore, cultivation conditions need to be designed that elicit plant performance characteristics corresponding to those under natural conditions. This manuscript describes critical steps in the optimization of procedures for HT plant phenotyping systems. Starting with the model plant Arabidopsis, HT-compatible methods were tested, and optimized with regard to growth substrate, soil coverage, watering regime, experimental design (considering environmental inhomogeneities) in automated plant cultivation and imaging systems. As revealed by metabolite profiling, plant movement did not affect the plants' physiological status. Based on these results, procedures for maize HT cultivation and monitoring were established. Variation of maize vegetative growth in the HT phenotyping system did match well with that observed in the field. The presented results outline important issues to be considered in the design of HT phenotyping experiments for model and crop plants. It thereby provides guidelines for the setup of HT experimental procedures, which are required for the generation of reliable and reproducible data of phenotypic variation for a broad range of applications.
在环境可控条件下进行植物种植的详细和标准化方案是进行具有精确定义处理的可重复实验的必要前提。因此，建立适当和明确的实验程序对于产生可靠的证据至关重要，对于成功的植物研究也是不可或缺的。非侵入性和高通量（HT）表型分析技术提供了同时监测和量化数百株植物性能动态的机会。与小规模植物种植相比，HT 系统从概念和逻辑的角度对实验设计以及实际的植物种植条件、图像分析和统计数据分析方法都提出了更高的要求。此外，需要设计能够产生与自然条件下相应的植物性能特征的培养条件。本文描述了优化 HT 植物表型分析系统程序的关键步骤。从模式植物拟南芥开始，测试了 HT 兼容的方法，并针对生长基质、土壤覆盖、浇水制度、自动化植物种植和成像系统中的实验设计（考虑环境不均匀性）进行了优化。如代谢物分析结果所示，植物运动不会影响植物的生理状态。基于这些结果，建立了玉米 HT 种植和监测的程序。HT 表型分析系统中玉米营养生长的变化与田间观察到的变化非常吻合。所提出的结果概述了在设计用于模式植物和作物的 HT 表型分析实验时需要考虑的重要问题。这为建立 HT 实验程序提供了指导，这些程序对于产生广泛应用的表型变异可靠且可重复的数据是必需的。
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