Pandey Bipin K, Atkinson Jonathan A, Sturrock Craig J
Division of Plant and Crop Science, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, UK.
The Hounsfield Facility, Division of Agricultural and Environmental Sciences, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, UK.
Bio Protoc. 2021 Dec 20;11(24):e4252. doi: 10.21769/BioProtoc.4252.
Roots are the prime organ for nutrient and water uptake and are therefore fundamental to the growth and development of plants. However, physical challenges of a heterogeneous environment and diverse edaphic stresses affect root growth in soil. Compacted soil is a serious global problem, causing inhibition of root elongation, which reduces surface area and impacts resource foraging. Visualisation and quantification of roots in soil is difficult due to this growth substrate's opaque nature; however, non-destructive imaging technologies are now becoming more widely available to plant and soil scientists working to address this challenge. We have recently developed an integrated approach, combining X-ray Computed Tomography (X-ray CT) and confocal microscopy to image roots grown in compacted soil conditions from a plant to a cellular scale. The method is suited to visualize cellular responses of root tips grown in both non-compacted and compacted soils. This protocol presents a fully integrated workflow, including soil column preparation, creation of compaction conditions, plant growth, imaging, and quantification of root adaptive responses at a cellular scale.
根系是植物吸收养分和水分的主要器官,因此对植物的生长发育至关重要。然而,异质环境的物理挑战和多样的土壤胁迫会影响土壤中根系的生长。土壤压实是一个严重的全球性问题,会抑制根的伸长,减少根表面积并影响根系觅食资源。由于土壤这种生长基质的不透明性,在土壤中对根系进行可视化和定量分析很困难;然而,对于致力于应对这一挑战的植物和土壤科学家来说,非破坏性成像技术现在越来越容易获得。我们最近开发了一种综合方法,将X射线计算机断层扫描(X射线CT)和共聚焦显微镜相结合,以对在压实土壤条件下从植株到细胞尺度生长的根系进行成像。该方法适用于可视化在非压实和压实土壤中生长的根尖的细胞反应。本方案展示了一个完全集成的工作流程,包括土柱制备、压实条件创建、植物生长、成像以及在细胞尺度上对根系适应性反应的定量分析。