High-Resolution Whole-Mount Imaging of Three-Dimensional Tissue Organization and Gene Expression Enables the Study of Phloem Development and Structure inArabidopsis
- 1 June 2008
- journal article
- Published by Oxford University Press (OUP) in Plant Cell
- Vol. 20 (6), 1494-1503
- https://doi.org/10.1105/tpc.107.056069
Abstract
Currently, examination of the cellular structure of plant organs and the gene expression therein largely relies on the production of tissue sections. Here, we present a staining technique that can be used to image entire plant organs using confocal laser scanning microscopy. This technique produces high-resolution images that allow three-dimensional reconstruction of the cellular organization of plant organs. Importantly, three-dimensional domains of gene expression can be analyzed with single-cell precision. We used this technique for a detailed examination of phloem cells in the wild type and mutants. We were also able to recognize phloem sieve elements and their differentiation state in any tissue type and visualize the structure of sieve plates. We show that in the altered phloem development mutant, a hybrid cell type with phloem and xylem characteristics develops from initially normally differentiated protophloem cells. The simplicity of sieve element data collection allows for the statistical analysis of structural parameters of sieve plates, essential for the calculation of phloem conductivity. Taken together, this technique significantly improves the speed and accuracy of the investigation of plant growth and development.Keywords
This publication has 26 references indexed in Scilit:
- A Plasma Membrane-Anchored Fluorescent Protein Fusion Illuminates Sieve Element Plasma Membranes in Arabidopsis and TobaccoPlant Physiology, 2008
- Visualizing Plant Development and Gene Expression in Three Dimensions Using Optical Projection TomographyPlant Cell, 2006
- GAL4-GFP enhancer trap lines for genetic manipulation of lateral root development in Arabidopsis thalianaJournal of Experimental Botany, 2005
- Marking cell lineages in living tissuesThe Plant Journal, 2005
- OsiriX: An Open-Source Software for Navigating in Multidimensional DICOM ImagesJournal of Digital Imaging, 2004
- High-Throughput Fluorescent Tagging of Full-Length Arabidopsis Gene Products in PlantaPlant Physiology, 2004
- Application of a Single-solute Non-steady-state Phloem Model to the Study of Long-distance Assimilate TransportJournal of Theoretical Biology, 2003
- An aniline blue staining procedure for confocal microscopy and 3D imaging of normal and perturbed cellular phenotypes in mature Arabidopsis embryosThe Plant Journal, 2000
- Pattern of Differentiation of the First Vascular Elements in the Embryo and Seedling ofArabidopsis thalianaInternational Journal of Plant Sciences, 1999
- Phloem Loading by the PmSUC2 Sucrose Carrier from Plantago major Occurs into Companion CellsPlant Cell, 1995