Abstract
Background3D high-resolution X-ray imaging methods have emerged over the last years for visualising the anatomy of tissue samples without substantial sample preparation. Quantitative analysis of cells and intercellular spaces in these images has, however, been difficult and was largely based on manual image processing. We present here an automated procedure for processing high-resolution X-ray images of parenchyma tissues of apple (Malus × domestica Borkh.) and pear (Pyrus communis L.) as a rapid objective method for characterizing 3D plant tissue anatomy at the level of single cells and intercellular spaces.ResultsWe isolated neighboring cells in 3D images of apple and pear cortex tissues, and constructed a virtual sieve to discard incorrectly segmented cell particles or unseparated clumps of cells. Void networks were stripped down until their essential connectivity features remained. Statistical analysis of structural parameters showed significant differences between genotypes in the void and cell networks that relate to differences in aeration properties of the tissues.ConclusionsA new model for effective oxygen diffusivity of parenchyma tissue is proposed that not only accounts for the tortuosity of interconnected voids, but also for significant diffusion across cells where the void network is not connected. This will significantly aid interpretation and analysis of future tissue aeration studies. The automated image analysis methodology will also support pheno- and genotyping studies where the 3D tissue anatomy plays a role.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-015-0650-y) contains supplementary material, which is available to authorized users.
Highlights
3D high-resolution X-ray imaging methods have emerged over the last years for visualising the anatomy of tissue samples without substantial sample preparation
Characteristics of parenchyma cells in pome fruit cortex Images of fruit cortex parenchyma obtained at 5 μm pixel resolution can resolve characteristic features of the cell architecture and air spaces of apple and pear tissue (Fig. 1)
The methodology described in this article yields distributions of cell size and shape as well as a quantitative description of tissue architecture and the void network that can lead to better plant anatomy understanding and models
Summary
3D high-resolution X-ray imaging methods have emerged over the last years for visualising the anatomy of tissue samples without substantial sample preparation. We present here an automated procedure for processing high-resolution X-ray images of parenchyma tissues of apple (Malus × domestica Borkh.) and pear (Pyrus communis L.) as a rapid objective method for characterizing 3D plant tissue anatomy at the level of single cells and intercellular spaces. Bulky plant organs such as pome fruit mainly consist of parenchyma tissue, that is important for metabolic processes such as respiration. While tissue porosity can be measured, tortuosity depends on the structure of the pore network and is more difficult
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