Back to Search
Start Over
High-resolution confocal imaging of wall ingrowth deposition in plant transfer cells: Semi-quantitative analysis of phloem parenchyma transfer cell development in leaf minor veins of Arabidopsis
- Source :
- BMC Plant Biology
- Publication Year :
- 2015
- Publisher :
- BioMed Central, 2015.
-
Abstract
- Background Transfer cells (TCs) are trans-differentiated versions of existing cell types designed to facilitate enhanced membrane transport of nutrients at symplasmic/apoplasmic interfaces. This transport capacity is conferred by intricate wall ingrowths deposited secondarily on the inner face of the primary cell wall, hence promoting the potential trans-membrane flux of solutes and consequently assigning TCs as having key roles in plant growth and productivity. However, TCs are typically positioned deep within tissues and have been studied mostly by electron microscopy. Recent advances in fluorophore labelling of plant cell walls using a modified pseudo-Schiff-propidium iodide (mPS-PI) staining procedure in combination with high-resolution confocal microscopy have allowed visualization of cellular details of individual tissue layers in whole mounts, hence enabling study of tissue and cellular architecture without the need for tissue sectioning. Here we apply a simplified version of the mPS-PI procedure for confocal imaging of cellulose-enriched wall ingrowths in vascular TCs at the whole tissue level. Results The simplified mPS-PI staining procedure produced high-resolution three-dimensional images of individual cell types in vascular bundles and, importantly, wall ingrowths in phloem parenchyma (PP) TCs in minor veins of Arabidopsis leaves and companion cell TCs in pea. More efficient staining of tissues was obtained by replacing complex clearing procedures with a simple post-fixation bleaching step. We used this modified procedure to survey the presence of PP TCs in other tissues of Arabidopsis including cotyledons, cauline leaves and sepals. This high-resolution imaging enabled us to classify different stages of wall ingrowth development in Arabidopsis leaves, hence enabling semi-quantitative assessment of the extent of wall ingrowth deposition in PP TCs at the whole leaf level. Finally, we conducted a defoliation experiment as an example of using this approach to statistically analyze responses of PP TC development to leaf ablation. Conclusions Use of a modified mPS-PI staining technique resulted in high-resolution confocal imaging of polarized wall ingrowth deposition in TCs. This technique can be used in place of conventional electron microscopy and opens new possibilities to study mechanisms determining polarized deposition of wall ingrowths and use reverse genetics to identify regulatory genes controlling TC trans-differentiation. Electronic supplementary material The online version of this article (doi:10.1186/s12870-015-0483-8) contains supplementary material, which is available to authorized users.
- Subjects :
- Propidium iodide
Confocal
Arabidopsis
Plant Science
law.invention
Cell wall
Confocal microscopy
law
Cell Wall
Parenchyma
Schiff Bases
Microscopy, Confocal
biology
Phloem parenchyma
Methodology Article
fungi
Transfer cells
Transfer cell
Confocal imaging
Anatomy
Pseudo-Schiff base
Companion cells
biology.organism_classification
Vascular bundle
Plant Leaves
Cell wall ingrowths
Biophysics
Phloem
Propidium
Subjects
Details
- Language :
- English
- ISSN :
- 14712229
- Volume :
- 15
- Database :
- OpenAIRE
- Journal :
- BMC Plant Biology
- Accession number :
- edsair.doi.dedup.....98915325421bf28baacbdaa6fa744240