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Methods of In Situ Quantitative Root Biology
When dealing with plant roots, a multiscale description of the functional root structure is needed. Since the beginning of 21st century, new devices such as laser confocal microscopes have been accessible for coarse root structure measurements, including three-dimensional (3D) reconstruction. Most r...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8625443/ https://www.ncbi.nlm.nih.gov/pubmed/34834762 http://dx.doi.org/10.3390/plants10112399 |
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author | Pasternak, Taras Pérez-Pérez, José Manuel |
author_facet | Pasternak, Taras Pérez-Pérez, José Manuel |
author_sort | Pasternak, Taras |
collection | PubMed |
description | When dealing with plant roots, a multiscale description of the functional root structure is needed. Since the beginning of 21st century, new devices such as laser confocal microscopes have been accessible for coarse root structure measurements, including three-dimensional (3D) reconstruction. Most researchers are familiar with using simple 2D geometry visualization that does not allow quantitative determination of key morphological features from an organ-like perspective. We provide here a detailed description of the quantitative methods available for 3D analysis of root features at single-cell resolution, including root asymmetry, lateral root analysis, cell size and nuclear organization, cell-cycle kinetics, and chromatin structure analysis. Quantitative maps of the root apical meristem (RAM) are shown for different species, including Arabidopsis thaliana (L.), Heynh, Nicotiana tabacum L., Medicago sativa L., and Setaria italica (L.) P. Beauv. The 3D analysis of the RAM in these species showed divergence in chromatin organization and cell volume distribution that might be used to study root zonation for each root tissue. Detailed protocols and possible pitfalls in the usage of the marker lines are discussed. Therefore, researchers who need to improve their quantitative root biology portfolio can use them as a reference. |
format | Online Article Text |
id | pubmed-8625443 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-86254432021-11-27 Methods of In Situ Quantitative Root Biology Pasternak, Taras Pérez-Pérez, José Manuel Plants (Basel) Technical Note When dealing with plant roots, a multiscale description of the functional root structure is needed. Since the beginning of 21st century, new devices such as laser confocal microscopes have been accessible for coarse root structure measurements, including three-dimensional (3D) reconstruction. Most researchers are familiar with using simple 2D geometry visualization that does not allow quantitative determination of key morphological features from an organ-like perspective. We provide here a detailed description of the quantitative methods available for 3D analysis of root features at single-cell resolution, including root asymmetry, lateral root analysis, cell size and nuclear organization, cell-cycle kinetics, and chromatin structure analysis. Quantitative maps of the root apical meristem (RAM) are shown for different species, including Arabidopsis thaliana (L.), Heynh, Nicotiana tabacum L., Medicago sativa L., and Setaria italica (L.) P. Beauv. The 3D analysis of the RAM in these species showed divergence in chromatin organization and cell volume distribution that might be used to study root zonation for each root tissue. Detailed protocols and possible pitfalls in the usage of the marker lines are discussed. Therefore, researchers who need to improve their quantitative root biology portfolio can use them as a reference. MDPI 2021-11-06 /pmc/articles/PMC8625443/ /pubmed/34834762 http://dx.doi.org/10.3390/plants10112399 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Technical Note Pasternak, Taras Pérez-Pérez, José Manuel Methods of In Situ Quantitative Root Biology |
title | Methods of In Situ Quantitative Root Biology |
title_full | Methods of In Situ Quantitative Root Biology |
title_fullStr | Methods of In Situ Quantitative Root Biology |
title_full_unstemmed | Methods of In Situ Quantitative Root Biology |
title_short | Methods of In Situ Quantitative Root Biology |
title_sort | methods of in situ quantitative root biology |
topic | Technical Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8625443/ https://www.ncbi.nlm.nih.gov/pubmed/34834762 http://dx.doi.org/10.3390/plants10112399 |
work_keys_str_mv | AT pasternaktaras methodsofinsituquantitativerootbiology AT perezperezjosemanuel methodsofinsituquantitativerootbiology |