Alfalfa Root Growth Rate Correlates with Progression of Microtubules during Mitosis and Cytokinesis as Revealed by Environmental Light-Sheet Microscopy

Cell division and expansion are two fundamental biological processes supporting indeterminate root growth and development of plants. Quantitative evaluations of cell divisions related to root growth analyses have been performed in several model crop and non-crop plant species, but not in important l...

Full description

Bibliographic Details
Main Authors: Petra Vyplelová, Miroslav Ovečka, Jozef Šamaj
Format: Article
Language:English
Published: Frontiers Media S.A. 2017-10-01
Series:Frontiers in Plant Science
Subjects:
Online Access:http://journal.frontiersin.org/article/10.3389/fpls.2017.01870/full
_version_ 1818652743240253440
author Petra Vyplelová
Miroslav Ovečka
Jozef Šamaj
author_facet Petra Vyplelová
Miroslav Ovečka
Jozef Šamaj
author_sort Petra Vyplelová
collection DOAJ
description Cell division and expansion are two fundamental biological processes supporting indeterminate root growth and development of plants. Quantitative evaluations of cell divisions related to root growth analyses have been performed in several model crop and non-crop plant species, but not in important legume plant Medicago sativa. Light-sheet fluorescence microscopy (LSFM) is an advanced imaging technique widely used in animal developmental biology, providing efficient fast optical sectioning under physiological conditions with considerably reduced phototoxicity and photobleaching. Long-term 4D imaging of living plants offers advantages for developmental cell biology not available in other microscopy approaches. Recently, LSFM was implemented in plant developmental biology studies, however, it is largely restricted to the model plant Arabidopsis thaliana. Cellular and subcellular events in crop species and robust plant samples have not been studied by this method yet. Therefore we performed LSFM long-term live imaging of growing root tips of transgenic alfalfa plants expressing the fluorescent molecular marker for the microtubule-binding domain (GFP-MBD), in order to study dynamic patterns of microtubule arrays during mitotic cell division. Quantitative evaluations of cell division progress in the two root tissues (epidermis and cortex) clearly indicate that root growth rate is correlated with duration of cell division in alfalfa roots. Our results favor non-invasive environmental LSFM as one of the most suitable methods for qualitative and quantitative cellular and developmental imaging of living transgenic legume crops.
first_indexed 2024-12-17T02:26:51Z
format Article
id doaj.art-d9b9ea5270824f0183a4a0876a49ddac
institution Directory Open Access Journal
issn 1664-462X
language English
last_indexed 2024-12-17T02:26:51Z
publishDate 2017-10-01
publisher Frontiers Media S.A.
record_format Article
series Frontiers in Plant Science
spelling doaj.art-d9b9ea5270824f0183a4a0876a49ddac2022-12-21T22:07:04ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2017-10-01810.3389/fpls.2017.01870304804Alfalfa Root Growth Rate Correlates with Progression of Microtubules during Mitosis and Cytokinesis as Revealed by Environmental Light-Sheet MicroscopyPetra VyplelováMiroslav OvečkaJozef ŠamajCell division and expansion are two fundamental biological processes supporting indeterminate root growth and development of plants. Quantitative evaluations of cell divisions related to root growth analyses have been performed in several model crop and non-crop plant species, but not in important legume plant Medicago sativa. Light-sheet fluorescence microscopy (LSFM) is an advanced imaging technique widely used in animal developmental biology, providing efficient fast optical sectioning under physiological conditions with considerably reduced phototoxicity and photobleaching. Long-term 4D imaging of living plants offers advantages for developmental cell biology not available in other microscopy approaches. Recently, LSFM was implemented in plant developmental biology studies, however, it is largely restricted to the model plant Arabidopsis thaliana. Cellular and subcellular events in crop species and robust plant samples have not been studied by this method yet. Therefore we performed LSFM long-term live imaging of growing root tips of transgenic alfalfa plants expressing the fluorescent molecular marker for the microtubule-binding domain (GFP-MBD), in order to study dynamic patterns of microtubule arrays during mitotic cell division. Quantitative evaluations of cell division progress in the two root tissues (epidermis and cortex) clearly indicate that root growth rate is correlated with duration of cell division in alfalfa roots. Our results favor non-invasive environmental LSFM as one of the most suitable methods for qualitative and quantitative cellular and developmental imaging of living transgenic legume crops.http://journal.frontiersin.org/article/10.3389/fpls.2017.01870/fullcell divisiondevelopmental imaginglight-sheet microscopyMedicago sativamicrotubulesroot growth
spellingShingle Petra Vyplelová
Miroslav Ovečka
Jozef Šamaj
Alfalfa Root Growth Rate Correlates with Progression of Microtubules during Mitosis and Cytokinesis as Revealed by Environmental Light-Sheet Microscopy
Frontiers in Plant Science
cell division
developmental imaging
light-sheet microscopy
Medicago sativa
microtubules
root growth
title Alfalfa Root Growth Rate Correlates with Progression of Microtubules during Mitosis and Cytokinesis as Revealed by Environmental Light-Sheet Microscopy
title_full Alfalfa Root Growth Rate Correlates with Progression of Microtubules during Mitosis and Cytokinesis as Revealed by Environmental Light-Sheet Microscopy
title_fullStr Alfalfa Root Growth Rate Correlates with Progression of Microtubules during Mitosis and Cytokinesis as Revealed by Environmental Light-Sheet Microscopy
title_full_unstemmed Alfalfa Root Growth Rate Correlates with Progression of Microtubules during Mitosis and Cytokinesis as Revealed by Environmental Light-Sheet Microscopy
title_short Alfalfa Root Growth Rate Correlates with Progression of Microtubules during Mitosis and Cytokinesis as Revealed by Environmental Light-Sheet Microscopy
title_sort alfalfa root growth rate correlates with progression of microtubules during mitosis and cytokinesis as revealed by environmental light sheet microscopy
topic cell division
developmental imaging
light-sheet microscopy
Medicago sativa
microtubules
root growth
url http://journal.frontiersin.org/article/10.3389/fpls.2017.01870/full
work_keys_str_mv AT petravyplelova alfalfarootgrowthratecorrelateswithprogressionofmicrotubulesduringmitosisandcytokinesisasrevealedbyenvironmentallightsheetmicroscopy
AT miroslavovecka alfalfarootgrowthratecorrelateswithprogressionofmicrotubulesduringmitosisandcytokinesisasrevealedbyenvironmentallightsheetmicroscopy
AT jozefsamaj alfalfarootgrowthratecorrelateswithprogressionofmicrotubulesduringmitosisandcytokinesisasrevealedbyenvironmentallightsheetmicroscopy