Arabidopsis Leaf Protoplasts
Arabidopsis Leaf Protoplasts - Related Videos
07:42
Rapid Analysis of Circadian Phenotypes in Arabidopsis Protoplasts Transfected with a Luminescent Clock Reporter
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Cited by 18 •
2016
The circadian clock regulates about a third of the Arabidopsis transcriptome, but the percentage of genes that feed back into timekeeping remains unknown. Here we visualize a method to rapidly assess circadian phenotypes in any mutant line of Arabidopsis using luminescent imaging of a circadian reporter transiently expressed in protoplasts.
11:16
Isolation of Protoplasts from Tissues of 14-day-old Seedlings of Arabidopsis thaliana
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Cited by 59 •
2009
This video shows a procedure for isolating intact protoplasts from tissues of 14-day-old seedlings of Arabidopsis. Given that the isolated protoplasts remain intact for at least 96h and are isolated from seedlings instead of one-month-old mature plants, this procedure expedites assays requiring intact protoplasts.
02:58
An Assay to Detect Oxidative Burst in Arabidopsis Leaf Discs upon Immune Elicitation
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2025
This video demonstrates an assay to measure oxidative burst in Arabidopsis leaves upon immune elicitation. Leaf discs, cut from plant leaves, are incubated with bacterial peptides functioning as microbe-associated molecular patterns or MAMPs. Upon detection of the MAMPs, plant cells induce the production of extracellular ROS, which is detected using a chemiluminescence assay.
12:29
mRNA Interactome Capture from Plant Protoplasts
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Cited by 1 •
2017
Here, we present an interactome capture protocol applied to Arabidopsis thaliana leaf mesophyll protoplasts. This method critically relies on in vivo UV crosslinking and allows for the isolation and identification of plant mRNA-binding proteins from a physiological environment.
09:56
A Stepwise Guide to the Isolation and Analysis of Leaf Surface and Apoplastic RNA Using Arabidopsis Rosettes
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2025
We describe a simple yet highly reproducible method to isolate extracellular RNA from the leaf surface and apoplast of Arabidopsis plants.
FAQs
The osmoticum helps balance the conditions created when cellulase and pectinase remove the cell wall. Without that support, the exposed plasma membrane would be more vulnerable during isolation and handling. This pairing is therefore central to obtaining intact cells rather than merely digesting leaf tissue. Preserving membrane integrity improves the reliability of downstream manipulation and culture.
Cellulase and pectinase contribute to wall digestion, but their shared purpose is not simply tissue breakdown. Removing the wall exposes the plasma membrane, making the resulting cells accessible to introduced molecules and easier to manipulate in culture. This change in physical accessibility helps explain why protoplasts serve as a practical platform for rapid molecular experiments.
Transient gene expression, genome editing, and protein localization address different experimental questions. Transient expression supports rapid testing of introduced genetic information, protein localization reveals where a protein is found within the cell, and genome editing enables functional testing through genetic changes. Using the same cell system for these approaches helps connect molecular observations with gene or protein function.
A basic workflow starts with Arabidopsis leaves, uses cellulase and pectinase to digest cell walls, and includes an osmoticum to help preserve membrane integrity. The isolated cells can then be maintained in culture and manipulated for molecular assays, including introduction of molecules. This workflow produces a relatively uniform cell population suitable for rapid, comparable experiments.
Researchers would choose this system when they need rapid functional information from many comparable plant cells rather than relying only on whole-leaf observations. It supports transient gene expression, protein localization, and genome editing, allowing investigators to test genes, examine protein distribution, or assess regulatory pathways. The resulting experiments can provide focused evidence about cellular mechanisms.
In plant biology, these protoplasts allow cellular measurements to be linked with broader physiological questions. Researchers can analyze signaling, metabolism, and stress responses in a controlled cell population, then use the results to investigate how molecular mechanisms contribute to plant physiology. Their value lies in combining experimental flexibility with a direct connection between cellular behavior and biological function.