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Updated: Apr 1, 2026
11:16
Isolation of Protoplasts from Tissues of 14-day-old Seedlings of Arabidopsis thaliana
Published on: August 17, 2009
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Isolation of Chloroplasts from Plant Protoplasts
Shiu-Cheung Lung1, Matthew D Smith2, Simon D X Chuong3
1School of Biological Sciences, University of Hong Kong, Pokfulam, Hong Kong SAR, China;
Cold Spring Harbor Protocols
|October 3, 2015
Summary
This study presents an optimized protocol for isolating high-yield, pure chloroplasts from Arabidopsis protoplasts. The method ensures intact chloroplasts suitable for various scientific analyses.
Area of Science:
- Plant Biology
- Cell Biology
- Biochemistry
Background:
- Direct isolation of chloroplasts from plants often yields low purity and intactness.
- Existing methods require substantial starting material.
Purpose of the Study:
- To develop an optimized protocol for high-yield isolation of pure, intact chloroplasts.
- To provide a simple, scaled-down, and low-cost method for chloroplast isolation.
Main Methods:
- Isolation of healthy mesophyll protoplasts from Arabidopsis.
- A two-part method involving protoplast rupture to release intact chloroplasts.
Main Results:
- Achieved high yield and purity of isolated chloroplasts.
- Isolated chloroplasts demonstrated intactness and suitability for analyses.
Conclusions:
- The optimized protocol effectively yields pure, intact chloroplasts from Arabidopsis protoplasts.
- This method is suitable for biochemical, cellular, and molecular analyses.
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