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Updated: Jun 26, 2026
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A Robotic Platform for High-throughput Protoplast Isolation and Transformation
Published on: September 27, 2016
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Using A Protoplast Transformation System to Enable Functional Studies in Mangifera indica L
Mark Owusu Adjei1,2, Huan Zhao1, Xiaoguang Tao1
1College of Landscape Architecture, Sichuan Agricultural University, Chengdu 611130, China.
International Journal of Molecular Sciences
|August 12, 2023
Summary
Researchers optimized mango protoplast isolation and transformation for gene studies. This new method efficiently transforms mango protoplasts, aiding molecular mechanism research in this important fruit tree.
Area of Science:
- Plant Biotechnology
- Molecular Biology
- Horticulture
Background:
- Mango (Mangifera indica L.) transformation technology is underdeveloped.
- Protoplast technology offers a rapid method for transient expression and transformation assays, especially for plants lacking Agrobacterium-mediated systems.
Purpose of the Study:
- To optimize protoplast isolation and transformation conditions for mango.
- To establish an efficient system for mango gene expression regulation studies.
Main Methods:
- Optimized enzyme concentrations (cellulase R-10, macerozyme R-10) and digestion time for protoplast isolation.
- Determined optimal mannitol concentrations for protoplast yield, viability, and counting.
- Utilized polyethylene glycol (PEG)-4000-CaCl2 mediated transformation of isolated mango leaf protoplasts.
Main Results:
- Identified optimal conditions: 150 mg/mL cellulase R-10, 180 mg/mL macerozyme R-10, pH 5.6, 12h digestion.
- Achieved high protoplast yield with specific mannitol concentrations (0.16 M in wash, 0.08 M in MMG).
- Demonstrated 84.38% transformation efficiency using PEG-mediated transfection, with optimal mannitol for viability and successful GFP expression.
Conclusions:
- Developed a high-efficiency protocol for mango protoplast isolation and PEG-mediated transformation.
- This protocol provides an effective tool for rapid molecular mechanism studies in mango.
- The optimized system facilitates gene expression regulation research in mangoes.
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