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Updated: May 11, 2026
08:38
Scalable Transfection of Maize Mesophyll Protoplasts
Published on: June 23, 2023
[Optimized condition for protoplast isolation from maize, wheat and rice leaves].
1Biotechnology Research Institute, Chinese Academy ofAgricultural Sciences, Beijing 100081, China.
Summary
Optimizing enzyme concentration and isolation time significantly improves protoplast yield in maize, wheat, and rice. These refined methods enhance protoplast isolation for staple crop research.
Area of Science:
- Plant Biotechnology
- Molecular Biology
- Agricultural Science
Context:
- Maize, wheat, and rice are globally significant staple crops.
- Efficient protoplast isolation is crucial for genetic manipulation and crop improvement.
- Previous methods lacked optimized parameters for these specific crops.
Purpose:
- To determine the optimal conditions for maize, wheat, and rice protoplast isolation.
- To investigate the effects of enzyme concentration, isolation time, and centrifugal force on protoplast yield and viability.
- To establish reliable protocols for high-yield and high-viability protoplast isolation.
Summary:
- Orthogonal experimental design identified optimal enzyme concentrations (Cellulase R-10, Macerozyme R-10), incubation times, and centrifugal forces for each crop.
- Maize: 1.5% Cellulase R-10, 0.5% Macerozyme R-10, 7h, 100 x g; Wheat: 1.5% Cellulase R-10, 0.5% Macerozyme R-10, 5h, 100 x g; Rice: 2.0% Cellulase R-10, 0.7% Macerozyme R-10, 7h, 1000 x g.
- Protoplast yields ranged from 6-7x10^6 cells/g FW with over 90% viability, achieving 50-80% transformation efficiency using PEG-Ca2+.
Impact:
- Provides optimized, reproducible protocols for protoplast isolation in three major cereal crops.
- Facilitates advancements in genetic engineering, functional genomics, and breeding programs for maize, wheat, and rice.
- Enhances the efficiency of protoplast-based transformation systems for crop improvement research.
