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Updated: Aug 8, 2026
A Simple Method for Isolation of Soybean Protoplasts and Application to Transient Gene Expression Analyses
Published on: January 25, 2018
Isolation of mesophyll protoplasts from mature leaves of soybeans
1Central Research and Development Department, Experimental Station, E. I. du Pont de Nemours and Company, Wilmington, Delaware 19898.
Abstract:
A procedure based on a combined cellulase-Pectolyase Y-23 enzyme digestion and metrizamide-sorbitol gradient purification protocol was developed for isolating mesophyll protoplasts from mature leaves of soybean (Glycine max L. Merr.). Based on chlorophyll content, this procedure results in a 10 to 15% protoplast yield from fully expanded mature leaves and a 20 to 30% yield from young (expanding) leaves within 3 hours. Isolated protoplasts displayed high rates of HCO(3) (-)-dependent photosynthesis; greater than 75 micromoles O(2) evolved per milligram chlorophyll per hour at 25 degrees C. This photosynthetic rate is comparable to that of mesophyll cells isolated mechanically from the same leaves.
