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Updated: Aug 2, 2026
A Robotic Platform for High-throughput Protoplast Isolation and Transformation
Published on: September 27, 2016
Efficient mesophyll protoplast isolation and development of a transient expression system for castor-oil plant
Ying Liu1, Yingbin Xue1, Jianian Tang1,2
1Department of Biotechnology, Faculty of Agricultural Science, Guangdong Ocean University, Zhanjiang, Guangdong, P. R. China.
Introduction:
We investigated the main factors affecting the efficacy of protoplast isolation, including leaf-obtaining period, cutting shapes of leaf material, enzyme concentration, enzymolysis time, and centrifugal speed.
Methods:
Protoplast isolation was optimal on the condition of 20 days of leaf materials, 2-mm filament of leaves, 1.6% RS and 0.8% R-10, 80 min of enzymolysis, and 700 rpm of centrifugation, resulting in the best yield (1.19 X 106 protoplasts/g FW) and vitality (80.34%) of mesophyll protoplasts. The transient expression vector pGFPl with green fluorescent protein was transfected into the obtained protoplasts from castor by polyethylene glycol-mediated method with a transformation efficiency of 12.37%.
Results:
Moreover, the applicability of the system for studying the subcellular localization of Re FATA (an acyl-ACP thioesterase) was validated via the protoplast isolation and transient expression protocol in this study.
Discussion:
Collectively, the efficient mesophyll protoplast isolation and protoplast transient expression system facilitate to analyze the function of specific gene in castor (Ricinus communis L).
