Abstract
Direct visualization of the organization and dynamics of the actin cytoskeleton in rice cells is essential to understand its roles in regulating rice growth and development. Visualization of actin dynamics in protoplasts transformed with actin probe is a relatively quick and simple strategy, compared to the strategy of generating stable transgenic rice plants that harbor actin probe, which is time-consuming. Here is a protocol described in details regarding transforming rice protoplasts as well as visualizing and quantifying actin dynamics in rice protoplasts, which is based on the method previously reported (Shi et al., 2013).
Keywords: Rice protoplast, Actin dynamics, Filament severing frequency, Actin depolymerization rate, Filament elongation rate
Materials and Reagents
Equipment
Software
Procedure
Recipes
Acknowledgments
This protocol was adapted from our previously published work (Shi et al., 2013). We thank Meng Shi and Shaojie Cui for helpful suggestions on rice protoplast preparation and transformation. The research in the Huang lab was supported by grants from Ministry of Science of Technology (2013CB945100) and National Natural Science Foundation of China (31125004).
References
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