Live Cell Imaging of Microtubule Cytoskeleton and Micromechanical Manipulation of the Arabidopsis Shoot Apical Meristem

This protocol it helps us understand how Microtubule Cytoskeleton behavior occurs in plants. At the same time, it also helps us understand how these structural components that influence cell and organ shape, respond to changes in mechanical forces. Although, this method requires little sophistication, it provides robust, quantitative read-after, the mechanical response differences between different genotypes and conditions.

Begin by growing arabidopsis seeds expressing microtubule binding domains fused with green fluorescent protein in soil, at 20 and 6 degrees Celsius under long day conditions for one week. After germination, transfer the seedlings into new pots with sufficient growth space to allow robust vegetative growth, and place the plants at 20 and 16 degrees Celsius under short day conditions. After three to five weeks, transfer the plants back to long day conditions at the same temperature until the plants bolt, allowing the inflorescence to grow up to two to five centimeters long.