We previously found that sucrose solution immersion treatment permitted ectopic guard cell differentiation, resulting in clustered stomatal guard cells. Using this system, we examined the effects of sucrose solution-induced stomatal clustering on guard cell cortical microtubules and the stomatal response to fusicoccin. Confocal observation revealed that the radial orientation of cortical microtubules was largely maintained in clustered guard cells. Outward movement of cortical microtubule plus-ends was also kept in the clustered guard cells. Fusicoccin treatment induced stomatal opening in both spaced and clustered stomata, although sucrose solution-treated guard cells had lower stomatal apertures. These results suggested that immersion treatment with sucrose solution perturbed the one-cell spacing of stomata but not the cortical microtubule organization required to open stomatal pores.
Keywords: Arabidopsis; fusicoccin; guard cells; image analysis; live imaging; microtubules; stomata.