The water and mineral conductive tube, the xylem vessel and tracheid, is a highly conspicuous tissue due to its elaborately patterned secondary-wall deposition. One constituent of the xylem vessel and tracheid, the tracheary element, is an empty dead cell that develops secondary walls in the elaborate patterns. The wall pattern is appropriately regulated according to the developmental stage of the plant. The cytoskeleton is an essential component of this regulation. In fact, the cortical microtubule is well known to participate in patterned secondary cell wall formation. The dynamic rearrangement of the microtubules and actin filaments have also been recognized in the cultured cells differentiating into tracheary elements in vitro. There has recently been considerable progress in our understanding of the dynamics and regulation of cortical microtubules, and several plant microtubule associated proteins have been identified and characterized. The microtubules have been observed during tracheary element differentiation in living Arabidopsis thaliana cells. Based on this recently acquired information on the plant cytoskeleton and tracheary element differentiation, this review discusses the role of the cytoskeleton in secondary cell wall formation.