Temperature stress such as heat, cold, or freezing is a principal cause for yield reduction in crops. In particular, heat stress is very common and dangerous for plants since this stress can impact several plant and cellular functions. In spite of their role in sensing local stress and in controlling fundamental processes including PCD, the responses of cellular structures and organelles to heat stress are poorly investigated. In this work, we investigated the possible changes induced by mild heat stress, medium heat stress, and heat shock (HS; 5 min at 35 degrees C, 45 degrees C, or 50 degrees C, respectively) on actin cytoskeleton and endoplasmic reticulum (ER) of tobacco BY-2 cultured cells. While mild and medium heat stresses are ineffective, HS induces depolymerization of actin microfilaments and changes in ER morphology accompanied by accumulation of the HSP70 binding protein (BiP). These effects of HS are prevented by the inhibitor of ethylene production Co(2+). While the analyzed cell structures do not seem to be involved in the establishment of mild and medium heat stresses at least in this experimental system, the strong modifications induced by the treatment at 50 degrees C suggest that actin cytoskeleton and ER may be involved in the responses to HS. Besides, the inhibiting effect of Co(2+) suggests a role for ethylene as a regulative molecule in the responses to HS here observed.