Alternative oxidase (AOX) constitutes a nonenergy conserving branch of the mitochondrial electron transport chain. AOX activity may be important to avoid reactive oxygen species (ROS) generation by the chain, particularly during abiotic stress. We compared leaf AOX1a transcript and AOX protein amounts in wild-type (WT) Nicotiana tabacum plants experiencing mild to severe drought. Mild to moderate drought resulted in a progressive and modest increase in AOX amount, accompanied by a progressive increased expression of different ROS-scavenging components. Under these conditions, transgenic plants with suppressed AOX amount, due to an RNA interference construct, were not compromised in their ability to manage ROS load and prevent cellular damage. Severe drought stress, particularly when combined with increased irradiance, strongly increased AOX amount in WT tobacco and this coincided with an increase in total respiration and, despite further induction of ROS-scavenging systems, some evidence of cellular damage. Under these severe conditions, plants lacking AOX suffered more cellular damage than WT and, at the most severe stage, were found to downregulate rather than upregulate the transcript level of several important ROS-scavenging components. At this stage, WT plants could still recover rapidly after rewatering, but the recoverability of AOX knockdown plants was strongly compromised.