Diuron (3-[3,4-dichlorophenyl]-1,1-dimethylurea), an inhibitor of mitochondrial respiration, blocks the yeast respiratory chain between cytochrome b and c1. Diuron-resistant mutants of Saccharomyces cerevisiae have been selected and several mutations localized to the mitochondrial cytochrome b gene. The present paper identifies specific DNA base changes within the cytochrome b gene conferring diuron-resistance. DNA sequence analysis was done utilizing primer extension of crude mitochondrial RNA preparations in the presence of reverse transcriptase. Five independent diuron-resistant mutations have been sequenced.