A delta-9 fatty acid desaturase gene, homologous to animal and fungal acyl-coenzyme A (CoA) desaturases, was isolated from the red alga Cyanidioschyzon merolae using a degenerate PCR strategy. This gene, designated as CmFAD9, has no intron. The encoded delta-9 fatty acid desaturase (CmFad9p) consists of 476 amino acids and has an estimated molecular mass of 55.4 kDa. CmFad9p is a unique delta-9 fatty acid desaturase among plants, in that it is fused with the cytochrome b5 domain at its carboxyl terminus. This is characteristic of yeast acyl-CoA desaturase. Genomic Southern hybridization suggested that the C. merolae genome contains a single gene for delta-9 fatty acid desaturase of the animal and fungal type. Southern hybridization combined with pulsed-field gel electrophoresis revealed that CmFAD9 is probably located on chromosome XI of the 17 C. merolae chromosomes. A 1.6-kb product of this gene was transcribed throughout a light/dark synchronization culture. The discovery of CmFAD9 indicates the existence of a novel type of plant delta-9 fatty acid desaturase that may function in the endoplasmic reticulum, but not in the plastid.