A filamentous fungus, Mortierella alpina 1S-4, was obtained, through extensive screening, as an potential producer of triacylglycerol containing C20 polyunsaturated fatty acids (PUFAs) such as arachidonic acid. With this discovery as a starting point, we conducted employing methods from metabolic engineering and molecular biology for controlling cultures and breeding mutant strains. These parental and mutant strains are now used for large-scale production of a variety of PUFAs.