Isolation of sterol mutants inChlamydomonas reinhardi: Chromatographic analyses

Abstract

Several sterol mutants ofChlamydomonas reinhardi were isolated by selecting for single colonies resistant to the polyene antibiotic, nystatin. The sterol profiles of three isolates-KD7, KD16, and KD21-as well as a wild type strain (137C) were determined by gas liquid chromatography and mass spectrometry. The wild type strain contained ergosterol (C28-5,7,22-triene) as the predominant sterol, with smaller amounts of a C28-5,7-diene, a C28-7-ene, and C-24 ethyl analogs of these three sterols. The three mutants had altered sterol composition, but none required exogenous sterol for growth. KD7 contained a C28-5,7,22,25-tetraene, a C28-5,7,25-triene, a C28-7,25-diene, and three C-24 ethyl analogs. The lesion in KD7 is apparently an inability to reduce the C-25 double bond required for the biosynthesis of ergosterol and an ethyl analog. The predominant sterols accumulated by KD16 and KD21 were a C28-5,7-diene, a C28-7-ene, and C-24 ethyl versions of these two sterols, suggesting that these strains are 22(23)-desaturase mutants.