Isolation of Mutants and Construction of Intersexual Heterokaryons of Blakeslea trispora

The Mucoral fungus Blakeslea trispora is used for the industrial production of β-carotene and lycopene. Two genetic techniques have been used to increase carotene accumulation: the isolation of mutants and the formation and segregation of heterokaryons. Because all life stages are multinucleated, recessive mutants are isolated after exposure to N -methyl-N ′-nitro-N -nitrosoguanidine, a strong mutagen and inactivator of nuclei. Intersexual heterokaryons are obtained easily, because they are formed spontaneously during sexual interaction. Here are the pertaining methods, based on those previously developed for Phycomyces blakesleeanus , a related and better-known fungus.