Isolation and genetic analysis of mutant strains of Chlamydomonas reinhardi defective in gametic differentiation

Abstract

Impotent mutant strains of Chlamydomonas reinhardi, mating type (mt) plus, are described that have normal growth and motility but fail to differentiate into normal gametes. Procedures for their isolation and their genetic analysis are described. Five of the imp strains (imp 2, imp 5, imp 6, imp 7 and imp 8) exhibit no flagellar agglutination when mixed with mt- or mt+ gametes; these strains have been induced to form rare zygotes with mt- gametes and the mutations are shown to be unlinked to the mt locus (with the possible exception of imp 7). Two of the strains (imp 3 and imp 4) carry leaky mutations that affect cell fusion; neither mutation is found by tetrad analysis to be linked to mt or to the other. Cells of the imp 1 strain agglutinate well with mt- gametes and active agglutination continues for up to 48 hours, but cell fusion occurs only very rarely. Analysis of these rare zygotes indicates that imp 1 is closely linked to the mt+ locus, and fine structural studies reveal that imp 1 gametes produce a mutant mating structure involved in zygotic cell fusion. The development of sexuality in C. reinhardi therefore appears amenable to genetic dissection.