Forty-two mutants of the anther smut fungus Ustilago violacea were mapped by means of complementation tests, mitotic haploidization, and meiotic segregation. Spontaneous mitotic haploidization was very rare, but haploids were induced at a high frequency using p–fluorophenylalanine (PFP). Haploid segregants appeared as fast-growing, spherical colonies (papillae) which grew away from the diploid growth on PFP medium. Thirty-three markers, classified by complementation tests into 21 genes, were mapped by mitotic haploidization in 10–12 linkage groups. There were no discrepancies in the linkage data, and all the markers could be assigned unequivocally to linkage groups. Although about 250 diploids were analysed, there were no segregants in which mitotic crossing-over and mitotic haploidization appeared to have occurred simultaneously.
Thirteen of the 33 markers, in six or seven genes, were expressed infrequently (0–5%) in the papillae produced on PFP medium. These markers, which behaved unusually and were designated missing-markers, were found to be on two chromosomes which tended to remain disomic on PFP medium. Thus 8–10 chromosomes haploidize readily on PFP medium, whereas two other chromosomes are resistant to the effects of PFP and remain disomic. Meiotic segregation was investigated in crosses of genetically marked haploid stocks and also hi diploids, using the host plant. Some of the results enabled preliminary maps to be made of three linkage groups. The results from meiotic segregation were fully compatible with those from mitotic haploidization and the complementation tests.
The genetical evidence for a haploid chromosome number of at least 10–12 is in conflict with the observations of several cytologists that n = 2 in this species.