Identification of Aspergillus nidulans genes essential for the accumulation of sterigmatocystin

The fungus Aspergillus nidulans (Emericella nidulans) was used as a genetic model for the identification of genes required for efficient accumulation of sterigmatocystin (ST). The required gene for sterigmatocystin expression was stc, which is an intermediate penultimate product in the aflatoxin biosynthetic pathway. Genetic analysis included studies of the sexual and parasexual cycles. The allelic segregation rates and recombination frequencies between linked and nonlinked genetic markers were determined by the crossing of the strains UT448 (stc) to UT196 (stc(+)) and UT448 (stc) to UT184 (stc). Low ST accumulation (4.0 ppm) in the UT196 strain and in 7.4% of the meiotic segregants allowed us to map the stc locus at chromosome I, 3.4% distant from riboA1. The diploid UT448 (stc)//UT184 (stc), prepared from nonproducing strains, was analyzed based on the parasexual cycle, and 28% of the haploid segregants accumulated the ST toxin. The results suggest that UT448 carries the stc mutant (or an inactivated) allele and that UT184, although carrying the stc(+) allele, is reactivated only by the R2(+) factor, which is located at chromosome VIII of UT448. In such a configuration, the diploid accumulates large amounts of sterigmatocystin (40 ppm). Another regulator factor (R1), located at the meth-w (II) chromosomic interval, was identified in the UT448 strain. At DNA level in chromosome I, the R1 product acts and blocks the stcZ(+) gene transcription. In a different genotypic configuration, the R1 product interacts with the R2 product (of chromosome VIII), allowing the stcZ(+) gene expression. Furthermore, the diploid UT448 (stc)//UT196 (stc(+)) accumulated the ST toxin at high level (40 ppm), indicating similar interaction between mentioned factors and the stc gene. Obtained data suggest that R1 (II) regulates the stcZ(+) transcription, by interacting with chromosome I (at the DNA level) and that R2 (VIII) controls R1 activity at the cytoplasm level. Based on these results, we propose a regulation model for the sterigmatocystin production.