Mitotic mapping in linkage group V of Aspergillus niger based on selection of auxotrophic recombinants by Novozym enrichment

Abstract

This paper describes a procedure which allows the quantitative selection of auxotrophs of the fungus Aspergillus niger by enzymatic killing of immobilized germinating prototrophic conidiospores. We have applied this procedure to linkage analysis on the basis of mitotic crossing-over in this fungus. Starting with a heterozygous diploid strain, we could select auxotrophic homozygous diploid recombinants quantitatively. We estimated the frequency of crossing-over after correction for clonal distribution of recombinants, and localized four auxotrophic markers as well as the centromere on chromosome V of this fungus. The Novozym enrichment procedure proved to be useful in genetic analysis and for the construction of recombinant genotypes in the case of closely linked auxotrophic markers. The detemination of gene order and the estimation of distances on the basis of benomyl-induced recombinant haploid segregants may lead to incorrect conclusions. Genetic analysis on the basis of homozygous recombinants, however, can provide reliable estimates of map distances.Key words: Aspergillus niger, chromosome mapping, mitotic crossing-over, Novozym enrichment, auxotrophic recombinants.