Parasexual crossings for bulk segregant analysis in Aspergillus niger to facilitate mutant identification via whole genome sequencing

Abstract

The industrially important fungus Aspergillus niger is known to reproduce only asexually. The parasexual cycle of fungi can be used for crossing two different strains to produce segregants or progeny with combined mutations even in fungi without a known sexual cycle. In A. niger, the parasexual cycle has been extensively used to establish linkage groups and to generate genetic maps. With the advent of whole genome sequencing, the parasexual cycle has received renewed attention as a method to create segregants for bulk segregant analysis. Bulk segregant analysis is a genetic technique used to link and ultimately identify the mutation associated with a particular phenotype. In this chapter we describe the procedure for setting up parasexual crossings in A. niger. The segregants obtained with this method can be used in combination with next-generation sequencing to map mutations in the organism.