Abstract
The need exists to create the auxotrophic mutants for basidiomycete Ustilago maydis to allow selection of gene transformants in minimal growth media. The ADE2 gene was identified by homology with Saccharomyces cerevisiae. Adenine-requiring auxotrophic mutants were effectively created by homologous recombination in protoplasts using a standard plasmid containing the ADE2 locus interrupted by a phleomycin resistance gene, selected on standard complex media. The knockout ade2 mutant produced grows on minimal, defined medium only if supplemented with adenine.
Highlights
Phytopathogenic Ustilago maydis is a haploid, dimorphic fungal model organism [1] that has been used to characterize genes involved in its infection of maize [2], in DNA recombination, mating determination and various signaling pathways [3,4]
Adenine auxotrophic mutants have been developed by knocking out a putative ADE6 in Ustilago strain 521 [13]
We identified homologous genes for the yeast histidine HIS3 loci YOR202w and YNL338w but were unable to obtain knockout transformants that were dependent on histidine in minimal media, possibly due to differences in metabolic pathways between Ustilago and yeast
Summary
Phytopathogenic Ustilago maydis is a haploid, dimorphic fungal model organism [1] that has been used to characterize genes involved in its infection of maize [2], in DNA recombination, mating determination and various signaling pathways [3,4]. In need for new selection alternatives to create additional gene transformants on minimal media in strains that already contained the carboxin-resistance insert, we turned to the auxotrophic mutant option. Few auxotrophic mutants have been described for Ustilago, likely because they cannot be used for selection in planta.
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