TL;DR
Detailed instructions are provided for generating conditional gene deletions in the corn phytopathogenic fungus Ustilago maydis, while infecting corn plants through transcriptional control of the recombinase by fungal promoters that are specifically activated during the biotrophic phase of fungal growth.
This chapter provides a detailed, step-by-step protocol for implementing a conditional plasmid system that enables efficient, markerless gene deletion in FNA strains and provides a powerful and adaptable tool for advancing genetic studies in this genetically recalcitrant subspecies.
Plant transformation remains constrained by labor-intensive tissue culture. Our previous work showed that direct delivery of developmental regulators (DRs) can induce de novo meristems on plants, offering a promising transformation approach. In this resource article, we introduced DipTRANS (Direct in planta Transformat...
A novel marker recycling cassette is reported for Aspergillus species that enables highly efficient gene disruption and marker recycling by combining a doxycycline-inducible Cre/ loxP system with a uracil biosynthesis pathway and counterselection using uracil analogs.
A brief summary of the most important considerations for expression of heterologous genes, including expression strain selection, codon optimization, the choice of insertion loci, as well as potential protein fusions for targeting, localization, purification, and detection, are provided.
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