Improved media for testing the mating reaction and genetic complementation of Ustilago hordei

Abstract

The sexual cycle of Ustilago hordei, which results in the formation of teliospores, requires growth on its barley host for completion. However, the early steps of mating, including conjugation and the formation of dikaryotic mycelium, can occur on artificial media. The addition of activated charcoal to a variety of media enhanced the stability and intensity of the mating reaction as measured by mycelium formation. The incubation time at which the strongest mating reaction occurred was also reduced. The dikaryotic nature of the mycelia that resulted from mating on charcoal-containing media was confirmed by fluorescence microscopy. Complementation assays using minimal medium containing activated charcoal demonstrated allelism of mutations in auxotrophic sporidial strains of opposite mating type. The ease and reliability of this mating test allow for rapid identification of the mating type of unknown isolates and progeny of crosses, as well as providing a dependable procedure for performing complementation tests. Key words: barley, covered smut, Hordeum vulgare, mating type.