Isolation and culture of monokaryotic haplonts of Ustilago nuda, the role of proline in their metabolism, and the inoculation of barley with resynthesized dikaryons

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Abstract
Spores of Ustilago nuda were germinated on nutrient medium containing benzoic acid or derivatives. These compounds inhibited mitotic divisions of the nuclei in the two dikaryons formed by one spore. After removal of the chemical from the medium, mitosis occurred and the four monokaryotic haplonts were isolated by microsurgery. The haplonts were cultured on media supplemented with L-proline, because all haplonts of (+) mating type, originating from spores collected in Europe, Asia, Africa, and North and South America, were found to be proline deficient. The gene controlling proline synthesis behaved as though it was identical with or part of the mating locus. Barley flowers at mid-anthesis were inoculated successfully with dikaryotic mycelium resynthesized from isolated haplonts.

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