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Breaking of hyphae derived from growth of the phytopathogenic fungus Crinipellis perniciosa in liquid media yielded cell aggregates that performed as "quasi single cell" in toxicity assays. When treated with the chemical mutagens 4-nitroquinoleine-1-oxide (4NQO), hydrogen peroxide (H2O2), or paraquat (PAQ) as well as with ultraviolet light (UVC), broken hyphae of C. perniciosa gave a single cell-like response, i.e., survival curves similar to those obtained when treating single-cell suspensions. C. perniciosa had significantly higher UVC resistance than haploid bakers yeast but was more sensitive to 4NQO and extremely sensitive to PAQ and H2O2 when compared to likewise-treated yeast. Haploid C. perniciosa basidiospores (monokaryotic) were significantly more UVC resistant than C. perniciosa broken hyphae and than haploid and diploid yeast wild-type strains. This suggests a high capacity in C. perniciosa for repair of UVC-induced DNA lesions or, alternatively, an efficient protection from UVC irradiation, especially in basidiospores. The pronounced sensitivity of the dikaryotic form of C. perniciosa to PAQ and H2O2 points to a weak protection from oxidative stress-inducing agents.

Witches' broom disease (WBD) of cacao, caused by the hemibiotrophic fungus, Crinipellis perniciosa, exhibits a succession of symptoms that are caused by the biotrophic phase of the fungus. However, the study of this biotrophic phase is limited by its exclusive growth inside the plant or in the presence of callus. Here we report for the first time a method for the growth and maintenance of the biotrophic-like phase of C. perniciosa on a defined medium with metabolites found in the diseased tissues. Our results suggest that glycerol is a key carbon source for this interaction. This is a crucial achievement toward understanding the biology of this fungus during the infectious phase of WBD.