Nicotiana benthamiana as a model for studying Cryptococcus-plant interaction

Cryptococcus gattii, an environmental yeast isolated from plants, is one of the agents of cryptococcosis. Here, we aimed to develop a plant model to study C. gattii-plant interaction, since it is unclear how it affects the plant and the yeast. We tested three inoculation methods (scarification, infiltration, and abrasion) in three plant species: Arabidopsis thaliana, Nicotiana tabacum, and N. benthamiana. Cryptococcus gattii was able to grow in all three models, with a peak of yeast cell burden after 7 days, without any pathological effects. Furthermore, the fungal burden was reduced later, confirming that C. gattii is not a phytopathogen. Cryptococcus gattii proliferation was higher in N. benthamiana, which presented an increase in the hydrogen peroxide content, antioxidant system activity, and indoleacetic acid (IAA) production. Cryptococcus gattii colonies recovered from N. benthamiana presented lower ergosterol content, reduced capsule, and increased growth rate in vitro and inside macrophages. In vitro, IAA altered C. gattii morphology and susceptibility to antifungal drugs. We hypothesize that C. gattii can temporarily colonize plant living tissues, which can be a potential reservoir of yeast virulence, with further dissemination to the environment, birds, and mammals. In conclusion, N. benthamiana is suitable for studying C. gattii-plant interaction.