Transcriptional Activator UvXlnR Is Required for Conidiation and Pathogenicity of Rice False Smut Fungus Ustilaginoidea virens.

Transcription factors play critical roles in diverse biological processes in fungi. XlnR, identified as a transcriptional activator that regulates the expression of the extracellular xylanase genes in fungi, has not been extensively studied for its function in fungal development and pathogenicity in rice false smut fungus Ustilaginoidea virens. In this study, we characterized UvXlnR in U. virens and established that the full-length, N-terminal, and C-terminal forms have the ability to activate transcription. The study further demonstrated that UvXlnR plays crucial roles in various aspects of U. virens biology. Deletion of UvXlnR affected growth, conidiation, and stress response. UvXlnR mutants also exhibited reduced pathogenicity, which could be partially attributed to the reduced expression of xylanolytic genes and extracellular xylanase activity of U. virens during the infection process. Our results indicate that UvXlnR is involved in regulating growth, conidiation, stress response, and pathogenicity.

The Bax inhibitor UvBI-1, a negative regulator of mycelial growth and conidiation, mediates stress response and is critical for pathogenicity of the rice false smut fungus Ustilaginoidea virens.

Bax inhibitor-1 (BI-1), an evolutionarily conserved protein, is a suppressor of cell death induced by the proapoptotic protein Bax and is involved in the response to biotic and abiotic stress in animals, plants and yeast. Rice false smut caused by Ustilaginoidea virens is one of the destructive rice diseases worldwide. Although BI-1 proteins are widely distributed across filamentous fungi, few of them are functionally characterized. In this study, we identified a BI-1 protein in U. virens, UvBI-1, which contains a predicted Bax inhibitor-1-like family domain and could suppress the cell death induced by Bax. By co-transformation of the CRISPR/Cas9 construct along with donor DNA fragment containing the hygromycin resistance gene, we successfully generated Uvbi-1 deletion mutants. The UvBI-1 deletion showed an increase in mycelia vegetative growth and conidiation, suggesting this gene acts as a negative regulator of the growth and conidiation. In addition, the Uvbi-1 mutants exhibited higher sensitivity to osmotic and salt stress, hydrogen peroxide stress, and cell wall or membrane stress than the wild-type strain. Furthermore, UvBI-1 deletion was found to cause increased production of secondary metabolites and loss of pathogenicity of U. virens. Taken together, our results demonstrate that UvBI-1 plays a negative role in mycelial growth and conidiation, and is critical for stress tolerance, cell wall integrity, secondary metabolites production and pathogenicity of U. virens. Therefore, this study provides new evidence on the conserved function of BI-1 among fungal organisms and other species.

Correction: Development of rice conidiation media for Ustilaginoidea virens.

[This corrects the article DOI: 10.1371/journal.pone.0217667.].

Development of rice conidiation media for Ustilaginoidea virens.

Rice false smut, caused by the ascomycete Ustilaginoidea virens, is a serious disease of rice worldwide. Conidia are very important infectious propagules of U. virens, but the ability of pathogenic isolates to produce conidia frequently decreases in culture, which influences pathogenicity testing. Here, we developed tissue media with rice leaves or panicles that stimulate conidiation of U. virens. Among the tested media, 0.10 g/ml panicle medium was most efficient for conidiation. Whereas, some rice leaf media more effectively increased conidiation than panicle media except 0.10 g/ml panicle medium, and certain non-filtered tissue media were better than their filtered counterparts. Although the conidia induced in rice tissue media were smaller, they were able to germinate on potato sucrose agar medium and infect rice normally. The rice tissue medium is also workable in inducing conidia for conidiation-defective isolates. This method provides a foundation for the production of conidia by U. virens that will be widely applicable in pathogenicity testing as well as in genetic analyses for false smut resistance in rice cultivars.