Rice false smut caused by Ustilaginoidea virens is one of the most important diseases of rice worldwide. Although its genome has been sequenced, to date there is no report on targeted gene deletion in U. virens and no molecular studies on genetic mechanisms regulating the infection processes of this destructive pathogen. In this study, we attempted to generate knockout mutants of the ortholog of yeast HOG1 MAP kinase gene in U. virens. One Uvhog1 deletion mutant was identified after screening over 600 hygromycin-resistant transformants generated by Agrobacterium tumefaciens mediated transformation. The Uvhog1 mutant was reduced in growth rate and conidiation but had increased sensitivities to SDS, Congo red, and hyperosmotic stress. Deletion of UvHOG1 resulted in reduced expression of the stress response-related genes UvATF1 and UvSKN7. In the Uvhog1 mutant, NaCl treatment failed to stimulate the accumulation of sorbitol and glycerol. In addition, the Uvhog1 mutant had reduced toxicity on shoot growth in rice seed germination assays. Overall, as the first report of targeted gene deletion mutant in U. virens, our results showed that UvHOG1 likely has conserved roles in regulating stress responses, hyphal growth, and possibly secondary metabolism.