The current list of fungi from Colombia updated in the present review contains a total of 7619 species. The Ascomycota appears as the most diverse group, with 4818 species, followed by the Basidiomycota, with 2555 species. Despite this, we presume that the actual fungal diversity in Colombia could amount to between 105,600 and 300,000 species. Fungi represent an underestimated resource, indispensable for human well-being. Even though the current knowledge on potential applications of Colombian fungi is still limited, the number of studies on areas such as natural products discovery, biological control, and food and beverages, among other biotechnological applications, are increasing. With the current review, we aim to present a comprehensive update on the fungal diversity in Colombia and its potential applications. Colombia’s native fungal biodiversity holds much potential within the country’s current social-economical context, and the future must ensure efforts to preserve both the biodiversity and the untapped resources of the fungi in Colombia, which in alignment with the Sustainable Development Goals (SDGs) might result in new bioeconomy avenues for the country.