Winemaking in Romania has a long-lasting history and traditions and its viticulture dates back centuries. The present work is focused on the development of wine yeast isolation and selection performed in different Romanian winemaking regions during past decades, presenting the advanement of the methods and techniques employed, correlated with the impact on wine quality improvement. Apart from the historical side of such work, the findings will reveal how scientific advancement in the country was correlated with worldwide research in the topic and influenced local wines’ typicity. To create an overall picture of the local specificities, the work refers to local grape varieties and the characteristics of the obtained wines by the use of local yeasts as compared to commercial ones. Numerous autochthonous strains of Saccharomyces were isolated from Romanian vineyards, of which several demonstrated strong oenological characteristics. Meanwhile, different non-Saccharomyces yeast strains were also isolated and are nowadays receiving the attention of researchers seeking to develop new wines according to wine market tendencies and to support wine’s national identity.