The impact of non-Saccharomyces yeasts in fermentation has been investigated during recent years in order to assess the effect of using specific strains individually or in combinations in wine aroma (quality and complexity), as well as in alcohol content. The main purpose of this study was to explore the use of non-Saccharomyces yeasts, individually or in mixed culture with Saccharomyces cerevisiae for the vinification of must originating from the native white wine grape variety Savvatiano. Savvatiano is the most planted grape variety in Greece, cultivated predominantly in Central Greece, and consists the main grape used for the production of flavored wine Retsina. It produces wines with a low aromatic character but well balanced in terms of structure and taste when cultivated in high altitude. Grapes of Savvatiano were harvested during October 2020, were pressed, and the must after cold settlement was inoculated with a. Saccharomyces cereviasiae, b. Metschnikowia pulcherrima and c. mixed culture, in sequential inoculation (M. pulcherrima, followed by S. cereviasiae after 7 days). The progress of fermentations was monitored and the finished wines were analyzed for main wine parameters, as well as sensory attributes by a panel of experts. Wines fermented with S. cerevisiae were characterized by low aromatic intensity on the nose and lack of aromas on the palate, while wines fermented with M. pulcherrima exhibited low scores in general acceptance. However, sequential inoculation with M. pulcherrima, followed by S. cereviasiae produced a full-mouthed wine, characterized by aromatic complexity, citrusy aromas, lasting palate aromas, which exhibited high scores in general acceptance. The results of this study provide useful data in order to further explore the effect of mixed cultures use on fermentation of musts originating from native grape varieties with low aromatic profile.