In this study, the yeast microbiota of naturally fermented black olives made from cv. Gemlik, grown in three different districts of the Cukurova region of Turkey, were investigated. Fermentations were conducted for 180days in three different brines, including NaCl 10% w/v, NaCl 8% w/v and NaCl 8% w/v added with glucose 0.5%. In total, 223 yeasts were isolated and then identified by PCR-RFLP analysis of the 5.8S ITS rRNA region and sequence information for the D1/D2 domains of the 26S rRNA gene. A broad range of yeast biodiversity was identified, including eight genera and nine species. Candida boidinii (41%), Wickerhamomyces anomalus (32%) and Saccharomyces sp. (18%) were predominant yeasts throughout the fermentations. To a lesser extent, the other species, Candida aaseri, Meyerozyma sp., Zygoascus hellenicus, Pichia kudriavzevii, Schwanniomyces etchellsii and Candida atlantica were also members of the olive-fermenting microbiota. In Tarsus and Bahce districts C. boidinii and in Serinyol district Saccharomyces sp. were the most frequently identified species. W. anomalus was the most frequently isolated species (by 48% of total yeasts) in NaCl 10% brines. C. boidinii was the most dominant species in the brines, including NaCl 8% and NaCl 8%+glucose 0.5%, with frequencies of 42% and 61%, respectively. At the end of the 180days of fermentation, total acidity values of the brines were in the range 1.04-8.1g/l lactic acid. Copyright (c) 2016 John Wiley & Sons, Ltd.