During traditional hardaliye production by fermentation from grape juice, mustard seeds are insufficient to inhibit yeast activities and alcohol formation. Chemical preservatives are used for the production of hardaliye of standard quality in traditional and industrial production. Today, consumers prefer natural products that do not contain chemical preservatives and additives. For this reason, in this study, hardaliye production was carried out with two different methods as back-slopping and addition of kombucha mushrooms. These methods were tried for the first time on hardaliye. Fermentation continued for 7 days and storage for 14 days. During fermentation, pH and reducing sugar, L* and a* values of samples decreased, while phenolic substance, viscosity and b* values increased. The pH continued to drop during storage. No significant changes were observed in reducing sugar values. The decrease in the amount of phenolic substances and the increase in viscosity and L* values of the kombucha hardaliye samples (KH) during storage were found to be statistically significant (p<0.05). The titratable acidity increase was greater in the back-slopping hardaliye (BH) sample compared to the KH sample. While 5.5% alcohol formation was observed in the BH sample on the 7th day of fermentation, no alcohol formation was detected in the KH sample. At the end of fermentation and storage, Total mesophilic aerobic bacteria (TMAB), yeast, Lactobacillus spp. and lactic streptococci numbers were found to be higher in BH sample than in KH sample. According to the results of the research, thanks to the metabolic activities of the kombucha mushroom microorganisms and their symbiotic association, natural fermentation takes place without the use of any preservative chemicals and more reliable hardaliye production is provided compared to back-slopping hardaliye.