Recently, the sustainability of agricultural production, influenced by intensive agricultural practices and climate change, has become increasingly important, particularly through the adoption of environmentally friendly methods. Researchers are showing growing interest in precision agriculture techniques, and sustainable farming practices are being actively developed. Phylloxera (Daktulosphaira vitifoliae Fitch) infestations have led to the widespread adoption of grafted vines as one of the most sustainable methods in viticulture. However, some grapevine rootstocks exhibit poor rooting and grafting success. To address this issue, synthetic chemicals are often used to enhance the rooting rate of cuttings. Nevertheless, the harmful effects of these chemicals on human health and the environment have increased interest in biostimulants in recent years. This study investigates the effects of thyme hydrolate on the rooting of grapevine cuttings. To this end, cuttings from the 41 B (V. vinifera x V. berlandieri) rootstock and the 'Narince' (V. vinifera L.) table grape variety were dipped in thyme hydrolate for three seconds before being planted in a rooting medium. The untreated cuttings served as the control group. Root number, root length (cm), fresh and dry root weights (g), shoot length (cm), and shoot diameter (mm) were measured. The results showed that thyme hydrolate application significantly increased root number, root length, and both fresh and dry root weights while reducing shoot diameter in both genotypes. The 'Narince' variety treated with thyme hydrolate exhibited the highest values for root length (24.67 cm), root number (12.67), root fresh weight (4.07 g), and root dry weight (0.70 g). In contrast, the control vines of the 'Narince' showed the greatest shoot length (25.67 cm) and shoot diameter (4.32 mm). Overall, the findings indicate that thyme hydrolate is a promising, environmentally friendly approach for enhancing root development in grapevine saplings.