Organic materials are increasingly being used in various applications, including industrial and agricultural production, due to their fewer side effects compared to chemical substances. This study was conducted in 2014 in two separate experiments using a completely randomized design to investigate the effect of pine wood vinegar on the growth and physiological characteristics of basil with six treatments (0, 1250, 2000, 2500, 3333, and 5000 mg/L) with four replications. The first experiment was conducted in the laboratory on basil seeds as a priming treatment, while the second experiment was conducted in a greenhouse at the Agricultural Faculty of Isfahan University of Technology. The application of pine wood vinegar on seeds at a concentration of 1250 mg/L increased the germination percentage, while a concentration of 2500 mg/L accelerated the germination process. In the second experiment, the highest root volume and length were observed in the treatment with 3333 mg/L, while the highest root surface area was observed in the treatment with 5000 mg/L. The concentration of nitrogen was highest in the treatment with 3333 mg/L, potassium concentration was highest in the treatment with 5000 mg/L, and calcium and iron concentrations were highest in the treatment with 1250 mg/L compared to other treatments. Chlorophyll fluorescence intensity, SPAD index (chlorophyll content), ion leakage, and chlorophyll ((A)) were highest in the treatment with 3333 mg/L. Based on these results from Experiment Two, basil plants treated with pine wood vinegar at a concentration of 3333 mg/L showed the greatest increase in growth indices. Therefore, this concentration can be recommended for use.