Tryptophan, as a signal molecule, mediates many biotic and environmental stress-induced physiological responses in plants. Therefore, an experiment was conducted to evaluate the effect of tryptophan seed treatment in response to cadmium stress (0, 0.15, and 0.25 mM) in sunflower plants. Different growth and biochemical parameters were determined to compare the efficiency of the treatment agent. The results showed that cadmium stress reduced the growth attributes, including root and shoot length, dry and fresh weight, rate of seed germination, and the number of leaves. Cadmium stress also significantly reduced the contents of chlorophyll a, b, and total chlorophyll, carotenoid contents, phenolics, flavonoids, anthocyanin, and ascorbic acid. Whereas cadmium stress (0.15 and 0.25 mM) enhanced the concentrations of malondialdehyde (45.24% and 53.06%), hydrogen peroxide (−11.07% and 5.86%), and soluble sugars (28.05% and 50.34%) compared to the control. Tryptophan treatment decreased the effect of Cd stress by minimizing lipid peroxidation. Seed treatment with tryptophan under cadmium stress improved the root (19.40%) and shoot length (38.14%), root (41.90%) and shoot fresh weight (13.58%), seed germination ability (13.79%), average leaf area (24.07%), chlorophyll b (51.35%), total chlorophyll (20.04%), carotenoids (43.37%), total phenolic (1.47%), flavonoids (19.02%), anthocyanin (26.57%), ascorbic acid (4%), and total soluble proteins (12.32%) compared with control conditions. Overall, the tryptophan seed treatment showed positive effects on sunflower plants’ growth and stress tolerance, highlighting its potential as a sustainable approach to improve crop performance.