Abstract Background: Biopriming as a new technique of seed treatment involves the application of beneficial microorgan‑ isms on the seed surface to stimulate seed germination, plant growth, and protect the seed from soil and seed‑borne pathogens. The present investigation was carried out on seed germination, seedling vigor and biochemical traits of one of the most important vegetable crops (Tomato, Solanum lycopersicum L.). The treatments comprised viz. T1: Non primed seeds (Control), T2: Hydropriming, T3: Biopriming with C‑phycocyanin (C‑PC) (Spirulina platensis extract), T4: Biopriming with Trichoderma asperellum, T5: Biopriming with T. viride, T6: Biopriming with Beauveria bassiana. Results: Extraction and purification of C‑phycocyanin (C‑PC) from the dry S. platensis powder using various methods was performed. The purity after dialyses was 0.49 and its ultimate purity (A620/A280) after ion‑exchange chromatogra‑ phy was 4.64. The results on tomato seedlings revealed that the maximum germination percentage (100%), germina‑ tion index (15.46 and 15.12), seedling length (10.67 cm), seedling dry weight (1.73 and 1.97 mg) and seedling length vigor index (1066.7) were recorded for tomato biopriming with T. viride, and B. bassiana (T5 and T6). Moreover, the quantitative estimation of total carbohydrates and total free amino acids contents in bioprimed tomato seedlings indicated a significantly higher amount with T. viride, followed by those bioprimed with T. asperellum, B. bassiana and C‑PC extract. Conclusion: Thus, our results indicated that biopriming of tomato seeds with beneficial fungal inoculants and C‑PC was very effective. The most operative biostimulants were those bioprimed with T. viride and B. bassiana compared to other biostimulants (T. asperellum and C‑PC). Therefore, to ensure sustainable agriculture, this study offers new possibilities for the biopriming application as an alternative and ecological management strategy to chemical treatment and provides a valuable basis for improving seed germination.