Natural products have gained worldwide favor due to their sustainable and ayurvedic nature. In this study, rose containing anthocyanin was explored as a source of yellowish–pink natural colorant for wool dyeing using various solvents (aqueous, alkaline, acidic, and basified methanol). Dyeing of wool was performed by optimizing the following parameters: pH, temperature, time, and salt concentration. It was observed that aqueous extract with a pH of 3 employed at 80 °C for 35 min with the addition of 4 g/100 mL salt, acidic extract with a pH of 2 employed at 40 °C for 35 min with the addition of 2 g/100 mL salt, alkaline extract with a pH of 1 employed at 80 °C for 45 min with the addition of 3 g/100 mL salt, and methanolic extract with a pH of 2 employed at 80 °C for 45 min with the addition of 4 g/100 mL salt resulted in high tint (K/S) values. To improve the colorfastness properties, salts of iron (Fe+2), aluminum (Al+3), and tannic acid (Tn) were used as chemical mordant, whereas turmeric and pomegranate were used as biomordants. Iron (Fe+2) and pomegranate resulted in shades with good colorfastness characteristics. The plant extract and dyed fabrics were also evaluated against Gram-positive and Gram-negative bacteria Staphylococcus aureus and Escherichia coli, respectively, to observe their antibacterial potential. The results indicate that wool fabric dyed with naturally sourced rose petals can be used as valuable antibacterial fabric due to the presence of various bioactive compounds by dissipating the effect of allergy-causing synthetic dyed fabrics.