The increasing demand for natural bioactive compounds in health and functional nutrition motivates the development of sustainable biotechnological approaches. This study aimed to optimize cultivation and genetic transformation strategies in Brassica oleracea var. capitata f. rubra (purple cabbage) to enhance the production of anthocyanins, phenolic pigments with recognized antioxidant and anti-inflammatory effects. Seed disinfection and germination protocols were refined, followed by in vitro cultivation using Murashige and Skoog media under complete and nitrate-free conditions. The impact of biotic elicitors, including methyl jasmonate and salicylic acid, was evaluated. Additionally, genetic transformation via Agrobacterium rhizogenes was performed to induce hairy roots expressing the transcription factor AtMYB12 and the reporter gene GUS. Results indicated that cold stratification combined with gibberellic acid improved germination efficiency. Seedlings grown in nitrate-free medium accumulated higher anthocyanin levels with enhanced antioxidant and anti-inflammatory activities. Transformed roots expressing AtMYB12 in complete medium exhibited further increased anthocyanin content and bioactivity. Statistical analysis highlighted the significant effect of nitrate deprivation on seedlings and the pivotal role of AtMYB12 expression in transformed roots. These findings demonstrate that integrating optimized culture conditions with genetic engineering can effectively boost the production of bioactive anthocyanins in purple cabbage, offering sustainable alternatives for generating functional compounds with potential applications in food, pharmaceutical, and cosmetic industries. Moreover, this approach contributes to the conservation and valorization of plant genetic resources.

Keywords: Brassica, Anthocyanins, plant biofactory, genetic transformation