ANTHOCYANIN ACCUMULATION AND GENE EXPRESSION ANALYSIS IN BLACK CARROT CALLI UNDER SALT AND SALICYLIC ACID STRESSES

Black carrot (Daucus carota ssp. sativus var. atrorubens Alef.), is an important crop plant from the Apiaceae family, rich in anthocyanins, which contributes to its antioxidant capacity. Anthocyanins are secondary metabolites give purple to red color to plants, aid to adapt plants to severe environmental conditions. The main purpose of this study is to investigate the effects of NaCl and salicylic acid (SA) stress conditions on anthocyanin accumulation and antioxidant activity of total anthocyanin extract from black carrot callus cultures. In this study, one month old calli elicited with 50, 100, 150, 200 mM NaCl and 50, 100, 200, 400 µM SA. qRT-PCR analyses were performed to examine gene expression in the phenylpropanoid pathway. The anthocyanin profiles of the extraction obtained from stressed purple carrot calli were determined through HPLC-DAD while antioxidant capacities were evaluated with DPPH and CUPRAC assays. Results indicated that SA application generally increased antioxidant capacity, with the highest activity observed particularly in calli treated with 150 mM NaCl on day 15. NaCl elicitation was more effective in inducing anthocyanin biosynthesis compared with SA. HPLC analysis demonstrated a notable increasement of anthocyanin accumulation in NaCl treated samples, dramatically on the 10th day with 200 mM NaCl. This study indicated that elicitation with NaCl and SA affected anthocyanin accumulation and antioxidant capacity in black carrot calli. Thus, it is suggested that optimized elicitation conditions can be used to provide an alternative approach to increase the amount of anthocyanins to utilize its valuable health-promoting properties. By exploring the molecular mechanisms of anthocyanin biosynthesis under NaCl and SA stress, future studies could shed light on understanding of plant defense, potentially contributing to the development of more resilient varieties. Keywords: Anthocyanin, Antioxidant Activity, Daucus carota ssp., Elicitation, Gene Expression, HPLC