Rosemary Rosmarinus officinalis is a member of the Lamiaceae. Rosmarinic acid (RA) is a very strong antioxidant produced in the chloroplast, and used to protect plant tissues against oxidative stress. A number of investigations showed that the sucrose concentration in the callus growing medium greatly influenced the production of secondary metabolites of the phenylpropanoid pathway such as RA. The aim of this study was to test the effect of elevated sucrose concentrations (2%, 3%, 4%, 5%, and 6% sucrose) and the effect of light and dark treatments on the production of RA in the callus of five different genotypes. The genotypes were Majorca, Rosmarinus officinalis, Pine Scented, Madeline Hill, and APR. It was found that the dark treatment produces more RA than the light treatment in all genotypes, and in all sucrose concentrations. The RA concentration increased with increasing the sucrose concentration from 2%—reaching the highest concentration at 4% and 5% in most genotypes. The RA concentration declined again at 6% sucrose in all genotypes. We concluded that for the extraction of RA from rosemary callus it is preferred to be produced in the dark—this will save energy and will produce more RA than the light treatment. Also it is preferred to use sucrose concentration at 4% for genotypes Rosmarinus officinalis, Pine Scented, and APR; and 3% sucrose for genotype Madeline Hill in the dark condition. While for the light condition, it is preferred to use 5% sucrose with genotypes Majorca, Rosmarinus officinalis, Pine Scented, and Madeline Hill; and 4% sucrose for genotype APR.
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