DEVELOPMENT OF IN VITRO PROTOCOLS TO ENHANCE SECONDARY METABOLITE PRODUCTION FROM TURMERIC (\u3ci\u3eCURCUMA LONGA\u3c/i\u3e L.)

Plant and organ culture can readily be applied to the field of medicinal plant and secondary metabolite production. Many studies that have been conducted attest to the utility of in vitro methods for preservation of genetic diversity, propagation of clonal individuals for use in commercial field planting, and production of metabolites directly from culture. Experiments have also shown that tissue culture is an important tool for enhancement of medicinal compound production utilizing various methods including elicitation. This thesis focuses on production of metabolites using an in vitro system. An overview in the form of a literature review is provided, which details the many ways that tissue culture has been applied to medicinal plants. Two major concerns are addressed experimentally, quantity of biomass and concentration of compounds in that biomass. Quantity of biomass was evaluated in a time course factorial experiment employing three different concentrations each of methyl jasmonate (MeJa) and benzyladenine (BA). MeJa negatively impacted biomass accumulation while BA had a positive impact on biomass accumulation. Sugar use was found to directly correlate with dry weight production, and a system for long-term growth of plant materials was developed. Concentrations of compounds in the biomass were explored by observing effects of media additions of proline, a natural source of proline (Gropro), MeJa, phenylalanine, chitosan, and nitrogen stress. MeJa and phenylalanine were utilized in a factorial experiment, and both compounds reduced biomass and secondary metabolite production. Proline, Gropro, MeJa, chitosan, and nitrogen stress were evaluated in comparison with a control in a separate experiment. Only one treatment, nitrogen stress, was successful in upregulating metabolite production. Nitrogen stress was found to increase total phenolic production. It was also successful in causing a nearly significant (p = 0.1207) increase in antioxidant activity from extracts. Sugar and nutrient status are critical factors relating to our main goals, enhancement of biomass production and enhancement of chemical concentration. As a result of these two bodies of work, we conclude that future experiments should evaluate nutrient stressed plants in elicitation studies where sugar is optimized for maximum biomass production. Other upregulation methods would be employed in this new system to further enhance chemical production