Date of Award
Master of Science (MS)
Plant and Environmental Science
Knap , Halina
Bridges , William
Murashige and Skoog 1962 performed one-factor-at-a-time (OFAT) experimentation in order to regenerate tobacco callus on semi-solid agar medium. This work became an established medium for tissue culture micropropagation and experimentation. Micropropagation is done in niche markets with herbaceous perennials, among other crops, and the optimization of various inputs to produce maximal responses is a necessary step towards process development. This current study of macronutrient factors simultaneously altered media volume, amount of tissue (plants per vessel), sucrose, nitrogen (as NO3- and NH4+ ions), and K+ in a d-optimal design space with only 55 experimental units (including 5 true replicates). The first study examined these macronutrients in a micronutrient limited environment, and probed further areas in the design space for exploration. The second study, at full MS meso- and micronutrient values, identified P and Mg to be deficient in standard formulations when compared with field-grown plantlets of turmeric, while identifying differences in the definition of plantlet quality. Plantlet quality was defined in three very distinct ways. These three optimization choices were demonstrated to have very different optima as defined in this experiment. First: multiplication is maximal with low plantlet density (3 plantlets per vessel), high media volume (45 ml), and 4% sucrose in the vessel. Secondly: the number of new plants produced per vessel was highest when the most plants were put in a vessel, at the highest media volume, and highest sucrose % tested. Lastly, those individual plantlets transferred to the greenhouse (100% plantlet survival) which grew the most (via fresh mass gains) were the most massive (fresh mass) came from vessels with: 3 plantlets per vessel, containing no NH4+ (all K+), 45 ml media volume (the highest amounts of media components holding concentrations constant), and the lowest concentration of sucrose tested (1.5%). These two experiments result in separation of optima demonstrating the need for differing tissue culture medium formulations that are dependent upon the process of interest, while identifying possible areas of future work necessary for in vitro nutritive media formulations in turmeric (Curcuma longa L.), an important medicinal herbaceous perennial.
Halloran, Sean, "SIMULTANEOUS OPTIMIZATION OF MACRONUTRIENT FACTORS IN PLANT TISSUE CULTURE USING TURMERIC (Curcuma longa L.) AS A MODEL" (2011). All Theses. 1125.