| Summary: | 碩士 === 國立屏東科技大學 === 生物科技系所 === 102 === A gradual substitution of fossil energy to renewable energy is a global trend, not only it resolves and increases the critical energy shortage, but also crisis of greenhouse effect. Jatropha curcas is an inedible crop, the seeds containing about 40% lipids can be used as biodiesel. The aim of the current research project was to apply a nonconventional bioprocess for in vitro biomass and oil production of Jatropha cells. The seeds of Jatropha curcas were used for aseptic seeding. The methods of callus induction, suspension cell culture and embryogenesis of Jatropha were established. The best inducing rate of buds was achieved by adding 1.5 mg/L BA and 0.5mg/L IBA in MS. The suspension cells were cultured in light and dark conditions. Once friable callus was obtained, and cell suspension cultures were established. The cell viability, fatty acid content, and characteristics were used to select the most promising cell line according to its fatty acid profile and ability to grow and develop under in vitro conditions. Nile red staining showed that suspension cells in light accumulate more lipid than in dark. To characterize the metabolic mechanism of lipid accumulation in Jatropha suspension culture cells, comparative proteomic technique was employed to profile protein changes during different growth condition. The fatty acids were extracted and run through thin layer chromatography (TLC), and the results showed that suspension cells in light and dark accumulated different fatty acids and their derivatives. Differences growth conditions of Jatropha were related to their fatty acid profiles, oil content. We used comparative proteomics to understand the metabolic mechanism of lipid. The protein were identified by LC/MS. Peroxidase 68 OS、Hypothetical protein CHLREDRAFT_150509、glutathione S-transferase omega、translationally controlled tumor protein、annexin-like protein、Glyceraldehyde-3-phosphate dehydrogenase、major allergen Pru ar were found in dark. Major allergen Pru ar was found more in dark than light, and it was associated with plant pathogenic. It is helpful in plant lipid synthesis and may make the Jatropha suspension cultured cells can be as bioreactor to produce biodiesel. It can reduce cost in the production process and improve the properties of biodiesel. Also improving the understanding of fatty acids biosynthesis in higher plants.
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