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URN etd-0824106-091557 Statistics This thesis had been viewed 1722 times. Download 11 times. Author Hsien-shen Kuo Author's Email Address No Public. Department Bioengineering Year 2005 Semester 2 Degree Master Type of Document Master's Thesis Language English Page Count 74 Title Production of resveratrol by cell culture of Vitis thunbergii Sieb.et Zucc. Keyword Vitis thunbergii resveratrol plant cell culture plant cell culture resveratrol Vitis thunbergii Abstract The callus of Vitis thunbergii Sieb. et Zucc. was subcultured bimonthly for three years (long-term cell) and was used to establish suspension culture and examine the content of resveratrol in cells. Biomass of long-term cell increased nearly 3.5 times every 18 days cultured in 1/2 MS medium supplemented with 1.86 mg/L NAA and 0.22 mg/L BA. Biotic factors (fungi hypha and yeast elicitor) and abiotic factors (jasmonic acid , methyl jasmonate , salicylic acid, phenylalanine, cinnamic acid, UV ) were used to treat the cells to enhance accumulation of resveratrol. The resveratrol content of cells reached to 267.29±70.59 mg/kg DW (the control was 21.79±8.69 mg/kg DW) when cell cultured in modified MS medium contained autoclaved hypha of Botyis cinerea, but the cells dead quickly. Methyl jasmonate at 6 mg/L in medium, resveratrol content boosted to 198.59±66.95 mg/kg DW and the fresh weight of biomass to 12.57 g (original inoculum was 3 g) for 18 days culture. Cells treated with UV (power:30W, time:30 minutes) proliferated normally and the resveratrol content in cells was 110.18±34.72 mg/ kg DW. Addition of high dose of sucrose (150 g/L) cause resveratrol accumulation (216.08±65.18 mg/kg DW) but cells eventually died.
The fast growing callus induced from stem and petiole were cultured on modified MS medium with 1.5 mg/L NAA+ 0.5 mg/L BA (new callus). After three subcultures, the content of resveratrol in new callus was 3616.36 mg/kg DW. The growth rate of long-term cell line was 3.5 folds after 18 days culture, but the content of resveratrol was low (21.79 mg/kg DW). By adding elicitor to medium, the content of resveratrol in long-term cells only up to 267.29 mg/kg DW. It was expressed that the productivity of resveratrol in long-term cell line was fast decrease.
Advisor Committee Chin-wen Ho - advisor
Kow-jen Duan - co-chair
Wei-chin Chang - co-chair
Files Date of Defense 2006-07-07 Date of Submission 2006-08-24