| Summary: | 碩士 === 大同大學 === 生物工程學系(所) === 94 === 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.
|
|---|
