植醇与寡糖对红花悬浮培养细胞的生长及α-生育酚积累的效应

Effects of Phytol and Oligosaccharide on Cell Growth and α-Tocopherol Production in Cell Suspension Culture of Carthamus tinctorius

在红花细胞悬浮培养基中加入α-生育酚的前体植醇能显著增加α-生育酚的积累,较高浓度的植醇(50～100 ppm)能同时促进细胞的生长。在培养基中加入抗氧剂角鲨烯也能提高α-生育酚的积累。诱导因子寡糖对细胞的生长和α-生育酚含量均有提高作用,其中以寡糖F_(11)效果较佳。悬浮细胞培养基中同时加入15ppm的寡糖F_(11),75ppm的植醇和100 ppm的角鲨烯,细胞生长速率可提高23.81％,α-生育酚含量及产率分别比对照提高2.8倍和3.6倍。

For the purpose of increasing α-tocopherol content in the suspension culture cells of Carthamus tinctorius, the effects of phytol and oligosaccharide on cell growth and α-tocopherol production in cell suspension culture were studied. The inoculum of cell suspension cultures was obtained from the callus line induced from the hypocotyl of G. tinctorius and successively subcultured over 15 generations. MS medium supplemented with 1 ppm 2, 4-D, 0. 1ppm KT and 0.1% CA (casamino acid.) was used as suspension medium. The cells to be cultured were collected and freeze-dried, then the cell growth rate was examined. The α-tocopherol content was determined by HPLC method. Dry samples were extracted with petroleum ether. The extracts were evaporated in vacuo, then determined by HPLC on Zorbax ODS column (4.6×250 mm, Shimadzu) and eluted with methanol, α-tocopherol was monitored at 290 nm UV wavelength and its content calculated using an external standard. The results indicated that the addition of phytol increased the α-toeopherol content, and higher concentrations (50～100 ppm) could increase cell growth rate. Adding squalene, an antioxidant, to the medium prevented α-tocopherol from being oxidized and thereby effectively increased its accumulation. The test on elicitors from plants as Dendrobium candium and Panax ginseng proved that both cell growth rate and α-tocopherol production were promoted by elieiting with oligosaccharides, of which oligosaceharide F_(11) was the most effective. The test sunder optimum culture conditions showed that cell growth rate was increased by 23.81%, and the α-tocopherol content and yield were increased by 2.8 and 3.6-fold, respectively, when the cells were incubated in a liquid medium supplemented with 15ppm oligosaccharide F_(11), 75ppm phytol and 100ppm squalene. It was shown that the products of secondary metabolism in plant cell cultures could be increased by regulating the culture condition.