Optimizing Tissue Culture of Anoectochilus roxburghii for a Maximized Content on Effe

Conditions of tissue culture for Anoectochilus roxburghii were optimized to maximize the content of effective substances in the plantlets. The tissues were cultivated in test tubes containing a basal medium of MS+6-BA 2.0 mg·L^-1+NAA 0.2 mg·L^-1+KT 1.5 mg·L^-1+agar 7 g·L^-1+glucose 25 g·L^-1. Effects of various organic additives, activated carbon additions and light exposures on the accumulation of polysaccharides and total flavonoids during the growth periods of the plantlets were studied using a L9 (3⁴) orthogonal experimental design. It was found that the treatment effects were in the order of:light > additives > activated carbon. The average increases on the target substances by the blue light exposure were significantly higher than those by the white or red light (P < 0.05). The increases by adding banana mud to the basal medium were significantly greater than the addition of either apple juice or carrot juice (P < 0.05). And, the gains by inclusion of 1.0 g·L^-1 of activated carbon in the medium were significantly higher than those at 1.5 g·L^-1 level (P < 0.05). Thus, it was concluded that an optimal production of polysaccharides and flavonoids in the plantlets could be achieved by a combination of the following ingredients in the medium under blue light:MS+6-BA 2.0 mg·L^-1+NAA 0.2 mg·L^-1+ KT1.5 mg·L^-1+agar 7 g·L^-1+glucose 25 g·L^-1+banana puree 100 g·L^-1+activated carbon 1.0 g·L^-1.