Cell Suspension Culture Of Strobilanthes Cusia (Nees)O. Ktze And Extraction And Analysis Of Its Indigotin And Indirubin

In this study, the indigotin and indirubin were obtained from the suspension-cells cultured in the established cell suspension culture system with the calli induced from the seed of Strobilanthes cusia (Nees) O. Ktze. On the study of influence of different combinations of hormone, nitrogen source, phosphorus source, carbon source, inostol and the cell inculated amount on the Strobilanthes cusia (Nees) O. Ktze' cell growth and synthesization of the indigotin and indirubin, the dynamic curves were established in order to the highest yield of the indigotin and indirubin though cell suspension of Strobilanthes cusia (Nees) O. Ktze.1. Plenty of more uniformly and quickly growing callus was harvested from the MS medium with 0.2mg·L^-12,4-D ,1.0mg·L^-1NAA ,1.0mg·L(-1)BA.2. The HPLC system of the indigotin and indirubin : C18(Waters 2695-2996), CH₃OH-H₂O(90:10), 0.5mL·min^-1, 290 nm. At about 20 days, the content of the indigotin and indirubin was 1.35 mg(DW)·g^-1, 0.42 mg(DW)·g^-1 respectively.3. After optimized culture condition for cell suspension culture,the results showed that the best culture condition was the modified MS medium with 255 mg·L^-1 KH₂PO₄, 2475 mg·L^-1 NH₄NO₃, 150 mg·L^-1 inositol, 50g·L^-2 sugar and 0.4mg·L^-12,4-D, 3.0mg·L^-1NAA, 0.1mg·L^-1KT, 0.2mg·L^-1BA, and added 0.04 g(DW) cell in 20mL liquid medium.4. Under the optimal culture condition,the dynamic curves between the Strobilanthes cusia(Nees ) O. Ktze's cell growth and synthesization of the indigotin and indirubin showed that the Strobilanthes cusia (Nees ) O. Ktze's cell growth reached to the top at the 24^th day and the biomass was 21.39 g(DW)·L^-1, and the content of the indigotin and indirubin reached to the top at the 27^th, at 28.44 mg(DW)·L^-1,8.78 mg(DW)·L^-1.