Studies On The Adventitious Root Culture Of Salvia Miltiorrhiza Bge.

In this project, we studied adventitious root culture of Salvia miltiorrhiza Bge.. The callus was induced and proliferated by using young leaves. Effects of different physical and chemical factors on adventitious root suspension culture were investigated, which provided scientific basis and technical support for adventitious root culture in big scale and commercial metabolite yields. The results are listed as follows:(1) MS medium with a combination of IBA 2 mg·L-1 and KT 0.2 mg·L-1 was suitable for the induction and subculture of adventitious roots.(2) Adventitious roots grew better under high salt intensity while secondary metabolite biosynthesis was accelerated under low salt intensity in MS basal medium.(3) Compared with dark culture, light culture was more favorable for root growth and secondary metabolite biosynthesis.(4) High root multiplication rates were obtained at the pH value of 4.5, 6.5 and 7.0, while the maximum productions of tanshinone IIA (10.29μg·g-1) and protpcatechuic aldehyde (251.56μg·g-1) were achieved at the pH value of 5.5 and 6.0, respectively.(5) Inoculum size of 2.5% was favorable for the root culture, under which root multiplication rate(2.88-fold),contents of tanshinone IIA(28.74μg·g-1) and protocatechuic aldehyde(295.11μg·g-1)were obtained.(6) Carbon, nitrogen and phosphate sources were necessary for the adventitious root culture. High root multiplication rate was achieved at sucrose level of 30 g·L-1 after twtnty-day culture, while 60 g·L-1 sucrose and low level sucrose were favorable for tanshinone IIA and protocatechuic aldehyde formation, respectively. The highest root yield and tanshinone IIA content on day 25 were obtained by intermittent sugar feeding during cultivation, and the production of roots and tanshinone IIA increased 2.3- and 2.4-fold, respectively, compared with those of control. The maxium root growth rate, contents of tanshinone IIA and protocatechuic aldehyde were obtained while NH4+/ NO3- was 1∶4,1∶4 and 1∶1, respectively, when total nitrogen source was kept 60 mmol·L-1. Changing concentrations of KH2PO4 was more favorable for root growth, whereas high concentration of KH2PO4 inhibited tanshinone IIA biosynthesis.(7) Natural Fe²⁺ and Mn²⁺, 2.0-fold Cu²⁺ and 1.5-fold Mg²⁺ improved root growth;...