Research On Suspension Cell Culture Of Psammosilene Tunicoides

Psammosilene tunicoides is a kind of precious and endangered medicinal plant distributed exclusively in the southwest of China, which has highly economic value. Production of bioactive secondary metabolites can effectively overcome the problem of insufficient supplement of P. tunicoides herb and wild resource protecting by plant cell suspension culture.The normal P.tunicoides callus and P.tunicoides hairy root callus was used as materials in this paper. The system of plant cell suspension culture of P.tunicoides callus and P.tunicoides hairy root callus was established and the basic culture medium, hormone combination, biomass, growth curve and saponins content of two kinds of suspension cell system were researched. The results indicated that MS and 1/2MS medium were respectively suitable for normal suspension culture cells and P. tunicoides suspension culture of hairy root-derived cells. Both the optimal initial quantity were about 60 g·L-1 (FW). The best hormone combination of normal suspension cells culture was 1.0 mg·L-1 12,4-D+1.0 mg·L-1 6-BA+0.5 mg·L-1 NAA and the other one was 1.5 mg·L-1 2,4-D+0.5mg·L-1 6-BA+0.25 mg·L-1 NAA+0.1 mg·L-1 KT. The maximum biomass was 12.35 g·L-1(DW) achieved when normal suspension cell was cultured 24 d and the growth rate was 0.43 g·L-1·d-1.The maximum biomass of hairy root suspension cell was 14.59 g·L-1(DW) achieved and the growth rate was 0.72 g·L-1·d-1 after culturing 20 d. The saponins contents of normal and hairy root suspension cells were 0.45 and 0.92%, respectively. The basic medium, hormone formula, biomass, growth cycle of both suspension culture were compared. The hairy root-derived cells were more suitable for establishing cell suspension culture as materials because of its fast growth, short training period, and high levels of secondary metabolites.The nutrient consumption of nitrogen, carbon and phosphorus source, the proportion of nitrate nitrogen and ammonium nitrogen and the effect of elicitor on cell growth and secondary metabolism was investigated. The consumption of nitrogen, carbon and phosphorus source was synchronous with the growth of suspended cells and the ammonium nitrate consumed faster than nitrate nitrogen. When the ratio of NO3--N/NH4+-N was 2:1 in the culture media could achieve prefect result. The effect was better when the concentration of salicylic acid and chitin were 0.4 mmol·L-1 and 0.1%, respectively and the accumulation of saponin was 0.99%.