Studies On Establishments Of Callus And Suspension-cultured Cell Lines And Analyses Of Important Secondary Metabolites Of Rhodiola Crenulata

As a main efficient component of Rhodiola plants, salidroside is a kind of promising environmental acclimation medicine, with such specific medicinal properties as anti-fatigue, anti-decrepit, anti-microwave radiation, anti-virus, and anti-tumor. It plays an important role especially in military, aerospace, sports and healthcare medicines. In order to resolve the problem of a shortage of Rhodiola's resources, the suspension system of Rhodiola crenulata was built by the technique of plant tissure culture and cell culture in the present research. Take the suspension cell as material, some data related with the metabolism of salidroside were surveyed, also salidroside and tyrosol in the suspension system were analyzed by HPLC.Firstly, the study in the paper built the aseptic culture system of Rhodiola crenulata by taking the seeds as explants and the method of 70% ethanol sterilizing 30 seconds together with 9% NaClO 10 minutes. And then the callus of different parts of the plant was induced. The results showed that the younger leaves or petioles were easier to produce callus. The optimal culture medium to induce callus is 1/2MS + 6-BA 0.5 mg/L + NAA 3.0mg/L. Next, the steady callus was transferred into the liquid culture medium added with TDZ 0.5mg/L and NAA 0.5mg/L to build the suspension system. In this system, at room temperature, the biomass of callus propagated in a way of 'S' growth curve. And the PAL activity increased firstly, and then declined by time pasted. While theβ-D-glucosidase activity changed similarly with PAL, furthermore, both maximum appeared the same day. In a growth cycle, the glucose content went up first, had a ascend trend next, and then declined. In the prior period, amylose content had a steady trend, and then sharp increase and dropped. The optimal technics of extracting salidroside and tyrosol was as following: dip twice the callus powder in petroleum ether, each time for 30 minutes; ultrasonic extract once with 75% ethanol and the time of 30 minutes. The extracting rates of salidroside and tyrosol is 29.037mg/g and 15.633mg/g respectively.