The Suspension Culture Of Compact Callus Aggregates Of Rhodiola Sachalinenis And The Induction Of Salidroside

Rhodio/a sachalinensis A. Bor is a kind of precious herb of the traditional Chinese medicine. Its root and rhizome contains salidroside, which has been shown to possess the medical functions such as resisting anoxia, microwave radiation and fatigue. in order to establish the differentiated callus suspension culture of Rh. Sachahnen.cis with high salidroside production. the induction of callus of Rh. sachalinensis, the regulation of the morphas of the callus, the establishment of the compact callus aggregates (CCA) suspension culture. the characteristics of the growth of CCA suspension culture and the accumulation of salidroside. the improvement of the culture media and culture conditions, the strategies of elicitation and precursors feeding, and the biosynthesis pathway of salidroside were studied in this experiments. 7 types of callus were induced from the cotyledon, the spire, the caulicle and the root of Rh. Sachalinen.cis. The induction of callus from the cotyledon is easiest, from spire is easier and from the root is very difficult. By regulating the morphas of the callus, the preIirninaril~ differentiated compact callus aggregates (CCA) with higher saLidroside content and higher giowth rate was obtained. The growth c~ cle of the CCA suspension culture is about 25 ?--3(1 days. The period from 0 to 1 5 days after subculture is the exponential stage of the gro~vth of the culture. From 15 to 20 days after subculture. the growth of the culture begins to decrease. The maximum of the accumulation of salidroside appears on the day 3 ?? after subculture. The basic MS liquid media with 5 nig/L BA and 2.5 mg/L IBA benefit the growth of the culture. Exogenous 2,4-D inhibited the growth of the culture greatly but stimulated the accumulation of salidroside significantly. If the basic MS medium is the standard. maintaining the quantity of total nitrogen and pH value and increasing the quantity of total sugar to 45 g/L improved the growth of the culture but inhibited the accumulation of salidroside. While. decreasing quantity of both total nitrogen and total sugar to half level and decreasing pH ~ alue to 4.2 separately improved the accumulation of salidroside but inhibited the growth of the culture. Low temperature at 15?C and low shaking speed with 100 rprn/miii benefit the growth of the culture, while temperature at 24~?C and high shaking speed with 150 rpm/rn in benefit the accumulation of salidroside. The ideal induction efficiency of UV irradiation (200 W/m2) on improvement of salidroside production of the culture is carried out for 20 mm at day 15 after subculture. The induction efficiency of the extract of the root of Rhododendron chrysanthum is higher than that of Saussurea alpicola. The improvement efficiency of possible precursor of salidroside such as Iv I / tvrosol, phenylalanine, salicylic acid, tryptophan, and tyrosine become less. ~Two-step?culture method is suggested to improve the salidroside production in mass CCA suspension culture of Rh. Sachalinensis. In order to gain the maximum biomass of the culture first. the CCA is cultivated in the growth-medium for 15~?O days. Then the CCA is transferred into production-medium to incubate for 3 5 days before cultivated. The growth-medium has the same amount of total nitrogen, the same pH value and 1.5 folds of total sugar as the basic MS medium with 5 mg/L BA and 2.5 mg...