Suspension Culture and Alkaloid Content of Pinelliae Rhizoma Tuber Under Stimulation of Salicylic Acid / 中国实验方剂学杂志

ABSTRACT

Objective: To study the suspension culture of tuber and its alkaloid content based on the stimulation of salicylic acid. Method: The tubers of Pinelliae Rhizoma in suspension tube were treated with different concentrations of exogenous salicylic acid to analyze the growth status. The content of alkaloids in tuber was detected by HPLC. Test conditionschromatographic column for Agilent Eclipse plus C18 column (4.6 mm×250 mm,5 μm),mobile phase of acetonitrile water(496),the column temperature was maintained at 35℃,detection wavelength for inosine 250 nm,guanosine 260 nm,volume flow rate 1.0 mL·min-1. Result: The results showed that the exogenous salicylic acid had a certain effect on the growth of suspension tuber of Pinelliae Rhizoma. When the salicylic acid concentration was 150 μmol·L-1,the culture lasted for 25 days and the fresh weight reached the maximum value of 7.483 8 g. It also accumulates a certain amount of alkaloids. The linear range of guanosine was 0.03-0.45 μg (R2=0.999 6). After 10-days cultivatation in the salicylic acid concentration of 50 μmol·L-1,guanosine content of Pinelliae Rhizoma tubers reached a maximum of 1.353 3 mg·g-1. The linear range of inosine 0.003-0.045 μg (R2=0.999 5). When the salicylic acid concentration was 200 μmol·L-1,cultured for 30 days,the content of inosine in Pinelliae Rhizoma tubers reached the maximum value of 0.149 8 mg·g-1. Conclusion: The results of this experiment provide a reference for the study of tissue culture and rapid propagation of Pinelliae Rhizoma tubers and regulation of alkaloids,which are of great significance for the development of Pinelliae Rhizoma industry.