North China Rhubarb Cell Suspension Culture And Antioxidant Activity

Rheum franzenbachii Munt. is a plant that belongs to the genus Rheum in the family Polygonaceae. Its root and rhizome are used as medicines which are bitter and cold. It has the effect of purgation, purging heat and loosening the bowels and can be used in the treatment of constipation, jaundice, carbuncle sore, furunculosis, burn and scald, and so on. In addition, it is antibacterial, antiphlogistic, convergence, hemostatic, etc.In this paper, using Rheum franzenbachii callus as materials,the study on different types and different concentrations of plant growth regulator in the proliferation effect of suspension culture,and its effective constituent-rhaponticin was detected by high performance liquid chromatography(HPLC),DPPH free radical method was used to explore antioxidant activity in the alcohol extract of different samples. which lay a foundation for the further development and utilization in Rheum franzenbachii. The main experimental results are as follows:1. Cell suspension culture of Rheum franzenbachii,the proliferation of the effect is not obvious when with single growth regulator,one single auxin proliferation effect is better,compared with the control cell growth,almost no browning phenomenon,MS+2,4-D2.0mg/L is the best medium single factor,cell culture content fresh weight was9.43g,increased3.14times,dry weight was0.60g,increased2.00times.2. Different concentrations6-BA(0.5,1.0,2.0,4.0mg/L) and2,4-D (0.5,1.0,2.0,4.0mg/L) combination, the cells of MS+6-BA0.5mg/L+2,4-D2.0mg/L presented straw yellow, Proliferation the best effect,the fresh weight quantity reached12.05g, and increased4.02times; the dry weight quantity reached0.73g, and increased2.43times.3. Different concentration of TDZ and NAA combination, not suitable for cell suspension culture of Rheum franzenbachii, proliferation of cells produce part Browning and dispersion is bad.4. Callus were incubated on the fluid medium MS+6-BA2.0mg/L+NAA1.0mg/L, the growth circles both were39days, and the growth-curve of fresh weight and dry weight looked like "S", the growth circle could be divided three parts:delay phase,index phase, and steady phase.5. The contents of rhaponticin were detected by HPLC. Rhaponticin content of3kinds of cell culture content in9choosen callus (medium types were MS+TDZ0.5mg/L+NAA0.2mg/L,MS+6-BA2.0mg/L+NAA1.0mg/L,MS+6-BA1.0mg/L+2,4-D2.0mg/L) were higher than wild plant(4.36mg/g), in which the highest(medium type was MS+6-BA1.0mg/L+2,4-D2.0mg/L and rhaponticin content was17.64mg/g) was4.05times greater than wild plant's root. 6. There were significant differences among the extracts of nine samples on DPPH radical scavenging ability, the cells'alcohol extract of MS+6-BA1.0mg/L+2,4-D2.0mg/L had the lowest IC50(15.644μg/ml). The alcohol extract of root of wild Rheum franzenbachii's IC50is2.477μg/ml, lower than Vitamin C (2.502μg/ml).