Research On Liquid Culture Of Panax Notoginseng Cell And Hairy Root

Panax notoginseng F. H. Chen ?SANQI,ginseng Genus,Araliaceae Family? is the major ingredient of "Yunnan Baiyao". This herb has long been used traditionally to treat cardiovascular and cerebrovascular diseases clinically. However, it takes long time from sowing to final collection, during which many cares are required since the growth is subjected to some factors,such as pathogens and pests,heavy toxic residues and abnormal climate,etc.,which limits the market supply of raw material of Panax notoginseng. Production of Panax notoginseng natural compounds through plant cell engineering, will be useful for solving the shortage of Panax notoginseng resources.Establishment of Panax notoginseng suspension cell culture system includes the following steps, germination of seeds, induction of callus, multiplicational of callus and suspension culture, etc.; In addition, Panax notoginseng hairy roots were gained by interaction between callus and Agrobacterium rhizogenes.Panax notoginseng seeds were found that those seeds treated by 300 mg/L of gibberellic acid for 12 h was peeled and cultured on MS under darkness, 90% of budding rate was achieved at the 45th day of culture. For induction of Panax notoginseng callus,the formula of MS+2,4-D?1.0 mg/L?+6-BA?0.5 mg/L? was proven to be the best, and the culture temperature of 25± 1? was suitable. Moreover, the maximum multiplication of callus was gained when it was supplied with 30 g/L sucrose and 2,4-D?1.0 mg/L?+6-BA?0.2 mg/L?+ KT?0.2 mg/L?.To establish the Panax notoginseng suspension cells culture system, the optimization of culture media was conducted. The formula of MS+2,4-D?1.0 mg/L?+6-BA?0.2 mg/L?+KT?0.2 mg/L?+NAA?0₆ mg/L? supplemented with 30 g/L of sucrose was to be the best?and initial inoculum of 40 g/L could gain the maximum of multiplication of suspension cells and the peak of total saponin content was harvested.When the O.D600 of Agrobacterium rhizogenes bacteria culture got to 0.7, it was used to infect the explant of Panax notoginseng callus for 20 min, after then was transferred to MS and co-cultured for 2 days, and then was transferred to bacterial removing media. The hairy root of Panax notoginseng was tested to be the Ri-transformed roots by TLC of opines. The hairy roots of Panax notoginseng in the liquid media of 1/2 MS grew well,and the content of saponin reached 13.52% at the 30th day of culture, significantly higher than that of the suspension cells and callus of Panax no toginseng .Through research, the liquid culture of Panax notoginseng cell and hairy root with more medicinal ingredients, Panax notoginseng cell and hairy root liquid culture technology provide the basis for industrialized production of Panax notoginseng medicinal product.