Establishment Of Silybum Marianum Cell Suspension Culture System And Effects Of Different Inducers On Silybin Content

Silybum rnarianum(L) Daertn. is an 1-2 annual herb of Silybum, Cynareae Less, Asteraceae. It was originally planted in Europe, Africa, and the Mediterranean region. The herb likes a cool and dry climate and is not strict in growth environment. With its adaptability, it can grow on barren hills, or areas of poor fertilizer. Generally, its leaves are mined in spring and collected in summer. With its cold, bitter taste, the plant is a good hepatoprotective plant and is widely used in clinical treatment of liver disease. In addition to its hepatoprotective effects, it also shows a variety of other pharmacological effects, such as the protection of cerebral nervous system, cardiac muscle cell, the protection of the skin, anti-inflammatory and good anti tumor effect.With the rising of its clinical application demand, the increasing scarcity of wild resources degradation and cultivar, Silybum marianum is in short supply. Plant tissue culture could alleviate the shortage situation, but there is no real effective bioreactor culture of plant tissue. In this experiment, through the cultivation of S. marianum leaf explants, calli were obtained and mass cultured. On this basis, suspension culture condition is optimized, the final optimum conditions of suspension culture are determined as follows: MS liquid medium, light culture 12h/d, pH=7, 110r/min, 20 d, culture hormone ratio for 6-BA 1.5 mg/L + NAA 1 mg/L + ZT 1.5 mg/L, cell suspension structure is loose, the color yellow, medium clean and clear, divided into small granular cell.In establishing the elicitor culture system, suspension is added to determine silymarin suspension cell growth and silybin. The silybin in silymarin solid original plant, callus, and liquid elicitors without addition of suspension in culture were 0.046%, 0.349%, and 0.501% respectively. Compared with them, addition of sodium nitroprusside is not only conducive to cell suspension of weight gain, but also effective to silymarin accumulation. When sodium nitroprusside concentration was 1 mmol/L, the amount of growth of Silybum marianum suspension cells reached maximum, 12.24 g, and silybin content reached 0.686%. Followed by chitosan, with the concentration of 3 mmol/L, silymarin suspension cell growth is 11.73 g, silybin content of 0.616%. Salicylic acid produces a negative regulation on silymarin suspension cells, with maximum growth of Silybum marianum suspension cells as 5.32 g, silybin content only 0.146%. The content of silybin in suspension culture is 1.43 times of that in the solid callus, silybin content in sodium nitroprusside group is 1.36 times that of culture liquid without elicitors, and 1.96 times the solid callus. Thus, suspension culture does good for the enrichment of Silybum marianum cell weight and secondary metabolites.This study investigates the influence of different factors on the effect of silymarin suspension culture system, determines and optimizes silymarin suspension culture. On this basis, different elicitors are added, secondary metabolite in silymarin suspension culture cells are accumulated increasedly, and a theoretical basis for the silymarin suspension culture is provided.