Establishment Of Tribulus Terrestris L.suspension Culture System And Effects Of Different Factors On Flavonoids Content

Tribulus terrestris L.is an annual herbaceous plant,which is also known as Tribulus terrestris,Quite,Tribulus terrestris,Tribulus terrestris,Tribulus bizi.The fruit of Tribulus terrestris is used by dry and ripening fruit that is rich in chemical content.The T.terrestris.is well distributed throughout the country because of the adaptability,and it is more concentrated in the north of the Yangtze River especially the semi-humid semi-arid areas.They are mostly born in sandy land,barren hills and mountainous areas.Autumn harvest fruit is concocted medicine after dried out and impurities.Now the Tribulus terrestris pollution-free standardized planting technology research has been carried out in many places due to the production.Tribulus terrestris can be used for sandy land restoration because of ability of roots to spread to a wide range of areas and playing a wind prevention and sand fixation function,with strong drought resistance.In this study,the stem segments of the T.terrestris.were selected as the explants on the basis of the existing aseptic seedlings.The best explants were selected for the selection of the callus of T.terrestris,and the concentration of each nutrient was adjusted.Stem segments were selected and the best solid callus medium was selected for the sterile T.terrestris at 25 ?.MS + 6-BA 0.1 mg/L + 2,4-D 0.5 mg/L+ ZT 0.5 mg/L.Get a bright color,shiny,loose texture,good growth of the cell line.By selecting good cells to keep the spare,the liquid suspension Was cultured.On the basis of the above,the liquid suspension culture was prepared.The first factor was adjusted and compared with the single factor of the concentration of the hormone in the suspension culture of the T.terrestris,and the growth amount was selected as the observation index.The optimum concentrations of 6-BA,2,4-D and ZT were respectively 0.3 mg/L,0.5 mg/L and 0.5 mg/L.Under these conditions,the three hormones were combined to determine the best medium of suspension for callus cells: MS + 6-BA 0.1 mg/L + 2,4-D 0.3 mg/L + ZT 0.5 mg/L.After determined the main medium,the callus of T.terrestris was able to grow normally under the appropriate environmental conditions and factors.In this study,the growth factor of the T.terrestris was used as the observation index,and made the inoculation amount,medium,rotation speed,pH value,concentration,light cycle optimization.The contents of flavonol in the suspension cells were 1.91%,1.54%,0.56% and 0.61% respectively under the optimum conditions for the suspension culture of T.terrestris as 25 ~ 30 g/L,the best medium as MS,the rotation speed as 120 r/min,the pH as 6.5,origin of carbon as 30 mg/L,and 1/2 light culture for 30 days to obtaine the optimum suspension cells.In order to further improve the content of flavonol of cells,the addition of inducible phenylalanine,tyrosine and cinnamic acid was carried out.The optimal concentration was MS + 6-BA 0.1 mg/L+ 2,4-D 0.3mg/L + ZT 0.5 mg/L + 9.39 mg/L phenylalanine +9.56 mg/L tyrosine +10.94 mg/L cinnamic acid,the chemical composition and flavonol content increased by 0.78%.In this case,the liquid suspension culture system of T.terrestris was not only established but increased the content of flavonol.