Research On Rapid Propagation Technology And Extraction Optimization Of Medicinal Ingredients Of Dioscorea Zingiberensis C.H.Wright

Dioscorea zingiberensis C.H.Wright is a perennial twining herb of Dioscoreaceae,Dioscorea L,it is endemic to our country.Due to the fact that the wild resources of Dioscorea zingiberensis C.H.Wright have been harvested for many years,the wild resources of Dioscorea zingiberensis C.H.Wright have been greatly destroyed.The traditional propagation method has a long cycle and limited resources.It is urgent to find a method of rapid propagation.Dioscorea zingiberensis C.H.Wright contains a variety of medicinal ingredients such as flavonoids,polysaccharides,and dioscin.With the expansion of the use of these medicinal ingredients,it is of important economic value to explore effective extraction methods.In this experiment,the buds segment of Dioscorea zingiberensis C.H.Wright were used as explants to induce differentiation into buds,and the axillary buds were proliferated and cultured,and then the roots were proliferated separately to establish a tissue culture and rapid propagation system of Dioscorea zingiberensis C.H.Wright;And the response surface method was used to design the extraction of flavonoids,polysaccharides and dioscin from Dioscorea zingiberensis C.H.Wright,to investigate the influence of various factors on its extraction,to optimize the extraction process,and to provide a theoretical basis and technical reference for more effective utilization of Dioscorea zingiberensis C.H.Wright resources,better improve the economic value of Dioscorea zingiberensis C.H.Wright.1.The buds segment was the most suitable explant for induction of Dioscorea zingiberensis C.H.Wright.Disinfected with 75%alcohol for 30 s,rinsed with sterile water for 3 times,disinfected with 0.2%Hg Cl₂for 2 minutes,and rinsed with sterile water for 5 times before inserting,the induction rate of the medium was the highest,reaching 96.7%.2.In the experiment,different media types and hormone concentrations had significant effects on the induction of axillary buds.Using MS medium,the induction effect was best.When 6-BA 2 mg·L^-1and 2,4-D 1 mg·L^-1were added,the induction rate can reached 92.8%,the plant height was 4 cm,the fresh weight was 1.5 g,and the dry weight was 160.9 mg.3.In the process of multiplication of Dioscorea zingiberensis C.H.Wright,when the hormone combination was 6-BA 2.5 mg·L^-1and NAA 0.75 mg·L^-1,the multiplication effect was the best,the multiplication coefficient was 8.2,the plant height was 4.8 cm,the fresh weight was 0.54 g,and the dry weight was 56.2 mg.4.When the roots of Dioscorea zingiberensis C.H.Wright were cultured individually,the effect was better when 1/2MS+6-BA0.5 mg·L^-1+NAA1mg·L^-1+sucrose30 g·L^-1were selected,the extension of the root reached 2 cm,the fresh weight was 0.86 g,and the dry weight was 84.9 mg.The use of liquid medium was conducive to the proliferation of roots.The extension of roots reached 3.1 cm,the fresh weight was 0.93 g,and the dry weight was 94.2 mg.5.The optimized extraction parameters of Dioscorea zingiberensis C.H.Wright flavonoids were ultrasonic time 48 minutes,liquid-to-material ratio 39 m L·g^-1,and ethanol concentration 84%.Under these conditions,the actual extraction rate can reached 22.57%,and the total reducing power of the extracted flavonoids was 86.86%,the scavenging rates of DPPH,O₂^·-and·OH were respectively 93%,37%and 97%,and it had strong antioxidant capacity.6.The best technological parameters for the extraction of polysaccharides from Dioscorea zingiberensis C.H.Wright were ultrasonic time 31 minutes,material-to-liquid ratio 52 m L·g^-1,and ultrasonic temperature 39°C.According to this condition,three parallel tests were carried out,and the polysaccharide yield can reached17.92%,and its antioxidant capacity was strong,the total reducing power was 84%,and its scavenging rates for DPPH,O₂^·-and·OH were 33%,87.6%and 66%,respectively.7.This experiment used the combination of enzymatic hydrolysis and ultrasonic extraction,and the response surface optimization design method was used to investigate the effect of extraction conditions on the extraction rate of dioscin,and finally obtain the optimal extraction conditions for enzymatic hydrolysis time 1 h,enzymatic hydrolysis temperature 40℃and material the liquid ratio was 20.5 m L·g^-1,and the yield of dioscin can reached 5.33%under these conditions.In addition,its total reducing power was 34.3%,and its scavenging rates for DPPH,O₂^·-and·OH were 16.7%,16.4%and 18.4%,respectively,and it had certain antioxidant capacity.The technological conditions proposed in this study can be used for the extraction of flavonoids,polysaccharides and dioscin from Dioscorea zingiberensis C.H.Wright,the optimized results are highly reliable and can be used for industrial scale production.