| Objective: To study the effect of different mineral elements in MS medium on the content of secondary metabolites in the callus of Pueraria mirifica.To study the effects of different levels of exogenous rare earth elements,methyl jasmonate and acetylsalicylic acid on the secondary metabolites and physiological and biochemical in the callus of Pueraria mirifica.To further optimize the culture conditions of Pueraria mirifica and provide new technical means and data reference for the rapid propagation system of Pueraria mirifica.Methods:(1)According to the HPLC method established in the previous experiment,to study the effect of Cu,Mg,Zn,Mn and Ca in MS medium on the secondary metabolites in the callus of Pueraria mirifica.by Orthogonal test method.(2)To analyze the effects of different levels of rare earth elements,lanthanum nitrate,cerium nitrate,cerium ammonium nitrate,and methyl jasmonate and acetylsalicylic acid on the secondary metabolites of puerarin,genistein,daidzin and total flavonoids in the callus of Pueraria mirifica.(3)According to physiological experiment technique,to detect the effects of different factors on Physiological and Biochemical Indexes of SOD,MDA,POD and CAT.Results:(1)The range analysis results of puerarin content were 0.012,0.016,0.013,0.012,0.08,and the range analysis results of genistein content were 0.116,0.071,0.162,0.195,0.199;the range analysis results of daidzin content were 0.023,0.016,0.013,0.012,0.008,the range analysis results of total flavonoids were 0.139,0.065,0.174,0.182,0.180.The main factors affecting the content of puerarin in the callus of Pueraria mirifica were Ca> Zn> Cu> Mg> Mn,and the best condition of the medium was A3B1C1D3E1.The main factors of daidzin were Ca> Mn> Zn> Cu>Mg,and the best condition of the medium was A1B3C3D4E4.The main factors affecting the content of genistein in the callus of Pueraria mirifica were Cu> Mg>Zn> Mn> Ca,and the best condition conditions was A1B2C3D1E2.(2)The optimal cerium nitrate concentrations for puerarin,genistein,daidzin and total flavonoids were100?mol/L?0.1?mol/L?100?mol/L?100?mol/L,and the contents were increased by159%,132%,120% and 112% compared with the blank control group.The optimal Lanthanum nitrate concentrations for puerarin,genistein,daidzin and total flavonoids were 0.1?mol/L?10?mol/L?10?mol/L?10?mol/L,and the contents were increased by 63%?787%?695% and 624% compared with the blank control group.The optimal Ceric ammonium nitrate concentrations for puerarin,genistein,daidzin and total flavonoids was 10?mol/L,and the contents were increased by 272%?89%?67% and81% compared with the blank control group.(3)The optimal cerium nitrate concentrations for SOD?POD?CAT?MDA were 100?mol/L?100?mol/L?10?mol/L?100?mol/L,and the enzymatic activity were increased by 136% ? 707% ? 182%compared with the blank control group,and MDA content decreased by 70%.The optimal Lanthanum nitrate concentrations for SOD?POD?CAT?MDA was 10?mol/L,and the enzymatic activity were increased by 177%?1062%?201% compared with the blank control group.,and MDA content decreased by 78%.The optimal Ceric ammonium nitrate concentrations for SOD?POD?CAT?MDA was 10?mol/L,and the enzymatic activity were increased by 169%?332%?251% compared with the blank control group,and MDA content decreased by 55%.(4)The optimal Methyl jasmonate concentrations for puerarin,genistein,daidzin and total flavonoids was 100?mol/L,and the contents were increased by 281%?909%?375% and 418% compared with the blank control group.The optimal Acetylsalicylic acid concentrations for puerarin,genistein,daidzin and total flavonoids were 1?mol/L ? 0.1?mol/L ? 100?mol/L ?100?mol/L,and the contents were increased by 260%?114%?368% and 305%compared with the blank control group.(5)The optimal Methyl jasmonateconcentrations for SOD?POD?CAT were 100?mol/L?100?mol/L?10?mol/L,and the enzymatic activity were increased by 135%?329%?99% compared with the blank control group,and Methyl jasmonate will increased MDA content.The optimal Acetylsalicylic acid concentrations for SOD?POD?CAT?MDA were 100?mol/L?10?mol/L?10?mol/L?0.1?mol/L,and the enzymatic activity were increased by75% ? 315% ? 249% compared with the blank control group.,and MDA content decreased by 29%.Conclusion:(1)The different types and contents of mineral elements in MS medium had some influence on the secondary metabolites in the callus of Pueraria mirifica.(2)The rare earth elements,methyl jasmonate and acetylsalicylic acid could promote the accumulation of secondary metabolites and physiological and biochemical in the callus of Pueraria mirifica.(3)The optimal culture conditions and the main influencing factors of puerarin,genistein,daidzin and total flavonoids were different.Therefore,we should choose different culture conditions to obtain the maximum product content when culturing and extracting the single secondary metabolites. |
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