细胞悬浮体系的建立及成分测定.docx

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1、的鹿悬浮体系的建立及成分测定Estab1.ishmentofCe1.1.suspensioncu1.turesofDictainnusdasycarpusfortheproductionofbioactivecompounds(Dictaminine)Obacunone(M46M),fraxinc1.1.ou(MM),IimoniiM柠苦素)Dktamnine(白鲜WD目的,1 .建立ce1.1.suspensionprotoco1.:2 .确定影响产量的因素3 .确定最优配方内容,1 .P1.antmateria1.andestab1.ishmentofce1.1.suspensionCan

2、US的培养,*期已建立的体系(引用发表文章)ce1.1.suspensioncu1.ture(用BUE方)t2.0gFW=S50m1.250m1.=J过漉=反复SUbCUI1.urC2 .PGRsconcentrationandcombination激素选择:2.4-D.NAA.IBA:0.1.0.5,1.0.2.0.5.0mg1.激素组合:2.0mg1.2,4-D+KTorBAP:0.1,().5,1.0,2.()mg,3 .cu1.turecondition(1) Mediastrength!1/2、1/3、1/4、1.0MS(2) Inocu1.umdensity:2.5,5,1(),1

3、5,20,25gFW(3) Carbonsource(concentration):I,2.3,4.5,6.7%(4) PhotoperiodtDark.16-1.ightf24-1.ight(5) Initia1.1.1:4.0.5.().5.5,6.0,6.5,7.0,8.0,9.0,10.()(6) KH2PO4concentration:(7) Mg2+Concentration!370mg1.(8) NH4+NO3:总氏(1:1)20-100:比率:60:0=():604 .Growthcurve(kinetics)1 -8weeks5 .Dctcrminationofce1.1.b

4、iomassThrough0.45umstain1.essstc1.1.sieve(sigma).FWandDW(60度)6 .ExtractionandHP1.Cana1.ysisDiscussioni1. 类对生长和含量的影2.4书对生长量的影对成分的影响I2. 光曝对生长量和成分含量的影响:3. 初始PH对生长和成分含的影响;4. M2+对生长的影响及其机制(号始5荤甲妣径)5. NH4+浓度对生长量和含量的影*及其机M(气基荤甲媾径)Estab1.ishmentofCe1.1.suspensioncu1.turesofDic1.amnusdasycarpusortheproductio

5、nobioactivecompounds(Dictaminine)Estab1.ishmentofCe1.1.suspensioncu1.turesofDictamnusdasycarpusandbiosynthesisofactivecompounds(Dictaininiiic)AbstractKeywords:Introduction写作思路:药用植物介绍(是个要的药用植物,作为传统中药;功效;活性)主要成分是*植物本身含量低(heusua1.1.yvery1.ow)Recent1.y,重要成分Ahasattractedinterestedduetoitsstrong*.的功效,活性,治

6、疗的疾病在植物上veryminorProdUCtion,TherefOrc,a1.ternativeexp1.orationof*isimportant,.、的前人研究进展hasbenreportedin*本文研究内容Materia1.sandmethodsP1.antmateria1.andestab1.ishmentoce1.1.suspensionInf1.uenceof1,(;RsonbiomassandtheproductionofbioactivecompoundsOptimizationofcu1.tureconditions(1) Mediastrength:1/2、1/3、1

7、/4、1.0MS(2) Inocu1.umdensityt2.5,5,10.15,20,25g-FW(3) CarbonSoUrCe(ConCentnitiOn):I,2.3,4.5,6.7%(4) Photoperiod:Dark.16-1.ight.24-1.ight(5) Initia1.pH:4.0.5.0,5.5.6.0.6.5,7.0,8.0,9.0,10.0(6) Mg2+Concentration:370mg1.(7) NH4+Concentration:1650mg,1.Growthcurve(kinetics)Determinationofce1.1.biomassQuan

8、titativeana1.ysisofbioactivecompounds制用自己文章ExtracispreparationandquantitativeHPI.Cana1.ysesofactivecompoundswereperfomwd.accordingtotheproceduresdescribedPreVioUs1.y1引自己发表文章)Statistica1.(Data)ana1.ysis1.1.experimenta1.datawerestatistica1.1.yana1.yzedbyonewayana1.ysisofvariance(ANOVA)usingtheprotecte

9、d1.east-signit1.cant-dierenee(1.SD)es(P0.05),andwerepresentedbyaveragestandarderror(SE).Resu1.tsanddiscussion生长素Conccntration(mg1.1.FnhWdRhtgWeight(g(?activecompOimdN1.mWXCe1.statusObacuimiwFraxine1.1.oneIJnMMiInDicCimnine2.4-D0.0O.I0.20.030.20.03IO.2O.O3IO.2O.O3IO2O.O3IO.2O.O3Pa1.cye1.1.ow,f11gi1.e

10、OJ1.02.0Darkye1.1.ow,cxnpact5.0NAA0.00.1OJ1.O2.05.0IHA0.00.10、1.02.05.0分裂素,固定2,4-D为2.0m1.)CancentnitiDr,Weight1.jContentsofactivecmpounds(mggDW)Ce1.1.MAtmOKacum*Fni*d1.onc1.imoninPk1.amnincKiN).00.110.20.03IO.2(BIO.21O.O3ION0.0310.20.031320.03PaJeyei1.ou,.(ruti1.e0.51.02.0Di1.rtjr1.1.ow.CEnPMM5.()BA

11、P0.00.1OJ1.02.05.0Mediastrength序号MSStrengthFreshWeightContentsofacthecnpounds(nggDWCe1.!%tiA1.uOhuvunoneFmxine1.kmc1.im(mtcn1.ofactivecmpounds11gDVV)Oi1.statusObavunoncFraxinv1.1.one1.1.nnContentsofacthrCOfnPO1.mdS1.m血DVVCe1.1.statusOhiacunoneFnixinc1.1.one1.imoninOidamnineI02I10.20.031O.2O.O310.20.

12、03IO.2O.O310.20.031020.03Pa1.cye1.1.ow,fragi1.e3243546576Photoperiod序号PhiMopcriOdFrwhWeight(g1.DryWeightWCtxofctivcUHnIMMInxr1.1.ow.fragi1.e3241.ighInitia1.pH序号Initia1.pHFreshVvi1.1.DryWeightWContent%ouc1.iecompnd%1.DW)CtUstatusObjcuiKineFraxiiwUGnc1.iinuninDkr1.ainnineI4.025.010.20.0310.20.0310.20.

13、03IOJ0.031O.2O.O310.20.03Pakye1.1.ow,fragi1.e35.546.057.068.079.0Mg2+Concentration序号Mg2*Concentra1.io11g1.JFEhWeiut()DryWeightContcntxofactiveCimPeHIrKh1.m隙DWCe1.1.sUninDictainnincI0.0020.2030.37IO.2O.O31O.2O.O3IO.2O.O3IO.2O.O310.20.03O.2O,O3Pa1.er1.1.ow.f11gik40.6050.8061.00NH4CIConcentration序号NHCo

14、nccntni1.ion(uM)Fre%hWeight(f)DryWght(Contcn1.MofactivecpoundHncgDWCe1.1.MatuxObacunoneFraxhwUoneIJnMn1.nDk1.amnineICK(MS)20.8010.20.0310.20.031O.2O.O3IO.2O.O310.20.03IO.2O,O3Pa1.cXv1.1.ow.f11gik320.6(1.65g42.0052.5063.00注1加入KNO3(摩尔号)保持NO窝子浓度不交,用NHQ提升N川浓度应外加入KNo3,1650+80.0434(mIttQ*101.10氧化帔)=2.OX4s

15、1.,此时保证了稽根平行.MS中正富NH4NO3(NH4,)浓度:1650+X0.0434(硝M)=2O,611M克与摩尔换算,克数子*子量(分子量)=物腐的量(mo1.敷)比如麻,气的物质的=8/16=0.5m”KO-TIO1.Ift1.NH4C1.5.49NH4NO3分子量8(MM54NH4+Concentration序号NH4CIconcentratin1.g1.)Fr%hWciht(三)DryWeight(KContcntxofxc1.ivcCImPotIndXImMC1.)WCe1.1.sumsObacunDncFruwUunvIJ11Mnin1.)k1.aninrI0.0020.8

16、010.2003IO.2O,O3I0.20.0310.20.0310.20.0310.20.03Pa1.eyCU0%fragi1.e31.6542.0052.5063.00注:保持、。3离子浓度不受,用、川C1.提升、H/浓度Grothcurve(kinetics)序号Cu1.tureT1.mc1.wk1.FreshDryWefcht(g)Contentsofactivecmpounds(mggDWCe1.1.%U1.uxObacunoncFraXiIK1.1.owIJmoninDktamnincII223310.20.031O.2O.O310.20.03IO.2O.O310.20.03IO2O.O3Pa1.cye1.1.ow,f11gi1.c445566778K

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