细胞悬浮体系的建立及成分测定.docx
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.suspensionCanUS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(),15,20,25gFW(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.biomassThrough0.45umstain1.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.amnusdasycarpusortheproductionobioactivecompounds(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、1/4、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.biomassQuantitativeana1.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)usingtheprotected1.east-signit1.cant-dierenee(1.SD)es(P<0.05),andwerepresentedbyaverage±standarderror(SE).Resu1.tsanddiscussion生长素Conccntration(mg1.1.Fn»hWdRht<R>gWeight(g>(?<»ntcnUt>activecompOimdN1.mWXCe1.statusObacuimiwFraxine1.1.oneIJnMMiInDicCimnine2.4-D0.0O.I0.2±0.030.2±0.03IO.2±O.O3IO.2±O.O3IO2±O.O3IO.2±O.O3Pa1.cye1.1.ow,f11gi1.eOJ1.02.0Darkye1.1.ow,c<xnpact5.0NAA0.00.1OJ1.O2.05.0IHA0.00.10、1.02.05.0分裂素,固定2,4-D为2.0m1.)Cancentniti<mImWIjFreshWdgh1.>Dr,Weight1.j>Contentsofactivec<>mpounds(mggDW)Ce1.1.MAtmOKacum>*Fni*>d1.onc1.imoninPk1.amnincKiN<).00.110.2±0.03IO.2(BIO.21O.O3ION±0.0310.2±0.03132±0.03PaJeyei1.ou,.(ruti1.e0.51.02.0Di1.rtjr1.1.ow.CEnPMM5.()BAP0.00.1OJ1.02.05.0Mediastrength序号MSStrengthFreshWeight<g1.DryWeight1.j>Contentsofacthec<>n>pounds(nggDW>Ce1.!%tiA1.u×OhuvunoneFmxine1.kmc1.im<minIMcIamnincIIM21.1U.2±O.O3o.2(B10.2±0.051(I2±O.O310.20.0310210.03Pa1.cye1.1.ow,fragi1.e31/241.0Inocu1.umdensity序号Inocu1.umdensity(R14FEbWdghtIR)Dr,Wcht(>(<mtcn1.×ofactivec<mpounds11gDVV)Oi1.statusObavunoncFraxinv1.1.one1.1.n<minDictaninineI2.5025.(WIO.2±O.O3IO.2±O.O31O.2±O.O310.2±0.0310.2±0.0310.2±0.8Pa1.cx!1.ow.f11gik310.0415.0520.0625.0Carbonconcentration序号C.'arbonConcentratk>n<%)FreshWeight(u)DryWcht>ContentsofacthrCOfnPO1.mdS1.m血DVV>Ce1.1.statusOhiacunoneFnixinc1.1.one1.imoninOidamnineI02I10.2±0.031O.2±O.O310.2±0.03IO.2±O.O310.2±0.03102±0.03Pa1.cye1.1.ow,fragi1.e3243546576Photoperiod序号PhiMopcriOdFrwhWeight(g1.DryWeightWCtxofctivcUHnIMMIn<171.)V)Ce1.1.statusObacunuiwKraxinc1.1.une1.imuninDictaxnnineIDiUk261.ight10.2±0.030.2±0.03IO2±0.0310.2±0.0310.2±0.0310.2±0.03Pa1.c>xr1.1.ow.fragi1.e3241.ighInitia1.pH序号Initia1.pHFreshVvi1.1.DryWeightWContent%ouc1.iecomp<>nd%1.DW)CtUstatusObjcuiKineFraxiiwUGnc1.iinuninDkr1.ainnineI4.025.010.2±0.0310.2±0.0310.2±0.03IOJ±0.031O.2±O.O310.2±0.03Pakye1.1.ow,fragi1.e35.546.057.068.079.0Mg2+Concentration序号Mg2*Concentra1.io11g1.JFEhWeiut()DryWeight>ContcntxofactiveCimPeHIrKh1.m隙DW>Ce1.1.sU<uObacunoncFraxiiwUoncU11M>ninDictainnincI0.0020.2030.37IO.2±O.O31O.2±O.O3IO.2±O.O3IO.2±O.O310.2±0.03O.2±O,O3Pa1.er1.1.ow.f11gik40.6050.8061.00NH4CIConcentration序号NHConccntni1.ion(uM)Fre%hWeight(f)DryWght(>Contcn1.Mofactivec<11>poundHncgDW>Ce1.1.MatuxObacunoneFraxhwUoneIJnM>n1.nDk1.amnineICK(MS)20.8010.2±0.0310.2±0.031O.2±O.O3IO.2±O.O310.2±0.03IO.2±O,O3Pa1.cXv1.1.ow.f11gik320.6(1.65g>42.0052.5063.00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.OX4s1.,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-T>IO1.Ift1.NH4C1.5.49NH4NO3分子量8（MM54NH4+Concentration序号NH4CIconcentrati<>n1.g1.)Fr%hWciht(三)DryWeight(K>Contcntxofxc1.ivcCImPotIndXImMC1.)W>Ce1.1.sumsObacunDncF'ruwUunvIJ11M>nin1.)k1.aninrI0.0020.8010.2±003IO.2±O,O3I0.2±0.0310.2±0.0310.2±0.0310.2±0.03Pa1.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)Contentsofactivec<>mpounds(mggDW>Ce1.1.%U1.uxObacunoncFraXiIK1.1.o<wIJmoninDktamnincII223310.2±0.031O.2±O.O310.2±0.03IO.2±O.O310.2±0.03IO2±O.O3Pa1.cye1.1.ow,f11gi1.c445566778K
