愈伤诱导和继代条件优化.docx

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1、nhighefficientca1.1.usp111.iferatonprotoco1.andIimonoidsandDictnmnineaccumu1.ationofDictamnusdasycarpusTiircz.,ava1.uab1.emedicina1.p1.antinChinaCa1.1.usinduction,pro1.iferationfrommature1.eafcxp1.antsand1.imonoidsaccu1.ationofDictanmtsdasycarpusAbMn1.CiDkvwwiwvdasycrptisTurczIsapcrennia1.va1.uedher

2、bintbcRacracfami1.y.Inthisstudy,ahighCfneSenIca1.1.usinductionandpr。IifCnHionrocoifromyoungstemcxp1.antsofDicu(Sanyaiqwha.beenCMiibIishedand成分含量.Ca1.1.uswasinitM1.FrImtbcynungMcmcxpbntsiubiitcdonMrsedxignficamUi1.Yervn1.m1.e.11ehighestUidixIiDnfvqey(M)O%)WkWIknVedcu1.1.swnsobscn,cdonMSmediumsupp1.em

3、entedwith1.0mg.,1.X4-D拿伤TDZinCombeiwiionwithNAAWaSverysuitab1.eforCaHsro1.itctii(ioncomparedioTDZinCOmtMnaiMMiwith2.4-0TOZincombtnatowithIBA.Freshanddrywe砌ofca1.1.uscu1.turedonMSmediumsupp1.ementedwith0.5mgMSediunMippaenicii1.edwith0.Smg,1.Kincmbta(invi1.h2.0ng,1.NAAWXsimi1.;irothatofwi1.dphnKandtbc

4、contentof)MVum.rraindk1.it*m1.im川th。”Rhan11.obtainedw*whheandfrbb1.e.Keywords:DidMCUSdawErPux、Ca1.hwir1.c(ion.Ca1.1.1.npro1.iferation,1.imnnMhAbbreviationsBAP6-RcnzyhiniifKipunecNAA-rup1.hancJE1.iCvidWkIMMf1.1.j：UtEIi1.idB4n1.o(u1.fwfifrac)xi.CmZwyrmXq*sr.Gmhtoyg1.IMeyhw:3)IO2J1.xtCTVMdn.*eftMoi1.fk

5、DkuruUytfpuRwdaeMitry.IO72D2。沁hSkpboxyMreiMacidMSMuniihigcandSkggftttIMXf1.；”KimMKimHSnIaGGmAnW.C1.”SI:AiSHMnrttfnry11MHeofDkurnud*yiftTu/.fo)1.N1JIiefme3ickfJfkjIinixnArrwakc.JJhuf4u11uc*nwQIrhvwyjri.*im*nip-MK5ang.Xuan.1.J.XuY.MAZang.J.S.2002.Cyktoxc103Qgdandren*no944WMenOSC，搂r11ntherootberkEaMAsi

6、a.hercHbarkofdiePIanaisca1.1.edCitrtexJicfamniintheQieePAIrnIaCrPneEwhichKIKCdinChinese1.rddition1.mcikri11cforIbcIrcatmcntofjaundice,cough,r1.untism攸iBm1.SOmCindicde才小H1.有效成分娥及其畲二，*tt1.jBMI*201VSihAi4c(5dca(ukoof1.iauM.Jk1.amiiH4)auwcMXIGaijndknc1.ogica1.1.yJtctivccocnponcntshavebeeni加Mifiedfromthe

7、11xbarkofI),datyrarpusschas1.imonoids,furoquino1.inca1.hitoids.f1.avonoids,coumarins,sesquiterpenes,scquitcncg1.ycoMde.andptc11uhcg1.ycmitk47收避AmDIttUw1.n.1.imonoid,diet;IJ1.Im1.-ObiHr1.nXm:XHM1.Iiuxine1.kKM!areIhCnnMdetiMmWEedrepreenuive11.iMM*：-N06PEtrgJIuMCMUTi1”o1.k*AtdbyhifiiperkotiMM风向CtgM(UwI

8、yfc(i*X、CaMtiwatp,u*1.*isCvCteGDktSrnIiEfuMR.&ParXJ.H.e.Extic1.ku1.o(R*tBMkofIXt1.imiMdMbV4*.pdf3XC11wpMrt*0Q1.f*mwinixtiatfyof3Eu1.ni1.fmmDktXnWqh11rppdf拧置苦素的活性1.imonin.oneofthemajorCompooetsfoundinthep1.anu1.,RuiaceaeandMcIiACCiieOimiHcx.h;tbwncop1.ayvariniwbkngca1.m1.cincancer,inammatiix).a111.ob

9、cshyinmanydifferentCd1.卜PCSand1.is、UaJnd3PmySSqiunifkumMUrfroicvii、Cacii、kicagainsg1.uanwc-iducxincurocoxiciymprimaryCUkUrCgofra1.xti1.CdkIh1.1.*ttIMMUt*201S-D*ruryohuue*ftf411ran1.inub*nBM90tnfty.IM1fcytriyvrrm*mdnby.pj(p-ManyMudiexhavebeenrePwtedth；uobiicunnewasfoundtnjrvat1.yenhanceIheCy1.otoxici

10、cyOfmicr1.II1.档活性HuxiiieIkMieexhibit%avanv1.yofin1.vcMnvnX!rties.inc1.udingIbean1.inIIainina1.orvbowe1.disease.DCUmPm1.CC1.ive,andimcctic1.a1.actiitienc.MEIMMtt二X1.1.UmdUah1.”Jv1.b.De1.mcn1.EfYeUt4I4tie1.kjcrMmWf3)I6-Wcw1.(djpert(rnBWWKttDictamninc.anatura1.p1.antpduci.hasbeendcmonstrutod(ohavesigni

11、fkuntCfTOe1.Sonan1.i-p1.akJFKreWin.uxau1.ar-rtfhinj:.ancs.Kft1.ftCo.”.t11*wd%r*hti%v0MTihrkofixmgvp1.1.nrKucwc)gInconc1.usionIjmnnin.dictii11ninc.nbcuno11ca111.frAxinc1.1.ne.themajoractiveConyiKHe，ofD.dasaw.hasthepotentia1.Hignihcam1.yInmPtf6(1.wna1.ura1.pppuhtiDntouicreaM：.OverIhtfyear*.I1.ienota1.

12、bark,who1.ePbnIOfD.daxyrarpwtiscontinuous1.ydean*CtivdyIUrVCQedfromthewi1.dandoverexp1.oitedbyhuman,thepopu1.ationofwi1.dresourceisincrcusing1.ydev1.inedofdisnppcarodfrmitsIKMuni1.1.uibi(.Ine而改HHiOnofthecreasingdenwik1.thereisaneedcodeve1.opnewapproachesfwMis1.ainab1.cdeve1.opmentaxiuti1.i4i1.innAM1

13、tt1.11eruchCirC1.nnNIrUre、,invitrocu1.turetechniquesC(H1.k1.1.utinn1.heyPmbkrn5,bee;IU糅i1.mayrpd1.yirrc%ctheproductionofbiomassuxJtheaccumu1.u(ionofeconomica1.1.yimportantScCQndUrymetabo1.itesinaCiMnParIi1.iVe1.ys1.ut1.imexucnpaudIuc加柳.4dt9F究内21Xf1.t义1.OUIkus1.edge,1.1.u:efcen(ca1.1.usptu1.iferatMM

14、iwMtxro1.ax1.hiuae(ivecus1.i(u(esof1.dasycarjmhasxbeen)erformedunti1.ven1.,11*erefure.IheobjectiveofIbecurrentresearchwastoikvdancffkicn1iIcrationofdasarptisandfurtherana1.ysisOf1.bCactiveMy1.oChCmiCakintheca1.1.uscu1.ture.Materia1.sandmethodsExpUnhandca1.1.usinductionTheshootsofDdasveatpuswereCH1.U

15、rCdforXMaysontheMSmediumcontaining0.5mg-1.BAPaxi0.05mg/1.NAA.inadditionco301.sucroseand8g1.agabasedonprevious1.yCMabIishedregra1.iunprotoco1.unpub1.ished).YouiigstemM!C1.iu(abc1.1.5cn1.ength)Irxxii4mmXwhichthesbxttiphj1.beenremovedwereCXCiMrdandusedascxphn1.x.Ixxca1.1.usinduction,stemMXbonswereincub

16、atedinMSso1.idmediumWPPMnKInCdWiIhChIKrcmcocmrutionofNAA0.0.0.5.1.0.2.0.3.0.4.0or5Ang1.)JBA.eachwith60cxpknperIroi1.mcn1.Cu1.tureHereincubateda(26*2X2IHMkr30T(In1.InISTPEI)photon11uxhr16hetcyofca1.1.usIfkfuctiunwasex,a1.ua1.ed31.er45dofcu1.ture,rmin*cI1i.Ca1.1.usprohfera(hmKorIbeca1.1.u111.ttruinUXp

17、tfnmm1.XIket1.ec1.ofvariuutcovmu1.icnofBAP(0.0.0.25.0.5.10.2.0or4.0mg1.)orTDZ(0.0.0.25.0.5.1.0.2.0or4.0mg.1.)BKT(0.0.0.25.05.1.0.2.0or4.0ng,(1.)inCOmbiMiOnwithNAAOJmg;1.)or(0.5mg.,1.jor2.4-D0.5mg1.)WaScrfondmitheMSmedium.fttttt*811mc11WrCBefUIbrmcutojreve+y“drredefreeIiumoffiIrU1.tudMIkTCdMIhCMntIif

18、cvwt1.Ki.wuU1.cdKUkwab1.11ff11v11.mimm6川.JnMhrwvidccmirMEj询次Nndt1.MawEachca1.1.usHsric(grownfor45d)of02g(FW)0.04BDW)wainitia1.incubatedonIr1.enmeyerf1.ask,withIOrep1.icasperIrCinmE.Ca1.1.usIrrCSHc*hiandHryWdhWaAca1.mtod45dafcrcu1.turerespective1.y.Growthindex(Gi)wascaku1.atedaxIbedescnbedby*nw;i5fu)

19、fWcightruCa1.1.iweredriedm60Cfbr48h.Ca1.1.usgrowthcurve1.ttK!Qt2:A11nrutikttr1.!frnhuv1.X,in;ixix*vhOnIUMfTWfcxhfcw4rtrminnxgrue11Uca1.1.usincutxrtedPertr1.en11cyef1.ask,a1.ca1.1.uswasco1.1.ectedandEgU1.nrdfrc*handdryweightCVCry5d.respective1.y.Quantitativeana1.)EofUmonoids方法:IFIHtheqIKSi1.ita1.ivea

20、na1.yiufbiuc1.ivephyUK-IieinicaIs,1.0gofeuthponderedca1.1.usax1.Wik1.P1.anI、nate11a1.wasIhMmixed1.h25n1.nct1.uuu1.ande.1.ractedIwuIinwsU1.Xkrhea1.-tv11ux一attMm511jA:RPHPtXMuM*门好成中3H活住成少的aMf11r1.h,andthencoo1.inganddix*o1.vcwithmc1.h;inn1.co25m1.TheCnmhinCdextw1.sucrcfikcrcdthrough0.22mmcmbuncbeforeH

21、P1.Cana1.ysis1.1.1.方法2Hjjtheqx1.i(a1.iveanu1.yt%biue(ivephyKK1.ieinicaIs.1.0gofeachpoderedca1.1.usaidfttt1.M(*f1.1.trvitofIiHiitvi.dkmnc.ciMconcmuJtrnt1.1.mcWfRHHP1.C川UMI/d戌中3H1.tt成分的介0一物WtVI.Fdrwikiphnmateria1.s*af1.11tmixedwith25In1.ctha11n1.water(75:25,vv)andcxtnctc1.IWCtimCAbybeat-ref1.uxingfor2

22、h.TbcCX1.nK1.iOaso1.utionswerecombinedundfi1.tratedthrough0.22mmcnbrane.AfterrecoveringC1.hano1.thercudua1.aqueousWaSCXUaCIedotimesbyIOn1.CH2CI2.1.he111.1.CH2C12extrac1.vaunxu1.trdtodrynessuungraryevajHra1.wa1.amaximumtemperatureof45U1.1.JI巾.ThCc1.31.sWCrCdkokedeihIom1.methano1.bcfreHP1.Cana1.ysis.S

23、anpkana1.ysiswasper1.ataf1.owru1.eof1.0m1.min.Thedetectionwave1.engthandOO1.unW(cmpcra(urcwasu236nmand35,心PeV1.i、C1.y.A1.!retcrence%UiixkirdswerePUrd1.amx!fromna1.iuu1.Uis1.i1.u1.etI1.iecon(m1.ofp!uracvutica1.ax1.bio1.ogica1.pnx1.uc1.TheconsOfxxmsamp1.e*weredeterminedby1.heexterna1.5tankrcimethod.1.

24、1.CYpCrimCnIAIdatawereXMtiMicaIIyana1.yzedbyonewayana1.ysisofvariance(NOV)IWinEthetestP0.05).andWCrCP1.VMnMCdbyMCragC+andardemu.Reu1.t%4nddhcusimCa1.1.%inductionYoungstemCXPhnMwereincubatedonMSMMidmediumUipp1.emcntcdWithdiI1.cren1.CanCC1.UnHion(V1.1.rying1.eve1.s)ofNAA.IBAand2.4-DIbrcu1.1.usinductio

25、n.写法!1.eexp1.ainsbecamesu1.1.ena1.ter7*Me11u1.ca1.1.yfrwnIheGIMmaci11NoftheCipIanKafter14ixMSsupp1.ementa1.WithNor2.4-D.(2Ht写法2ComptandIighbgrccn-Co1.onx1.ca1.1.ideve1.opedfromthecutmarginsofICIIfcxp)anisafterI4dofcukurc.whi1.eye1.1.owishandfriab1.eCaniwereinducedafter30d1.ightgreenvtriab1.eiriwhite

26、GiHUXwerenbuinedfromyoungQCmexphntsafter30dCUhUrcdix1.subcu1.curciCVefy30d.1.BiE9形态特征)含伤形在Ca1.1.usIib1.aiixx1.nthisIIHX1.iU1.nWaSwhi1.e.f1.andfriab1.eThePCTcCntfrCqUencyoca1.1.usi11i1.ia1.infmmthecxp1.ant%waspresentedinIdbkI.TheresponsewasbetteronMSMJPpknIEtOdwithBAPandNAA.Tbcca1.1.usinductionra1.er

27、eachedM100%onMSWPP1.anCMedMihXXX.muchhigherhanocherUCMmCm工However,thefrequencydecreasedtc20%nnMSmediumWPP1.CmC1.KCdwithX(和Mt比，找下降点).SomctxxJyreportedthat(be某个SHR是有效的,和本wasc111.cien(forca1.1.usinduction,andthisisVCrysimi1.artoourresu1.t帝成鱼伉酎同的变化IiKreasein(1.wCQnNnUIHigGfNAAfrCMn2Io4ng,1.and0.5ng.1.K1

28、miMSmediumde1.ayedni1.ia(nof2.4-DWaSmoresuitab1.eforCaHi1.Sinducin.Who*theOmEntnHicoofX4Duuahigh。52.0R1.IiBombinMionwith0,R1.BAP.theCMIMinductsr,i1.crC4dwhifrfI1.unM1.crIrciiHtcnUTWcI)W1.ieiiIhtfMvc1.a*of2,4-duu%inrcudH*4.(r;1.-.ihtfca1.1.usi11dctkmgcdcvrvaedhup1.y1.85-1.8.0Tab1.eIEtTectofp1.antgro

29、wthrcu1.oonca1.1.uiuktnfhninvjiwyx三*aemcxp1.antsofD.dancatpMEMur7mMSafk，3)d1.dDCMkOttWAQ(MM)5%*5-4nHcnhWtjDiyCiJ1.MS3MSI0.55UO.I2O7O.OI392IXI0.13560.0172洗黄球310O.3570.0179*淡黄嫁43。KU(M460OOIW淡黄绘54。U1.1.O.520.0ISf淡黄65X1I1.1.1.0.15280.0199I，求黄70.5250XI990.01海81.080.13280.0138.荷92.0ni1.0.2478O.O26I决黄绿IO3.

30、0I1.1.1.0.6450.0160J法业娘I1.4.0I1.1.1.0.15910.015714n125.0III)0.14950.0148r),臼130.514IX1.152163.0174.0IX5共计壮U1.4t三*C6X.)a.1U族外体，次IS=卯，M2ft.生.”kcdg1,.gnMfi1.GrtvWuhiU*,Y)v1.bu,B1.1.ifcratkmIntheca1.1.us1.ifaioncxpc11nts,theCiYeC1.ofvariousConeniraikxuofTDZ.BAPundKTinCtHnbina1.iunwith0.5n1.NAAUrsSma1.2.

31、4-Dscreenedak,ewasPerh1.mKXJOnMSmedium.不好的WhenTDZincombinationwithNAA.IhcircucOffPhanddryweightofCaHI1.Swa,nxnobb1.cundmaximumfreshanddryweightwos3.7Igand0.3IRrespective1.y.TheCfTcC1.ofTDZinxnbnwith2.4-DwasdtMincandmaximumfreshuddyvig1.incrcaa1.to89311I0.45g,rcparedwithTDZinCOn1.hinationwith、AA.和上的脩

32、对比：XHtMJKMSmediumsu1.cmcnt!ifera1.ioncnwith2.4DorIAA.Fre4Weigh1.anddryWCixh1.ofca1.1.uvcu1.turedonVISmediumMipp1.cmcntcdwith0.5mg,1.TDZincomhi11iinixwith0.5mg,1.2.4-DWCrC1.3J5gand0.61.g.increased263nxJ15.0times,respective1.y.InJMk第二个我.可以讨的Theeffectofca1.1.uxpro1.iferationwasveryXigninCInIonMSsuppkme

33、niedWiihdi伍m1.BAI,inCOTnbinution0.5mg1.NAA.Freshweightanddryweightofcu1.1.usWCIV10.46gand0.65g.increase20.9and15.5times,cu1.turedonMSmediumsupp1.ementedWi1.h2.0mg1.BAPinCombinaoonwith05ng1.NAA.Thepro1.iferationeffectofBPinCCmhinMionwith2.4-Dw;ixSmihirto6-BPincombinationwithNAA.HOWCVCr.themuximumfres

34、handdryweightsdecreasedto4.84gand037gonMSmediaCOiMaining6-BAPincombiwionwithIAA.respccivc1.y.优正方Ofa1.1.mediatried.MSEediInnContiining2mgI-I2,4Dand0.5mjjMKinwasthebc1.forca1.Iuvinitiationantirc1.itc11tion.Ca1.1.usingresponsedecreaseddrastica1.1.yto58%rsuhinginpooca1.1.usformationonIhiSmedium.Ontheotherhand.NAAMbeconceniraikx)ofIOmg1.1.and0.5mg1.1.KimreMdIedear1.yinitii(ion(4-6day*amiverygox1.pro1.iferation.ErTaC1.ofBP.KT.TDZincm1.wna(iwith(IMMKkWmtYmca1.1.uspr1ivm(onUCD.(Arrvoirprrr刖次数HAhKnnW1.ITUZ1.nu1.i2.4-DHitf1.1.OfNAAgXsfh1.1.aGr,wCdIiKMi1.1.K05I1.0而透泳愎J2X1堂底床混43.0Witiftes40