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1、CharacterizationofproductionofPaclitaxelandrelatedTaxanesinTaxusCuspidataDensiformissuspensionculturesbyLC,LC/MS,andLC/MS/MSCHAPTERTHEREPLANTTISSUECULTUREⅠ.PotentialofPlantcellCultureforTaxaneProductionSeveralalternativesourcesofpaclitaxelhavebeenidentifieda

2、ndarecurrentlythesubjectsofconsiderableinvestigationworldwide.Theseincludethetotalsynthesisandbiosynthesisofpaclitaxel,theagriculturesupplyoftaxoidsfromneedlesofTaxusspecies,hemisynthesis(theattachmentofasidechaintobiogeneticprecursorsofpaclitaxelsuchasbacca

3、tinⅢor10-deacetylbaccatinⅢ),fungusproduction,andtheproductionoftaxoidsbycellandtissueculture.Thisreciewwillconcentrateonlyonthelatterpossibility.Planttissuecultureisoneapproachunderinvestigationtoprovidelargeamountsandastablesupplyofthiscompoundexhibitingant

4、ineoplasticactivity.Aprocesstoproducepaclitaxelorpaclitaxel-likecompoundsincellculturehasalreadybeenparented.Thedevelopmentoffastgrowingcelllinescapableofproducingpaclitaxelwouldnotonlysolvethelimitationsinpaclitaxelsuppliespresentlyneededforclinicaluse,butw

5、ouldalsohelpconservethelargenumberoftreesthatneedtobeharvestedinordertoisolateit.Currently,scientistsandresearchershavebeensuccessfulininitiatingfastplantgrowthbutwithlimitedpaclitaxelproductionorviceversa.Therefore,itistheobjectiveofresearcherstofindamethod

6、thatwillpromotefastplantgrowthandalsoproducealargeamountofpaclitaxelatthesametime.Ⅱ.FactorsInfluencingGrowthPaclitaxelContentA.ChoiceofMediaforGrowthGamborg's(B5)andMurashige&Skoog's(MS)mediaseemtobesuperiorforcallusgrowthcomparedtoWhite's(WP)medium.Themajor

7、differencebetweenthesetwomediaisthattheMSmediumcontains40mMnitrateand20mMammonium,comparedto25mMnitrateand2mMammonium.ManyresearchershaveselectedtheB5mediumovertheMSmediumforallsubsequentstudies,althoughtheyachievesimilarresults.Gamborg'sB5mediawasusedthroug

8、houtourexperimentsforinitiationofcallusculturesandsuspensionculturesduetosuccessfulpublishedresults.Itwassupplementedwith2%sucrose,2g/Lcaseinhydrolysate,2.4mg/Lpicloram,and1.8mg/Lα-naphthaleneac