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1、上海交通大学硕士学位论文北美乔松胚性组织成熟胚形成潜力差异的生化基础研究姓名：范建芝申请学位级别：硕士专业：园林植物与观赏园艺指导教师：申晓辉20070120STUDIESONTHEBIOCHEMICALBASISOFMATUREEMBRYOSFORMATIONPOTENTIALDIFFERENCEOFPINUSSTROBUSEMBRYONICTISSUEABSTRACTDuringtheexperimentusingtwocelllinesofPinusstrobus(No.01andNo.02)t
2、oinducematuresomaticembryos,itwasfoundthatonlytheNo.01celllinehadthecapacitytoformmatureembryos.So,weattemptedtopredictearlierET’spotentialabilitytoformthematuresomaticembryosthroughcomparingdifferencesinET’smorphologyandstructureduringproliferationcult
3、ure.ByanalyzingETgrowth,sucrosemetabolismandproteinexpressionduringtheproliferationstage,weaimedatprobingintothemorphology,cellularandmolecularbasisinthepotentialofinducingmatureembryos.Theresultswereasthefollowings:(1)Thetwocelllineshadnosignificantmor
4、phologicaldifferencesinproliferationstages,whilehaddifferenceinthestructure:No.1celllinehadatypeofstructurenamedESM4whichwaslackofinNo.2cellline.(2)FromtheRGRcurvesofETs,itwasfoundthattheRGRofcelllineNo.2washigherthanNo.1celllinethroughoutthewholeprolif
5、erationculturestageandreachedtomaximumduringtheforthandfifthday,whichhadadelaycomparedtoNo.1cellline.TheRGRofNo.1celllinereachedthtomaximumonthethirddayanditwasverylowonthe15dayandwasthclosedto0onthe20day.(3)Therewassomecorrelationamonghexosecontents,th
6、erateofhexose/sucrose,activitiesofInvenzymesandRGR.(4)ThetotalsolubleproteincontentsweremeasuredbyBradford’smethod.Generallyspeaking,No.1celllinewassignificantlyhigherthanNo.02intotalsolubleproteincontent.II(5)WestudiedproteinexpressionofETintwocellline
7、sduringproliferationphasebySDS-PAGE.Theresultsindicatedthat,inthebothofcelllines,thereexistedsomeproteinsthatthemolecularweightswereabout88.8KD,64.2KD,53.8KD,36.5KD,33.9KDand18.6KD.However,thequantityof18.6KDinNo.1celllinewasmorethanthatinNo.2cellline,t
8、hequantityof36.5KDinNo.1celllinewaslessthanthatinNo.2cellline.Inaddition,thereweresomespecialproteinsinNo.2celllinesuchasproteinswith71.6KDand24.0KD.ThecellmicrostructureofPinusstrobusfromproliferationtomatureembryoinductionwasfi
