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1、上海交通大學(xué)碩士學(xué)位論文北美喬松胚性組織成熟胚形成潛力差異的生化基礎(chǔ)研究姓名：范建芝申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：園林植物與觀賞園藝指導(dǎo)教師：申曉輝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
