荔枝體細(xì)胞胚胎發(fā)生及種質(zhì)超低溫保存研究.pdf

1、湖南農(nóng)業(yè)大學(xué)碩士學(xué)位論文荔枝體細(xì)胞胚胎發(fā)生及種質(zhì)超低溫保存研究姓名：謝玉明申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：果樹學(xué)指導(dǎo)教師：張秋明易干軍2003.6.1StudiesonSomaticEmbryogenesisandGermplasmCryopreservationofLitchiAbstractLitchi(LitchfchinesisSoma)belongstoSapindaceaefamilyandisasubtropicalfruitkn

2、ownforitsdeficiousflavorsAstherearedifficultiesinlitchivitroculture，onlyfewstudiesreportedon$omaticembryogenesisandgermplasmcryopreservationThereforedevelopmentofbiologicaltechniqueislimitednlisexperimentwasconductedonso

3、maticembryogenesisandgermplasmciyopreservationoflitchibyvitrificationTheaimistoestablishprocedureofanthersomatieembryogenesisandsuspensioncellscryopreservation，andtoprovidefoundationforbiologicaltechniqueaswellThemainres

4、earchresultsareaSfo]lows：1AseriesofexperimentshavebeencarriedoutonanthersomaticembryogenesisAntherswereinoculatedonMSmediumcontaining20mg／L2，4一D02mg／LNAAandsupplementedwithsucrose509／LAfter2months，theembryogeniccall／were

5、obtalnedThenitwastransferredtotheMSmediumcontaining05mg／lKT，01mg／LNA氏and05％ACforembryoidsdifferentiationAboutamonthlaterhundredsofembryoidsformedTheseembryoidsweretransferredtotheMSmediumcontaining10mg／LB^10mg／LKT05mg／LN

6、A～500mg／LCH，andsupplementedwithsucrose50∥LCulturedfor23months，theregeneratedD】antlerswereobtalnexLThesystemwasestablishedforsomaticembryogenesisoflitchibyantherculture2PreservationofembryogeniccatliwasstudiedEmbryogenicc

7、alligrewwellonMSmediumcontaining10mg／L2，4D02mg／LNAALongtermpreservationcanbeachievedbycombinedsolidandliquidmediumculturealternativelyNowsomaticembryogenesisispreservedforayearbyaboveculturemannerAtthesametimeembryogenic

8、callihadvigorousgrowthwithembryogeniccharacteristits3EmbryogeulcsuspensioneellswereusedtostudycryopreservationbyvitrificatioILitoptimizedcryopreservationprocedure：take3to5dayageembryogenicsuspensioncellsandpreculturedfor

9、2dayswith04meI／Lglucitol。thenpretreatedwith60％PVS2for15minutesat25℃anddehydratedwithPVS2for15420minutesat0“12Abovesuspensionswereplungediriteliquidnitrogendirectlyandconservedfor24hoursandthenrapidlythawedinawaterbathat4

10、0℃T1levwerewashedtwicewith12meI／LsucrosesolutionTTCanalysisandrecoverygrowthwereemployedRelativesurvivalratereached271％4Ultrastrncturalchangesincryoprescrvedlitchiembryogenicsuspensioncellsbyvitrificationwereassessedusin

11、gtransmissionelectronmicroscopyItwasfousdedthattheplasmolysisbecamesevereduringtheprecultivationdehydrationandcoolingAfewcellswallcellmembraneandnucleusenvelopewerelethallyinjuredduringthecoolingprocess，whileothercellswe