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Application1:DefinitionofSHComparisonofSHwithSexualcrossingPossibleFusionResultsofTwoGeneticallyDifferentProtoplastsTerminology-1Terminology-2Technicaldevelopmentalhistory2:protoplastfusionFusionmethods1.PEG-mediatedfusionCharacteristics:1)Meritsinclude:lowinput,highfrequencyofhetero-karyons,nospecieslimit;2)Defectsinclude:tediousfusionprocedure,toxicitytocellsWorkingmechanism:PEG分子具有轻微的负极性，可与具有正极性基团的水、蛋白质和碳水化合物等形成H键，从而在原生质体之间形成分子桥，从而使原生质体发生粘连进而促进原生质体融合；PEG能增加类脂膜的流动性，也使原生质体的核、细胞器发生融合成为可能Fusionsol:CaCl22H2O8-10mmolKH2PO40.7mmolMannitol0.5-1.0molpH5.6Inducingsol:Fusionsol+PEG20-45％Dilutingsol:solA(g/100ml)pH6.0solB(g/100ml)pH10.5Glucose7.21Glycin0.375CaCl22H2O0.79NaOH0.169P12．Electrofusion,ElectricallyinducedfusionThreeadvantagescomparedwithPEGfusionNotoxicitytoprotoplastsHigherfusionfrequencySimplermanipulationElectro-fusioninstrumentinNKL,HZAUTheprocessofelectricallyinducedfusionFusionparametersAlternatecurrentvoltage(交流电压)TimedurationofAC(交变电场处理时间)Directcurrentvoltage(直流高频电压)Pulsewidth(脉冲宽度)No.ofPulses(脉冲次数)FusionprocessFactorsaffectingPPfusion
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