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1、园艺学报2008,35(10):1491-1496ActaHorticulturaeSinica多花蔷薇胚性细胞悬浮系原生质体分离及再生植株3陈颖,梁建丽,陈晓丽,郭艳超,田传卫,赵梁军(中国农业大学农学与生物技术学院,北京100193)摘要:利用多花蔷薇‘无刺3号’假珠芽诱导的胚性愈伤细胞悬浮系分离得到原生质体,并培养获-1-1得再生植株。最佳酶液组成是210%纤维素酶,015%离析酶,510mmol·LMES,015mol·L甘露醇,6-1015%CaCl2·2H2O。采用继代3d的悬浮细胞系,酶解10h,原生质体产量最大(26167×10·g),活力最高(92121%)。采用液
2、体浅层培养,肌醇比蔗糖更有利于纯化后的原生质体分裂和生长。愈伤组织形-1-1-1-1成增殖培养基为1/2MS+110mg·L2,42D+012mg·LEBR+10g·LFicoll+500mg·L谷氨-1-1-1-1酰胺+20mg·L甘氨酸+20mg·L天冬氨酸,在1/2MS+10mg·LTDZ+500mg·L谷氨酰胺-1-1+20mg·L甘氨酸+20mg·L天冬氨酸培养基中分化培养后转入1/2MS培养基上获得完整植株。关键词:多花蔷薇;原生质体;细胞悬浮系;植株再生中图分类号:S685112文献标识码:A文章编号:05132353X(2008)1021491206Protoplas
3、tIsolationandPlantRegenerationfromSomaticEmbryogenicCellSuspensionCulturesinRosamultiflora3CHENYing,LIANGJian2li,CHENXiao2li,GUOYan2chao,TIANChuan2wei,andZHAOLiang2jun(AgronomyandBiotechnologyCollege,ChinaAgriculturalUniversity,Beijing100193,China)Abstract:Protoplastsweresuccessfullyisolatedfro
4、msomaticembryogeniccellsuspensioninducedbypseudobulbils,shootsregenerationwereachieved.Enzymesortandconcentration,treatmenttimeandthecul2turetimeofcellsuspensionshadinfluenceonprotoplastsisolation.Enzymesolutioncontained210%cellu2-1-1lase,015%macerozyme,510mmol·LMES,015mol·Lmannitol,015%CaCl2·2
5、H2O.Optimumenzymetreatmenttimewas10hoursandcellsuspensionssubculturedfor3dayswereadopted.Theyieldand6-1viabilitywere26167×10·gand92121%.Protoplastswereculturedonshallowliquidlayers,whereino2sitolwasbetterthansucroseforpromotingcellsdivisionandgrowth.Calliinducedandproliferationmedium-1-1-1-1was
6、1/2MS+110mg·L2,42D+012mg·LEBR+10g·LFicoll+500mg·Lglutamine+-1-1-120mg·Lglycine+20mg·Lasparticacid.Differentiationmediumwas1/2MS+10mg·LTDZ+-1-1-1500mg·Lglutamine+20mg·Lglycine+20mg·Lasparticacid.Shootsregeneratedon1/2MSmedium.Keywords:Rosamultiflora;protoplast;cellsuspensionculture;shootregenera
7、tion原生质体是很理想的遗传转化受体。蔷薇属原生质体再生困难,成功的报道很少。Matthews等(1991)以蔷薇种间杂种(Rosapersica×Rosaxanthina)的根和光叶蔷薇(R.wichruaiana)的无芽茎段为外植体诱导愈伤组织并悬浮培养,从悬浮系分离原生质体并建立了再生体系;Kim等(2003)以杂种月季(R.hybridaL.‘Sumpath’)合子胚胚性愈伤组织进行悬浮培养,从悬浮系分离原生质体并建立了再生体系。但上述报道的原生
