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1、植物学通报2006,23(3):275~280ChineseBulletinofBotany.组织培养简讯.结球白菜下胚轴原生质体培养及其体细胞胚植株再生1*12刘凡赵泓秦帆(1国家蔬菜工程技术研究中心北京100089)(2四川省农业科学院园艺研究所成都610066)摘要结球白菜(Brassicacampestrisssp.pekinensis)的原生质体培养由于基因型依赖性强,细胞易褐化,愈伤组织的芽诱导率低等而难于再生植株。本实验以结球白菜的下胚轴原生质体为试材,研究了影响其细胞分裂及愈伤组织形成的因素,探索了经过体细胞胚发生途径获得再生植株
2、的技术。结果表明,试材的基因型及培养基组成影响细胞分裂及褐化;KM8P是结球白菜原生质体培养更适宜的培养基,能显著减轻细胞的褐化;液体培养基中一定浓度的活性炭能在一定程度上减轻细胞褐化进程,并有利于星状细胞团的形成;基因型Asko中,愈伤组织形成体细胞胚的结构,其发生的频率约为5%,该类体细胞胚能全部顺利地发育成完整植株。本技术具有再生植株形成容易、频率较高且通过体细胞胚发生途径等优点。关键词结球白菜,体细胞胚,原生质体培养,植株再生TheProtoplastCultureofBrassicacampestrisssp.pekinensisand
3、PlantRegenerationviaSomaticEmbryogenesis1*12FanLiu,HongZhao,FanQin(1NationalEngineeringResearchCenterforVegetables,Beijing100089)(2InstituteofHorticulture,SichuanAcademyofAgriculturalScience,Chengdu610066)AbstractSomefactorsblockingtheprotoplastregenerationofBrassicacampestri
4、sssp.pekinensisincludegenotype-dependentcellbrowningandlowfrequencyofshootinduction.Screeningforagenotypethatiseasytocultureandestablishinganexperimentalsystemforefficientshootinduc-tionwillgreatlybenefitthegeneticinnovationofB.campestrisssp.pekinensisviasomatichybridization.
5、HypocotylprotoplastsofseveralvarietiesofB.campestrisssp.pekinensiswerecultured,andfactorsinfluencingcelldivisionandbrowningwerestudied.Celldivisionandbrown-ingwereinfluencedheavilybygenotypesofmaterialsandtheculturemediaused.KM8Pmediumeffectivelyreducedthebrowningtendencyofth
6、ecellsandwasagoodmediumforprotoplastcultureofB.campestrisssp.pekinensis.Activatedcharcoalhadaspecificfunctionofreducingcellbrowningandpromotingthestar-shapedmicrocalliformation.Thesomaticembryoswere-1observedoncalliofthegenotypeAskoinshoot-inducingmediumcontaining3mg.LAgNO3,w
7、ithafrequencyofabout5%.Intactplantsweredevelopedeasilyfromtheembryos.Thisproce-durehasadvantagesineasilyregeneratingplantswithrelativelyhighfrequencyviaasomaticembryogenesispathway.KeywordsBrassicacampestrisssp.pekinensis,somaticembryogenesis,protoplastculture,plantregenerati
8、on收稿日期:2005-12-22;接受日期:2006-03-09基金项目:北京市自然科学基金(5050001)和北京市科技发展计划项目
