不结球白菜OguCMS下胚轴原生质体培养再生植株

Plantlet Regeneration from Hypototyl Protoplast Culture of OguCMS,Non heading Chinese Cabbage

取不结球白菜OguCMS 5d苗龄的下胚轴 ,在 2 0mg/mL纤维素酶 (OnozukaR 10 ) ,10mg/mL果胶酶 (MecerozymeR 10 )及CaCl2 ·2H2 O 10mmol/L、KH2 PO4 0 .7mmol/L、甘露醇 0 .5mol/L的酶液中游离 36h ,50 0r/min下离心收集下胚轴原生质体。原生质体在KM8P1附加 2 ,4 D 0 .5mg/L、 6 BA 0 .2 5mg/L、NAA 1.0mg/L的培养基上培养 2 1d形成小愈伤组织 ,经MS附加 2 ,4 D 1.0mg/L增殖愈伤组织 ,在MS附加 6 BA 10 .0mg/L和NAA 0 .3mg/L培养基上进行芽分化培养 ,并在无任何激素的MS培养基上生根 ,14d后即可获得完整的再生植株。

Hypocotyl from 5 day old seedling was digested in enzyme solution containing cellulase Onozuka R 10 20mg/mL,pectinase Mecerozyme R 10 10mg/mL,CaCl 2·2H 2O 10mmol/L,KH 2PO 4 0.7mmol/L and mannitol 0.5 mol/L for 36h,then centrifuged at 500r/min.The protoplast were collected and cultured on KM8P 1 medium with 2,4 D 0.5mg/L,6 BA 0.25mg/L and NAA 1.0mg/L.Microculli formed after 21 days culture,then multiplicated on MS medium with 2,4 D 1.0mg/L.On MS medium containing 6 BA 10.0mg/L and NAA 0.3mg/L,they were initiated shoots,and then transferred to a hormone free medium to develop roots.Plantlets from protoplast culture were regenerated in 14 days.