杂种番木瓜亲本材料的体胚发生及基因导入

Somatic Embryogenesis and Gene Transformation of Hybrid Papaya Parental Materials

不同品种番木瓜体胚再生条件的比较表明:亲本材料在 NAA1.0mg/L+KT1.0 mg/L 的改良 MS 培养基上可诱导出愈伤组织,在 KT0.5mg/L+NAA0.25mg/L+活性炭0.3%的改良 MS 培养基上诱导体胚,经诱导的愈伤组织需在 NAA0.5mg/L+KT0.25mg/L 的培养基上继代一次才可诱导出体胚。利用此再生系统,以胚性愈伤组织作为转化材料,与农杆菌 LBA4404(pRPCY/pAL4404)共培养后,在选择培养基上诱导出体胚出芽,生根培养基(IBA 2.0mg/L)上生根。从小植株中提取总 DNA,PCR 检测结果初步显示,抗性筛选的11个株系中有7个有 PRV CP 基因插入。

Abundant calli of parental materials of hybrid papaya were induced on modified MS medium(MMs)supplemented with NAA 1.0 mg/L,KT 1.0 mg/L,and subcultured on MMS supplemented with NAA 0.5 mg/L,KT 0.25 mg/L.The somatic embryos can be obtained on MMS supplemented with KT 0.5 mg/L,NAA 0.25 mg/L,activated charcoal 0.3% .Embryogenic calli were transformed with A.tumefaciens LBA 4404(containing pRPCY/ pAL 4404).Buds were formed on the selection medium and rooted on root medium(including IBA 2.0 mg/L).Total DNA was extracted from 11 lines for PCR and the results infered that the PRV CP gene may be integrated into the genome of 7 lines.