【摘要】 用根癌农杆菌(Agrobacteriumtumefaciens)介导法将水稻巯基蛋白酶抑制剂基因(OCI)导入甘薯(Ipomoeabatatas)品种栗子香,获得了转基因植株。所用菌株为根癌农杆菌LBA4404,其携带的pBinh质粒上含有新霉素磷酸转移酶Ⅱ(NPTⅡ)基因和OCI基因。将继代培养3d后的栗子香胚性悬浮细胞与LBA4404(OD600nm=0.5)共培养4d。将共培养后的胚性悬浮细胞首先在含有2mg/L2,4-D和300mg/LCarb(carbencillin)、但不含有Kan(kanamycin)的MS液体培养基中培养5d,然后在含有2mg/L2,4-D、50mg/LKan和300mg/LCarb的MS液体培养基中进行选择培养。选择培养4周后,将直径约1mm的抗Kan细胞团转移到添加2mg/L2,4-D、50mg/LKan和300mg/LCarb的MS固体培养基上,共转移200个细胞团,诱导得到了8个胚性愈伤组织。将这些抗Kan胚性愈伤组织转移到添加1mg/LABA、50mg/LKan和100mg/LCarb的MS固体培养基上,通过体细胞胚胎发生途径获得了13株再生植株。PCR和PCR-Southern检测结果表明,获得的13株再生植株中7株是转基因植株。更多还原

【Abstract】 Regeneration of sweetpotato (Ipomoea batatas cv. Lizixiang) transgeni c plants with oryzacystatin-I (OCI)gene was achieved using an Agrobacterium tume faciens mediated method. A. tumefaciens strain LBA4404 harboring a binary vecto r pBinh with NPTⅡand OCI genes was used. Lizixiang embryogenic suspension cultu res of 3 d after subculture were cocultivated with LBA4404 (OD600nm=0.5) for 4 d . After cocultivation, the infected suspension cultures were first cultured for 5 d in MS medium with 2 mg/L 2,4-D and 300 mg/L Carb (carbencillin) but without Kan (kanamycin) and then transferred into MS medium supplemented with 2 mg/L 2,4-D, 50 mg/L Kan and 300 mg/L Carb for the selection culture. Four weeks after se lection, 200 Kan-resistant cell aggregates about 1 mm in size from the embryogen ic suspension cultures were transferred to MS solid medium supplemented with 2 m g/L 2,4-D, 50 mg/L Kan and 300 mg/L Carb and 8 embryogenic calluses were obtaine d. After transferring to MS medium supplemented with 1 mg/L ABA, 50 mg/L Kan and 100 mg/L Carb, these embryogenic calluses formed 13 plantlets via somatic embry ogenesis. PCR and PCR-Southern analysis indicated that 7 of 13 plantlets were tr ansgenic plants.更多还原