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以新海30和新海16的胚性愈伤组织为转化受体,用农杆菌介导法将苏云金芽孢杆菌杀虫晶体蛋白(Bt)基因导入新海30和新海16。结果表明,在相同的转化条件下,新海30遗传转化效率大于新海16。培养基中无机盐浓度为1/2MS,凝固剂gelrite浓度为2.0 g/L时,能够提高胚状体萌发和成苗率,降低畸形苗频率。通过T1代PCR及T2代RT-PCR检测证实Bt基因已整合到了海岛棉基因组中,获得转基因植株。
The embryogenic callus of Xinhai 30 and Xinhai 16 were used as the transformation receptor, and the Bacillus thuringiensis insecticidal crystal protein (Bt) gene was introduced into Xinhai 30 and Xinhai 16 by Agrobacterium-mediated transformation. The results showed that the transformation efficiency of Xinhai30 was higher than that of Xinhai16 under the same transformation conditions. When the concentration of inorganic salt in culture medium is 1 / 2MS and gelrite concentration is 2.0 g / L, the germination and seedling rate of embryoid body can be increased and the frequency of deformity seedling can be reduced. The T1 generation of PCR and T2 generation of RT-PCR test confirmed that the Bt gene has been integrated into the island cotton genome and obtained transgenic plants.