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利用农杆菌介导法,以东北地区种植的9个大豆基因型和国外品种Williams82的子叶节为外植体,将抗虫基因cry1Iem转入栽培大豆品种中。共转化外植体1 459个,获得84棵再生植株。采用除草剂叶片涂抹法、PCR及Bar蛋白试纸条检测法对得到大豆再生植株进行鉴定,转cry1Iem基因大豆T_0阳性植株为61株。对部分T_1转基因植株的遗传分析表明,外源基因能够稳定遗传到下一代。通过对部分T_1阳性转基因植株进行Southern blot分析,证明目的基因片段均已整合到受体大豆的基因组DNA中,单拷贝率为22%左右。对获得的部分T_2材料进行了室内抗大豆食心虫鉴定,有2份转基因材料抗性较对照显著提高。
Nine Agrobacterium-derived soybean cultivars and Williams82 cotyledonary node planted in Northeast China were used as explants by Agrobacterium-mediated transformation. The cry1Iem gene was transferred into cultivated soybean varieties. A total of 1 459 explants were transformed into 84 regenerated plants. The soybean regenerated plants were identified by the method of herbicide leaf smear, PCR and Bar protein strip test, and the 61 transgenic plants with cry1Iem gene soybean T_0 were identified. Genetic analysis of some T_1 transgenic plants showed that foreign genes can be stably inherited to the next generation. Southern blot analysis of some T_1 transgenic plants showed that the target gene fragment had been integrated into the genomic DNA of the recipient soybean with a single copy ratio of about 22%. Some of the T_2 materials obtained were identified as indoor anti-soybean Moth beetles, and the resistance of two transgenic materials was significantly higher than that of the control.