原位转化获得转基因大豆方法的建立
Establishment of in Planta Transformation of Soybean

DOI: 10.12677/BR.2014.35025, PP. 195-199

Keywords: 大豆，原位转化法，BnTR1，根癌农杆菌
Glycine max (L.) Merr., In Planta, BnTR1, Agrobacterium tumefaciens

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Abstract:

大豆是公认的较难转化的植物之一，农杆菌转化大豆是大豆遗传转化中最常用的方法，但是目前的转化效率仍很低。本实验采用原位转化法将具有耐热功能的基因(BnTR1)导入大豆，具体方法是大豆种子萌发、伤口制造、农杆菌侵染、诱导新芽分化，最后用100 mg/L的PPT(草丁膦)筛选鉴定抗性苗。通过该方法成功将pc3301-121-BnTR1质粒转化大豆中，经PCR检测，共获得6棵T0代阳性植株，转化率为4.96%。
Soybean [Glycine max (L.) Merr.] is recognized as one of the plants which is very difficult to be transformed. Agrobacterium-mediated transformation method is the most commonly used method in soybean transformation. However, it results in a relatively lower transformation rate. In this study, in planta transformation was used to introduce thermal resistance gene (BnTR1) into soybean. The experiment process was through seed germination, wounds making, Agrobacterium infection and new buds induced afterwards. Finally 100 mg/L PPT (glufosinate) was used for screening resistant plantlets. Through this method, we successfully introduced plasmid pc3301- 121-BnTR1 into soybean and got resistant plantlets. With approve of PCR detection, six T0 trans-genic plants harboring BnTR1 were obtained with a transformation rate of 4.96%.

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