Genetic transformation of promising genotypes of winter bread wheat by in planta method

L. Slivka, O. Dubrovna
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Abstract

Aim. Optimization of conditions and genetic transformation of new promising genotypes of winter bread wheat (Triticum aestivum L.) by in planta method. Methods. Agrobacterium-mediated transformation by the in planta method using strain AGL0 and vector construct pBi-OAT. Results. The influence of air temperature, optical density of cells of agrobacterial suspension, inoculation day and composition of inoculation medium on the frequency of obtaining transgenic plants of new promising genotypes of winter wheat was studied. The dependence of the frequency of obtaining transgenic plants on environmental conditions, in particular temperature, has been established. It was found that the temperature regime of 20-22 °C provided the largest number (4.4%) of wheat transformants, and when the temperature is reduced to 16-18 °C there is a decrease in the efficiency of T-DNA transfer into the plant genome and the lowest frequency of transformation is observed. Conclusions. The largest number of transformants was obtained using a inoculation medium without sucrose, the optical density of cells of the agrobacterial suspension of 0.4 op.od. and inoculation on the third day after castration of ears. Keywords: Triticum aestivum L., Agrobacterium-mediated transformation, ornithine-δ-aminotransferase gene.
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冬面包小麦有前途基因型的株内遗传转化
的目标。利用植物法优化冬面包小麦(Triticum aestivum L.)新基因型的条件及遗传转化方法。利用菌株AGL0和载体构建pBi-OAT,农杆菌介导植物内转化。结果。研究了空气温度、农杆菌悬浮液细胞光密度、接种天数和接种培养基组成对获得有前景的新基因型转基因植株频率的影响。获得转基因植物的频率依赖于环境条件,特别是温度,已经确定。结果表明,在20-22℃的温度下,小麦转化体数量最多(4.4%),而当温度降至16-18℃时,T-DNA向植株基因组转移的效率降低,转化频率最低。结论。在不含蔗糖的接种培养基中,农杆菌悬浮液的细胞光密度为0.4 op.od,转化体数量最多。去势后第三天接种。关键词:小麦,农杆菌介导转化,鸟氨酸-δ-转氨酶基因
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