玉米耐热DNA聚合酶基因表达载体构建与花粉介导转化

Construction of Plant Expression Vector with Taq DNA Polymerase Gene and Gain Heat-tolerant Maize Lines

实验构建热稳定性聚合酶的基因表达双元载体,利用花粉介导法将热稳定性DNA聚合酶基因导入玉米幼胚,获得耐热转基因玉米株系。用热稳定性DNA聚合酶基因构建表达载体;然后利用花粉介导法进行转化,花粉收集自仲恺自交系玉米甜1号和自交系糯2号,将花粉与构建好的基因表达载体混合,利用花粉介导法进行转化;将收集到的T1代种子用PCR凝胶电泳技术和Southern杂交技术进行检测,选出阳性的转化株;在T2代经过田间耐热压力试验结合测定的玉米生理指标,筛选出9个具有良好耐热性玉米株系。

Taq DNA polymerase gene was used to develop new heat-tolerant maize lines, which will potentially add to the economic value and aid in controlling crop losses due to heat stress. First, an expression vector with Taq DNA polymerase gene was established. Then, according to pollen-mediated transformation, pollen from the maize ZhongKai inbred lines, Zhong sweet No. 1 and Zhong glutinous No.2 was collected, which was mixed with the estab- lished expression vector and was applied to the maize silks for transformation. Thirdly, harvested T1 generation seeds were confirmed to be positive transformants by PCR and Southern hybridization. Finally, after screening the T2 generation, by exposure to heat stress conditions in the field combined with physiological indexes, nine trans- formed heat-tolerant maize lines were obtained.