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培育抗草甘膦除草剂棉花品种对棉花生产具有重要意义。利用农杆菌介导法获得了大量抗除草剂草甘膦棉花的再生植株,对其进行阳性检测。通过确定PCR检测目标条带大小、蛋白杂交条带大小分子特征,对65个独立转化事件在DNA水平、蛋白水平进行分析和筛选。结果表明,经PCR检测再生植株DNA得到的阳性率平均值为81.5%,经过蛋白检测后阳性率确定为55.4%,该类阳性植株即为外源基因正常表达的转基因植株。在此基础上,结合田间抗性检测,对其中9个株系进行了不同世代的选择,经过综合鉴定筛选,发现2个抗性优良的转基因株系具有极高的利用价值,其余还需进一步筛选鉴定。
Breeding of glyphosate-resistant herbicide cotton varieties is of great significance to cotton production. A large number of regenerated plants against herbicide glyphosate cotton were obtained by Agrobacterium tumefaciens-mediated method and tested for their positive results. By determining the size of the target band and the size of the hybridized protein bands by PCR, the 65 independent transformation events were analyzed and screened at the DNA level and the protein level. The results showed that the average positive rate of PCR detection of regenerated plant DNA was 81.5%, and the positive rate was 55.4% after the detection of protein. The positive plants were the transgenic plants with normal expression of exogenous gene. On this basis, combining with field resistance testing, 9 strains were selected for different generations. After comprehensive identification and screening, it was found that the two transgenic lines with high resistance value had extremely high utilization value, and the rest needed further Screening and identification.