利用pmi标记基因筛选培育转EaDREB2B基因甘蔗

利用已构建的植物表达载体prd29a-dreb-hyg，通过酶切连接到含有磷酸甘露糖异构酶基因（pmi）的表达质粒pZMLR14上，构建植物表达载体pDREB-PMI。利用基因枪轰击转化甘蔗愈伤，经过甘露糖筛选，共获51株抗性苗，转化再生频率为4.25%。对转基因植株进行分子检测，结果表明有8株为阳性转基因无性系。氯酚红试验表明标记磷酸甘露糖异构酶基因在转基因株系中均有表达。对转基因T₁代甘蔗植株进行分子检测，结果表明EaDREB2B基因在转基因甘蔗无性系T₁代中稳定遗传。该结果为进一步研究EaDREB2B基因在甘蔗抗旱方面的作用奠定了基础。

Transformation of EaDREB2B Gene into Sugarcane Using pmi Gene as Selectable Marker

The plant expression vector of pDREB-PMI was constructed successfully with expression vector of prd29a-dreb-hyg and pZMLR14 by restriction-ligase reaction. Sugarcane callus were transformed with pDREB-PMI expression vector by bombardment. After mannose selection, 51 regenerated plants were obtained with the transformation rate of 4.25%. All of these 51 seedlings were detected with molecular detection, and the 8 seedlings were positive transgenic sugarcane. Chlorophenol red assay of transgenic sugarcane leaf showed that the pmi gene was expressed in transgenic plants. T₁-generation transgenic sugarcane were detected by PCR and RT-PCR, and the exogenous gene EaDREB2B was stably inherited. The results laid the foundation of further research on the effect of EaDREB2B gene in improving sugarcane resistance to drought stress.