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从东北地区高pH盐碱地上生长良好的羊草中克隆Lc14-3-3基因,以露地菊(Chrysanthemum morifolium)‘火焰’叶片愈伤组织为受体,以农杆菌介导,将Lc14-3-3基因转入‘火焰’基因组中。以载体中的磷酸甘露糖异构酶基因(phosphomannose isomerase,PMI)为筛选标记,通过甘露糖的筛选,结合PCR、Southern杂交检测Lc14-3-3整合到其基因组。通过转基因植株与对照植株的耐盐性对比可见转化Lc14-3-3基因植株具有更高的耐盐能力。磷酸甘露糖异构酶基因/甘露糖筛选系统可有效应用于露地菊遗传转化。
The Lc14-3-3 gene was cloned from Leymus chinensis grown on high pH saline soil in the northeast of China. The leaf callus of Chrysanthemum morifolium was used as the receptor. Agrobacterium-mediated transformation of Lc14-3- 3 gene into the ’Flame’ genome. The phosphomannose isomerase (PMI) in the vector was used as the screening marker, and the Lc14-3-3 was integrated into the genome through the screening of mannose, PCR and Southern hybridization. Compared with the salt tolerance of the transgenic plants and the control plants, it can be seen that the plants transformed with the Lc14-3-3 gene have higher salt tolerance. The phosphomannose isomerase gene / mannose screening system can be effectively applied to genetic transformation of Dendrobium chrysogenum.