PMI基因为选择标记植物表达载体的构建及在雪柑转基因中的应用

以PMI基因替换植物表达载体pCAMBIA1301中的hpt基因以及pBI121中的GUS基因，构建了以PMI基因为选择标记基因的植物表达载体pCAMBIA1301PMI和pBIPMI，并导入根癌农杆菌EHA105中。研究了两种表达载体对雪柑上胚轴的转化，在培养基附加25 g•L-1甘露糖和5 g•L-1蔗糖为碳源的选择压下，pCAMBIA1301PMI的转化率为27.7％，pBIPMI的转化率为12.7％，对再生植株进行了氯酚红检测和PCR检测，建立了以PMI/甘露糖为选择系统的雪柑转基因体系。

The construction of plant expression vectors with PMI gene as selection marker and utilization in transformation of ‘Xuegan’

The plant expression vectors pCAMBIA1301PMI and pBIPMI were constructed by substituting Escherichia coli phosphomannose- isomerase (PMI) gene for hpt gene of pCAMBIA1301 and GUS gene of pBI121. Epicocyl explants of ‘Xuegan’ sweet orange were inoculated with EHA105- pCAMBIA1301PMI and EHA105- pBIPMI and subsequently selected on medium supplemented with a combination of 25 g•L-1 mannose and 5 g•L-1 sucrose as a carbon source. The transformation efficiency rate was 27.7% when transformed by pCAMBIA1301PMI and 12.7% by pBIPMI .Genetic transformation was confirmed by Chlorophenol Red assay and PCR. A new method for obtaining transgenic ‘Xuegan’ plants was developed using PMI/mannose selection system.