农杆菌介导的PSAG12-IPT基因转化柳枝稷研究

为延长生物能源作物柳枝稷(Panicum virgatum)的叶片功能期,以柳枝稷‘西稷1号’品系的成熟胚诱导的愈伤组织为受体,采用农杆菌介导法转化叶片衰老延缓基因P_SAG12-IPT.结果表明:经过农杆菌转化,共培养,抗性愈伤组织筛选、再生和生根培养,在农杆菌介导处理的8500块愈伤组织中共获得了320株转化植株;经PCR检测分析,转化植株中共有19株植株扩增出标记基因潮霉素B磷酸转移酶基因的目标片段,初步表明P_SAG12-IPT基因已整合到柳枝稷基因组中,转化率为0.223%.对柳枝稷转基因植株的叶片叶绿素含量、净光合速率、胞间CO₂浓度和气孔导度进行测定,表明P_SAG12-IPT基因在部分转基因植株中已经表达.

Agrobacterium-mediated Transformation of PSAG12-IPT Gene in Switchgrass

In order to prolong the leaf functional period of bioenergy crop switchgrass, the leaf senescence gene P_SAG12-IPT was transferred into switchgrass using Agrobacterium-mediated transformation with the embryogenic callus from mature seeds as the materials. The results showed that 320 transgenic plants were obtained from 8500 callus after agrobacterium-mediated transformation, cocultivation, screening resistant callus, plant regeneration and rooting. PCR analyses of transgenic plants indicated that the target fragment of Hygromycin B phosphoric acid transferase gene was amplified from 19 plants. It preliminary showed that the P_SAG12-IPT gene was integrated into the genome of switchgrass, and the transformation efficiency was 0.223%. Further research on the leaf chlorophyll content, net photosynthetic rate, intercellular CO₂ concentration and stomatal conductance of transgenic plants showed that P_SAG12-IPT gene was expressed in some of the transgenic plants.