农杆菌介导phyB基因转化枳壳研究

Agrobacterium-mediated phyB gene transfer in Poncirus trifoliate

以枳壳(Poncirus trifoliate(L.)Raf.)实生苗上胚轴为试材,利用农杆菌介导法将光敏色素基因phyB导入其中,建立了枳壳实生苗的遗传转化体系,共获得27株抗性植株,PCR检测结果表明其中有5株扩增出phyB基因的特异目的带;RT-PCR分析结果表明phyB基因在phyB3,phyB4株系枳壳基因组中能表达;Li-6400光合测定仪对转基因枳壳进行的光合特性测定结果显示,转基因枳壳光合效率高于对照;转phyB基因枳壳植株在形态上与对照也存在差异,表现出植株矮化、节间缩短、叶面积变小.

The phyB gene was transferred into the genome of Poncirus trifoliate in vitro with explants of epicotyl mediated by Agrobacterium. Total 27 strains with plant resistance plants were regenerated. The presence of specific targeted strips of phyB gene in the 5 regenerated plants was confirmed by PCR analysis. RT-PCR analysis indicated the regular expression of transgene phyB in two transgenic clones, phyB3 and phyB4 in omparison with the control plants, the phyB transgenic plants had higher net photosynthesis rate. All the regenerated transgenic plants, as compared with the untransformed clones, showed lower height, shorter interval between intemodes and smaller leaf area.