农杆菌介导菊花双菌共转化法中部分影响因素的研究

用XhoI酶切质粒pCAMBIA1301自身环化后得到T-DNA区只含GUS基因的重组质粒pCAMBIA1301-GUS,通过液氮冻融法将质粒pCAMBIA1301-GUS,pCAMBIA1300转入农杆菌LBA4404和EHA105中,并对菊花进行叶盘共转化实验,根据共培养3d后叶片中GUS的瞬间表达及其稳定共转化率,测定了不同农杆菌菌株搭配及其不同浓度配比对共转化效率的影响,结果表明：在两种农杆菌菌液浓度比为1：1时,其中EHA105/pCAMBIA1300与EHA105/pCAMBIA1301-GUS组合,共转化效率要高于其它菌株的组合,而在EHA105/pCAMBIA1300与EHA105/pCAMBIA1301-GUS组合中,两种菌液浓度比为1：2时共转化效率最高.

A Study on Several Factors Influencing Co-transformation of Chrysanthemum with Two Agrobacterium Strains

The plasmid pCAMBIA1301 was digested by XhoI and self-linked to get the recombinant plasmid pCAM- BIA1301-GUS,pCAMBIA1300 was transformed into Agrohacterium LBA4404 and EHA105 using a freeze-thaw method, the effects of either the combinations or the densities of the Agrohacterium strains on co-transformation efficiency were tested according to both the GUS transient expression in leaves and the stable co-transformation efficiency.The results showed that the combination with Agrohacterium EHA105/pCAMBIA1300 and EHA105/pCAMBIA1301-GUS achieved higher co-transformation efficiency than that of the other bacterium combinations when the same density of these two haeteriums was used to infect the leaves.Additionally,when the concentration proportion of the two Agrobacterium EHA105/pCAMBIA1300 and EHA105/pCAMBIA1301-GUS was adjusted to 1.2,the co-transformation efficiency was higher than that of the other concentration proportion.