The Transformation Of Malus Robusta With IRT1

In this study, Agrobacterium-mediated transformation system that was on the basis of the regeneration system, developed in our lab, was used to introduce the IRT1 gene into the Malus robusta, and several new transgenetic apple clones were obtained. The application of SAAT (Sonication-assisted Agnobacterium-mediated transformation) on tobacco and Malus robusta was discussed in this research. The results were as follow: .1. The method of SAAT could obviously increase the transformation efficiency on tobacco. Comparing with the rates of transient-expression which were gained by Agrobacterium-mediated transformation system, it is found that assisting with sonication in the progress of Agrobacterium-mediated transformation can get 100% transient-expression of gus gene, and the rate of area of blue spots was more than 50%. In the progress of the transformation of tobacco by SAAT, both the time and the power of sonication were the main factors that affected the efficiency of transforming. The optimal treatment was what was treated by 40 second in the power of 80%. In addition, the period of treatment also affected the transformation's efficiency. Assisted with the sonication after submerging by Agrobacterium for 4 min and 40 second was better than the other two treatments.2. SAAT was introduced into the transformation system of Malus robusta in the basis of which applied by tobacco in this study. The results showed that feasible treatment of sonication can also raise the efficiency of transforming of apple, and the trend of the affection of the sonication was the same as the one of tobacco. The results showed that the optimal treatment was the one that assisted with the sonication 20 second in the power of 70W after submerging by Agrobacterium for 4 min and 40 second, and through the treatment, the transient-expression of gus gene was 60% and the rate of injury of explants was lower than 23.3%. Besides, different kinds of explants which were treated by the sonication were also researched in this study. The results showed that the best kind of explants were the leafstalks, and compared with the other kinds of explants ,such as leaves and stalks, it can gain higher rate of transient-expression of gus gene with lower rate of injury.3. In this study, two kinds of strains were used to transform the Malus robusta:...