Studies On Salt Tolerance And Drought Resistance Of Tomatos With Transgenic Technology

In this study, the drought tolerance gene Force is a transcription cofactors carrying the AT-hook DNA mutation, which was detached from Arabidopsis. The Force was under going a gene recombination and connection to the expression vector pBIN19, then agrobacterium-mediated tomato leaf blisk transformation method was used to obtain transgenic tomato plants. Through the whole genetic transformation process of drought tolerant gene Force in Money maker, we conclude main research results as following:1. Force was successfully reconnected to the expression vector pBIN19. The recombinant construction plasmid was storaged into the bacterial strain of Agrobacterium LAB4404 through shock transformation method, obtaining the engineering Agrobacterium with gene; extracting DNA from plasmid gained previously by PCR, and through XbaI, SmaI double-digested plasmid DNA, dual authentication results show that the gene Force has been inserted into the pBIN19 vector, moreover, it is indicated that the expression vector and engineering agrobacterium containing gene Force has been successfully constructed, which was necessary for the experiments.2. Learning from the experience of previous studies, an optimizatal efficient tomato transformation system was successfully built. The importance of this system has been greatly reflected from successful construction the plasmid vecto and acquirement of T0 generation of transgenic plants, all of which need the efficient tomato genetic transformation system, or else they can not be achieved. The rate of callus and budding was significantly increased by the use of growth hormone Trans-zeatin. Continuous screening on the transgenic seedlings was carried through antibiotics Tim, and the suitable selection pressure of hygromycin Hyg for Money maker was determined 5mg / L. Also, using hormone IBA in the rooting stage for GM greatly increased the number of positive transgenic seedlings; thereby reduced the workload of post-test identification and cultivation management.3. 16 T0 generation of transgenic seedlings with salt and drought tolerant gene Force were obtained, and the T1 generation seeds were harvested, which is the basis for getting excellent breeds with stable genetic characteristics of drought tolerance next. Within the genotypes of transgenic plants, it was confirmed that Force in tomato genome has been integrated by PCR identification, and RT-PCR analysis results show that the Force gene into tomato T0 generation conveys different levels of expression. Finally, Southern blot hybridization detected that, after cutting genomic DNA with a single SmaI, exogenous DNA in the tomato genome have different insertion sites, proving that the target gene Force has been inserted and integrated into the tomato genome.Within the phenotype of transgenic plants, most of plants pocess vigorous branches and leaves, tall and sturdy plant type, with many tussocky, wrinkle clusters and large leaf blade, and this is consistent leaf physiological characteristics of arabidopsis plants with Force gene.