Transformation Of Ac/Ds Into Tomato And Overexpression Of Glucanase And Defensin Genes In Transgenic Tomato Enhances Resistance To Ralstonia Solanacearum

Tomato has been considered as a model for genomic study of Solanaceae because of its difference to Arabidopsis thaliana and rice in biology phenomena, its smaller genomic size (950 Mb) and powerful genetics groundwork. Construction of insertion mutant pool and function genome research will greatly devote to gene clone with important agriculture character and the related gene with independent knowledge property right. The gained mutants and their function genome research will supply theory and technique references for the study of the other vegetables and fruits, especially for berry fruit like Citrus and grape.The aim of this thesis was to generate a population of T-DNA insertion mutants via Agrobacterium-mediated genetic transformation using the constructed activation tagging into Micro-Tom and M82. With the identity and analysis of mutants, we expected the related genes would be isolate and clone in the coming work. The main results obtained were as followed:1. Seven activation tagging constructs were built promoted with CaMV 35S and Ac self-promoter, and with GFP, Lc, GUS as reporter genes.2. The constructs were transformed into Micro-Tom via Agrobacterium-mediated, and the system was optimized. The transformed tomato population was gained and the mean transformed efficiency was 56.7%. The transformed efficiency of pAcGFP was the highest construct, which reached 49.21%. In the process of GUSL transformation, no any transformed plant was gained because of the callus without differentiation. The assisted Ac/Ds insertional population with M82 was also built.3. GFP gene activation, diploid plants and molecular detection were conducted with the transformed plants. The results showed that GFP gene was interknitted into tomato genome. Expressions of GFP gene possess tissue and genetic material specific. The transformed plants were diploid plants, which was the same as the wildness Micro-Tom. Basta resistance experiment indicated that the average T-DNA inserted sites were 1.8 as the average inserted sites were 2.2 through southern blotting. Analyzed with southern blotting, about 26.0% among T1 population with one inserted site and 51.0% with two inserted sites, the average inserted T-DNA sites were 2.2. It showed the similar results as that by Basta analysis.4. The excision frequency of Ds element in cross-derived population was evaluated with...