The Construction Of Plant Expression Vector And Expression Analysis Of TFT1、TFT10Genes In Tomato

The14-3-3protein, highly conserved in all eukaryotic cells, is an important regulatory protein. It plays an important role in the growth, signal transduction and other life activities of cells. In tomato,12isoforms have been found. The present evidence showed that14-3-3protein can bind to the SPS (sucrose-phosphate phosphatase) which is an improtent enzyme of sucrose metabolism in many crops, in turn altering its activity. In order to explore the relationships between the TFT1/TFT10genes and sucrose metabolism, Q-PCR was performed to analyse the expression patterns of TFT1and TFT10in cultivar tomato Micro-Tom fruits and wild tomato chmielewskii fruits. A regeneration system of wild tomato was established, and plant expression vectors of TFT1and TFT10were constructed and transformed into Agrobacterium EHA105. The main results were summarized as follows:1. Q-PCR expression analysis showed that TFT1and TFT10genes were expressed both in cultivated tomato and wild tomato fruit, but the expressions of them were different. Hexose-accumulating tomato Micro-Tom showed low TFT1expression at all stages with a small increase at the end of fruit development. Sucrose-accumulating tomato Chmielewskii showed low TFT1expression in immature fruit with a high increase at the end of fruit development, much greater than hexose-accumulating fruits.Throughout the development of fruits, hexose-accumulating fruits expressed the more TFT10than sucrose-accumulating fruits. The hexose-accumulating tomato Micro-Tom showed that the expression of TFT10gradually increaseed during fruits development. On the other hand, TFT10experssion remained low in the sucrose-accumulating tomato Chmielewskii at all stages of fruit development. This indicated that TFT1and TFT10might be involved in the regulation of sucrose metabolism in tomato.2. Established a regeneration system of wide tomato.The Optimum medium to induce the generation and differentiation of callus was screened out by using of cotyledon and hypocotyls as explants and the MS medium as the basic mediumin in this experiment, which were added in different kinds of plant growth regulators. The results showed that the buds frequency of cotyledon explants was much higher than hypocotyls; To cotyledon explants the best medium for callus induction was MS+0.5mg/L IAA+2.0mg/L6-BA, and the ratio of induction was100%; To hypocotyls explants the best medium for callus induction was MS+0.2mg/L IAA+2.0mg/L6-BA, and the ratio of induction was98%; The best medium for rooting was MS+1.Omg/L IAA, which the ratio of rooting could be100%.3. The plant expression vectors of TFT1and TFT10were constructed and introduced into Agrobacterium (EHA105). These works provided experimental foundation for further functional researches of TFT1and TFT10genes.