碩士 / 銘傳大學 / 生物科技學系碩士班 / 101 / Trehalose is a stable, non-reducing disaccharide,it is two glucose molecules constituted by 1,1 glycoside bond. Present in plants, animals, microorganisms widely, and has function to protect cells and protein structure. Trehalose plays an important role to helping plants defense abiotic stress, like heat, drought, chilling and salinity. Plants will produce trehalose to confrontation with abiotic stress. In plants, the trehalose synthesis requires two enzymes, trehalose phosphate synthase (TPS) and trehalose phosphate phosphatase (TPP), and its reactants are UDP-Glucose and Glucose-6-phosphate. In previous studies, there is a kind of trehalose synthase produce by Picrophilus torridus, it is Picrophilus torridus trehalose synthase (PTTS), this archea can resistant heat and low pH, and PTTS is the same, too, PTTS can use maltose as reactant to produce trehalose, so we want to build this system in Arabidopsis, and observing what effect will happens. We use Geteway Cloning system to construct PTTS gene into the pPZP200 expression vector, and then transform pPZP200/PTTS plasmid into Agrobacteria, we use agrobacteria mediated transformation to infection Arabidopsis and collect seeds (T0). Then sow seeds on 1/2 MS plate contain hygromycin to select transgenic plants (T1), collect seeds get homozygote (T2) and culture to mature do analysis and abiotic stress tests. Now we extracted genomic DNA, total RNA and Western blot, to make sure PTTS gene has expression. So far, all experiment can confirm that PTTS will be expression in transgenic plants, the transgenic plant is smaller than non-transgenic plant, and transgenic plants has stronger resistance in heat stress, it indicating that PTTS can improve the ability to resist heat stress in Arabidopsis.
| Identifer | oai:union.ndltd.org:TW/101MCU05111006 |
| Date | January 2013 |
| Creators | Yao-Jen Chang, 張耀仁 |
| Contributors | Chin-Ming Chiang, 江志明 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 48 |
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