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Study on the Transformation of Apple and Tomato with Apple Genes DHAR and MDHAR Mediated by Agrobacterium Tumefaciens

Author: WangPingPing
Tutor: MaFengWang
School: Northwest University of Science and Technology
Course: Pomology
Keywords: Ascorbic acid Transgenic DHAR MDHAR Tomato Apple
CLC: S661.1
Type: Master's thesis
Year: 2011
Downloads: 101
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Abstract


Ascorbic acid (Asorbic Acid, AsA) that vitamin C (Vc), the organism has important metabolic function and antioxidant effects. Improve the plant, especially the ascorbic acid content of fruit products, not only can increase the nutritional quality of plant products, the plant itself, but also enhance resistance to adversity. Plants, ascorbic acid in the role of ascorbate peroxidase and ascorbic acid oxidase, oxidized single DEHYDROASCORBATE. Single dehydroascorbate unstable again a single dehydroascorbate reductase (MDHAR), under into ascorbic acid, or no action of the enzyme metabolized into dehydroascorbate. The dehydroascorbate spontaneous hydrolysis into 2,3 - dione Angulo acid, or the use of reduced glutathione (GSH) to regenerate ascorbic acid as a reducing agent in the role of dehydroascorbate reductase (DHAR). Dehydroascorbate reductase, and single dehydroascorbate reductase plays an important role for the regeneration of ascorbic acid. Antioxidant in tomato leaves apples and tomatoes, the test material, the use of Agrobacterium-mediated transformation, and cloned into the DHAR the gene and MDHAR gene of Greensleeves apples and tomatoes translational research from Apple, get on the conversion system enzyme activities were measured. Provides a theoretical basis for the study of the ascorbic acid cycle of the DHAR and MDHAR two regeneration enzyme. The main findings are the following: 1 Greensleeves apple plantlets leaves were used to study different hormone combination of different leaf age, The different leaves placed its regeneration optimize Greensleeves Apple regeneration system. The optimum regeneration medium for the 6-BA5.0 mg / L NAA1.0 mg / L 30g / L sucrose, 7.0g / L agar (pH5.8). 2-3 blades, best leaf stage for 20 days subculture top fully extended the blade nearly axial end close to the medium than the far axial end close to the medium having a significantly higher regeneration rate. 2 apple leaf receptor using Agrobacterium-mediated transformation Vc the metabolic regeneration key enzyme gene transferred to Apple DHAR and MDHAR of Apple transformed plants. 3 tomato \The results showed that the regeneration rate is much higher than the cotyledon as explants following hypocotyl explant regeneration rate, the best regeneration medium MS 2.0 mg / L ZT 0.2 mg / L IAA 30 g / L sucrose 7.0 g / L agar (pH5.8), cotyledon regeneration rate of 99%. Hypocotyl regeneration rate can only reach 75%. 4 tomato cotyledons receptors, the apple the DHAR gene and MDHAR of genes transferred into tomato using Agrobacterium-mediated transformation, get the 12 tomato DHAR transgenic plants with two MDHAR transformed plants. GUS staining and PCR results showed that the target gene has been transferred to the DHAR and MDHAR of the tomato genome. The five pairs DHAR and MDAHR of transformed plants APX, CAT, DHAR, MDHAR, GR activity were determined. The activity of the transgenic tomato plants has increased the highest increased 2-fold.

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CLC: > Agricultural Sciences > Gardening > Fruit trees gardening > Pome > Apple
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