Co-transformation of Bt-toxin, Heat Shock Protein, Superoxide Dismutase, and Catalase Genes into Chinese Cabbage (Brassica campestris L. ssp. Pekinensis) and Cabbage (Brassica oleracea L. var. capitata L.) via Agrobacterium Mediated Transformation
| Autor: | Yuan-Chun Yeh, 葉元純 |
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| Rok vydání: | 2003 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 91 Taiwan is located at the sub-tropic area. In summer, the high temperature and humidity cause very serious of plant disease and insect pests. Attempts had been made to co-transfer the Bt-toxin gene (bt), superoxide dismutase gene (sod62) of Chinese cabbage, catalase gene (cat78) of Chinese cabbage, and heat shock protein gene (hsp70) of tomato into the cabbage (Brassica oleracea L. var. K-Y cross) and the Chinese cabbage (Brassica campestris L. ssp. Pekinensis cv. Tropical Pride). The objectives of this study are to establish the technology of gene co-transfer and targeting the foreign proteins into the chloroplast by chloroplast transit peptide, and to study the possibility for improvement of Brassica vegetables with insect, disease, and stresses resistance by gene transformation. The regenerated plants were further examined by PCR, Southern and Northern hybridization. Several transgenic plants contained four, three, two and one genes in one plant were demonstrated. The co-transformation rate of two, three, and four genes was 15%, 6% and 4%, respectively. The results of PCR, Southern and Northern hybridization indicated that the bt, hsc70, sod62 and cat78 genes had been inserted into the genomes of Chinese cabbage and cabbage, and expressed. The result of isozyme profiles assay revealed that the transformed SOD62 and CAT78 were present in the transformed plants. The SOD and CAT activities in leaves and chloroplast of transformed plants were higher than those of the controls. Increases in amounts of HSC70 protein were found in the transformed plants as analyzing by western blot hybridization. Preliminary results of bioassay indicated that the increases in tolerance to high temperature (42℃, 4 days), SO2 (800 ppb, 3 days), and insect resistance (Plutella xylostella) were found in the transgenic plants. This study demonstrated that two, three and four foreign genes could be co-transferred into one Chinese cabbage or cabbage plants simultaneously via Agrobacterium mediated transformation. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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