| Summary: | 碩士 === 國立中興大學 === 園藝學系 === 91 === The results of increasing human population and scientific technology bring about the ruining of environment as well as enlarging the injured area. For example, greenhouse effect, acid rain and ozonosphere spoil etc. have resulted in a worldwide hurt for the nature, including Taiwan, where can’t be separated from the global weather change. In addition that Taiwan’s summer is extremely hot and humid, torrential rain, and drought stresses are speeding up the propagating the disease and destructive insects for crops. Therefore, breeding the vegetables of tolerates stress is an important direction. And using the transgenic technique is an effective way to improve the ability of tolerating the stress. The heat shock proteins cognate (HSC) is relation with tolerating high temperature, Bacillus thuringiensis (Bt) has the effect to kill insects, and the superoxide dismutase (SOD) and catalase (CAT) has the ability to decrease the free-radicle and tolerating the stress. In this study, hsc, bt, sod, and cat genes were co-transferred into cabbage (Brassica oleracea L. var. capitata L.) via Agrobacterium-mediated transformation.
The purposes of this study are to establish a co-transformation system of genes via Agrobacterium-mediated transformation and an explants regeneration model, to study the possibility of increase in the resistance to insects and environmental stress by gene transformation, and to explore the role of chloroplast transit peptide (TP) in delivering transformed proteins in the chloroplast.
The results of PCR, Southern blot, and Northern blot hybridization indicated that hsc70-2, bt, sod, and cat genes had been transformed into the chromosome of transformed cabbage and expressed the mRNA. The results of western analysis confirmed that the significant amounts of HSC70 were presented in the co-transformed plants than those of non-transformed plants. Moreover, the results of Western blot and SOD and CAT activity analysis showed that the expression of four transformed genes was higher in the co-transformed plants harbored with the TP than those of without TP. The results of bioassay showed increases in the resistance of heat, SO2, and insect (Plutella xylostella) in co-transformed cabbage. A better performance in the resistance to insect and environmental stresses were found in co-transformed plants harbored with the TP than those of without TP.
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