| Summary: | 碩士 === 國立臺灣大學 === 生命科學系 === 104 === Sinningia speciosa is a popular houseplant because of its big flower with a remarkable diversity in colors, patterns and shapes. S. speciosa has a small genome size (300 Mb), short life cycle, self-compatible, easily propagated in tissue culture therefore is emerging as a model plant for flower development studies. However, a reliable genetic transformation system is not available in S. speciosa. To this end, the Agrobacterium mediated transformation and particle bombardment transformation were tested in this study. Transient GUS expression assay showed that 3 days pre-culture of three weeks old seedlings on medium supplied with 1 ppm BA and co-culture for 5 days with Agrobacterium strain EHA105 achieved an overall transient transformation rate of 78.3%. Under these optimized conditions, the regeneration rate is 17.2 % and the transformation rate is up to 2.1 %. Another approach is particle bombardment transformation for optimizing genetic transformation system. In GUS transient assay, it was found that under helium pressure 900 psi, at distance 6 and 9 cm displayed the transient transformation rate of 58.1 % and 21.6 % respectively. The transformation efficiency of two approaches demonstrated that Agrobacterium-mediated
transformation is better than particle bombardment transformation. Because callus grows rapidly and regenerate easily, it serves as a good material for transformation. I also successfully induced embryogenic callus with 0.1 ppm 2, 4-D and 2 ppm BA plus 25 or 50 mM sorbitol in the medium. Callus transformation rate will be tested further. This study optimized the transformation protocol for studying gene regulation and gene function in S. speciosa.
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