| Summary: | 碩士 === 國立宜蘭大學 === 園藝學系碩士班 === 98 === Low pressure gene gun GDS-80 was designed and manufactered by local company and applied for it’s world patents. Currently the low pressure gene gun GDS-80 is used in transgenic-related research of animal system, and have established different bombardment conditions in cells, tissue of animal. However, in contrast the bombardment conditions for GDS-80 application in plant cells are undefine. One of the purpose of this study was to prove the general application of GDS-80 in plant tissues. By using the pCAMBIA1301 plasmid, which contains GUS gene as reporter gene, GDS-80 low pressure gene gun was applied to 6 different plant tissues with various pressure levels and distances. The results showed, by GUS histochemical observation, GDS-80 can successful transfer and express foreign gene in plant tissues, which confirmed the application of GDS-80 in gene transfer-related research for different plant cells. The study also obtains parameters for the best GUS expression in the 6 different plant tissues bombarded by GDS-80 low pressure gene gun. On the other hand, orchid is one of the economically important flowers in Taiwan. With the prosperous progress of international trade and biotechnology, the novel cultivars of orchid are actually under copycat behavior from unauthorized breeders and growers. We successfully developed a novel method for identification of novel cultivars in orchid. By introducing a short length foreign DNA sequence (27bp) as molecular marker via GDS-80 low pressure gene gun into PLB of orchid Blc. Tristar Beauty Beauty × Bc. Little Marmaid and P. Trne Lady × P. Borther Silk. Stable integrations of the 27bp foreign DNA sequence were confirmed by PCR and DNA sequencing of the putative transgenic orchids. The data also indicate that orchids Blc. Tristar Beauty Beauty × Bc. Little Marmaid and P. Trne Lady × P. Borther Silk have a certain positive transgenic regenerants without antibiotic selection performed in GDS-80 bombarded PLB of these two orchids. This experiment prove that plant variety rights identification can be established by introducing foreign gene marker as short as 27 bp.
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