2005-01-012024-05-15https://scholars.lib.ntu.edu.tw/handle/123456789/662516摘要：蝴蝶蘭花型優良價位高，但對乙烯相當敏感，本計畫應用生物技術，希望減低或延遲乙烯大量產生，以保持良好之貯運品質及瓶插壽命。本計畫前期以蝴蝶蘭原球體為材料，轉殖ACC synthase反義基因，得到的優良轉殖株，本年度將繼續繁殖；蝴蝶蘭癒傷組織轉殖株將進行再生、GUS活性組織化學染色分析及南方氏雜交分析。另一方面，為創新蝴蝶蘭之花色，以滿足國內外市場的需求，本計畫將順、反義chalcone synthase基因，轉殖入大白花蝴蝶蘭原球體，轉殖株經培育長大並開花，本年度將進行性狀分析，選取特異株作為育種雜交材料；為避免以原球體為轉殖材料容易發生鑲嵌問題，也利用蝴蝶蘭癒傷組織進行轉殖，轉殖後的細胞經以抗生素篩選後，經光照處理分化為類原球體並再生，以GUS活性組織化學染色分析及南方氏雜交分析確認後，將培養長大開花，觀察順、反義基因對蝴蝶蘭花色之影響，並進行性狀評估。今年度尚將進行黃色素生成基因之分離、定序及表現載體構築。另將進行轉殖軟腐病抗性基因pectate lyase基因pelE-1及pelZ之蝴蝶蘭癒傷組織的篩選與再生，期望培育出具有抗軟腐病能力的蝴蝶蘭品種。<br> Abstract: To prolong the shelf-life of Phalaenopsis flowers, ACC synthase cDNA from Phalaenopsis was constructed in opposite orientation as antisense plasmid for genetic transformation. After screened with antibiotics, the putative transformants were grown to flowering stage and the flowers were analyzed for the effect of inhibition on ethylene synthesis. According to the results of flower-life measurements, GUS staining and Southern analysis of genomic DNA from transformants has demonstrated the integration of foreign gene into the genome of transgenic plants. They are continuously propagated in vitro by means of flower-stalk-node culture. Putative transgenic calli with antisense ACC synthase gene will be continuously selected by antibiotics in the media and regenerated into plantlets this year. On the other hand, to increase the variation of flower colors of Phalaenopsis, Phalaenopsis cDNA encoding chalcone synthase was isolated and constructed into transformation vector as sense and antisense gene driven by CaMV 35S promoter and stopped by NOS terminator. After genes were transferred into Phalaenopsis genome using protocorm-like bodies (PLB), the putative transformants were selected with antibiotics and analyzed by Southern hybridization to confirm the success of transformation. The transformants with novel flower color will be used as breeding materials. Furthermore, to avoid creating chimeric plants generated from PLB after transformation, the application of calli as transformation target material and the system of suspension culture have been established and transformed with sense and antisense chalcone genes. The putative transformants will be continuously selected with antibiotics in the media and regenerated into plantlets this year. We will try to isolate genes involved in biosynthesis of yellow pigments and construct those genes into expression vectors. Meanwhile, bacterial soft rot disease resistant gene pectate lyase gene pelE-1 and pelZ from Erwinia chrysanthemi were transformed into Phalaenopsis via Agrobacterium-mediated transformation. The putative transformants will be continuously selected with antibiotics in the media and regenerated into plantlets this year.轉殖植物類黃酮細菌性軟腐病Transgenic plantflavonoidbacterial soft rot