2004-01-012024-05-15https://scholars.lib.ntu.edu.tw/handle/123456789/662515摘要：花型優良、高價位的蝴蝶蘭，可作盆花及切花，但對乙烯相當敏感，且具有自動催化乙烯之特性，為保持良好之貯運品質以及瓶插壽命，本計畫選殖乙烯生合成相關基因，運用蝴蝶蘭轉殖系統，希望減低或延遲乙烯的大量產生，控制蝴蝶蘭切花之老化過程，以保持切花的新鮮度，將對蘭花產業將有甚大的經濟效益。本計畫前期轉殖ACC合成&#37238;反義基因所得到的品質優良轉殖蝴蝶蘭，本年度將繼續進行人工授粉、無菌播種及花梗芽插等組織培養繁殖。另一方面，為創新蝴蝶蘭之花色，增加蝴蝶蘭之花色變異，以有益於開拓市場，滿足國內外市場的需求不同與消費者普遍喜愛新奇花色的心態，故本計畫配合轉殖技術，將順、反義苯基苯乙烯酮合成&#37238;基因，運用農桿菌媒介法轉殖入大白花蝴蝶蘭原球體，經檢定含外源基因之轉殖植株，經培育長大為小苗，本年度將繼續以南方氏雜交分析驗證轉殖成功率。轉殖植株經培養長大開花，將觀察順反義基因對蝴蝶蘭花色之影響，詳細調查轉殖植株性狀，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，並利用已建立之蝴蝶蘭癒傷組織懸浮培養系統，以基因槍法及農桿菌共培養法，進行癒傷組織順義及反義苯基苯乙烯酮合<br> Abstract: ACC synthase cDNA from Phalaenopsis was constructed in opposite orientation as antisense plasmid for genetic transformation to prolong the shelf-life of cut flowers of Phalaenopsis. 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. 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 via particle bombardment or Agrobacterium-mediated method 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 antise ACC synthase, sense and antisense chalcone genes. The putative transformants are continuously selected by antibiotics in the media and regenerated into plantlets this year. Furthermore, we will construct pectate lyase gene pelE-1 from Erwinia chrysanthemi into the vector for Agrobacterium-mediated transformation and use the recombinant plasmid to generate transgenic Phalaenopsis which are resistant to bacterial soft rot disease.基因轉殖類黃素基因默化細菌性軟腐病蝴蝶蘭gene transformationflavonoidgene silencingbacterial soft rotPhalaenopsis