Abstract
摘要:為保持蝴蝶蘭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。本計畫前期以蝴蝶蘭原球體為材料,轉殖ACC合成酶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。經GUS活性組織化學染色分析及南方氏雜交分析呈正反應的蝴蝶蘭癒傷組織反義ACC合成酶基因轉殖植株,進一步培養長大以進行開花性狀分析。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酶基因,運用農桿菌媒介法轉殖入大白花蝴蝶蘭原球體及癒合組織,經檢定含外源基因之轉殖植株,經培育長大為小苗並開花,本年度將進行轉殖植株性狀分析,將觀察順反義基因對蝴蝶蘭花色之影響,進一步進行性狀評估分析,選取特異株作為育種雜交材料。今年度尚將進行異黃酮類黃色素生成基因aureusidin synthase基因之
Abstract: To prolong the shelf-life of Phalaenopsis flowers, antisense ACC synthase cDNA from
Phalaenopsis was transformed into Phalaenopsis protocorm-like bodies (PLB).
Transformants were continuously propagated in vitro by means of flower-stalk-node
culture. Putative transgenic plants regenerated from calli with antisense ACC synthase
gene were identified by Southern analysis. Transgenic Phalaenopsis will grow in the
greenhouse for observation of their phenotype. On the other hand, to increase the
variation of flower colors of Phalaenopsis, Phalaenopsis chalcone synthase cDNA was
constructed as sense and antisense gene for genetic transformation. After genes were
transferred into Phalaenopsis PLBs and calli, 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. cDNA of flavonoid biosynthesis related gene aureusidin synthase will be used
as the probe for screening genomic library. Sequence of promoter will be analyzed for
conserved cis-acting elements. Furthermore, the promoter will be constructed into
expression vector for activity analysis. 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 confirmed by polymerase chain reaction, Southern analysis and inoculation test.
Keyword(s)
蝴蝶蘭
細菌性軟腐病
類黃素
基因轉殖植物
Phalaenopsis
bacterial soft rot
flavonoid
transgenic plants