2011-04-202024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/684173摘要：為減少抗生素篩選基因使用之疑慮，本研究利用非抗生素抗性基因水稻 EPSP 合成&#37238;突變基因為篩選標誌，轉殖至蝴蝶蘭癒傷組織，配合Transposon系統移除篩選標誌基因。今年度之工作項目包括：1. 轉殖菸草比較蘭花剔除篩選標誌轉殖載體發生轉位情形。2. 蝴蝶蘭細胞經轉殖後，持續繼代培養，以取得均質的蝴蝶蘭轉殖細胞系。純化之轉殖蝴蝶蘭細胞系，將以聚合&#37238;連鎖反應及南方氏雜交分析，進行轉位作用之分子驗證。3. 雙抗病毒RNA干擾構築，將結合剔除篩選標誌通用載體，以農桿菌轉殖法進行蝴蝶蘭癒傷組織之基因轉殖，經非抗生素篩選取得純化之轉殖細胞系並再生後取得之轉殖株，將進行病毒之接種試驗。<br> Abstract: In order to achieve marker-free transformants, transposon system was chosen as maker-free strategy and the mutated rice EPSP synthase gene was used as selection marker which was resistant to herbicide glyphosate. The recombinant construct will be transformed into Phalaenopsis calli via Agrobacterium-mediated transformation. Transformed calli will be analyzed by polymerasechain reaction and Southern analysis to verify transpositionin Phalaenopsis and tobacco. Plasmid encoding double virus resistance gene against CymMV and ORSV will be constructed into markerfree system and transformed into Phalaenopsis calli. To enhance resistance to a soft-rot disease, pelE gene from Erwinia chrysanthemi driven by salicylic acid-inducible PR1a promoter will be transformed into Phalaenopsis calli and inoculation test will be accomplished.蘭科剔除篩選標誌系統果膠分解&#37238雙重抗病Orchidaceaemarker-free systempectate lyasedouble resistance