控制文心兰花瓣表皮细胞相关MYB基因表现之分析.doc

控制文心蘭花瓣表皮細胞相關MYB基因表現之分析 Analysis of MYB gene expression related to petal epidermalcell shape in Oncidium orchids 研究生：陳麗琦 Li-Chi Chen 指導教授：陳福旗 Fure-Chyi Chen 【摘要】 控制金魚草與矮牽牛花瓣表皮細胞相關基因所編碼之蛋白質屬於MYB基因家族，當基因突變時，會影響花朵表皮圓錐狀細胞的型態。本研究之目的為探討自文心蘭選殖的MYB33基因與花瓣表面突起或花色之關係。利用文心蘭（Oncidium Gower Ramsey）野生型與突變株Oncidium Gower Ramsey ‘Sunkiss’和白花突變株（alba mutant）進行分析。藉由型態觀察指出唇瓣表皮圓錐狀細胞開始突起的時期為小於0.5 cm的花苞。RT-PCR分析顯示MYB33基因主要表達於小於0.5 cm的花苞中，和唇瓣表皮圓錐狀細胞開始突起的時期相同，此時期恰為唇瓣類胡蘿蔔素開始累積的階段。在表皮細胞型態觀察，可發現相較於葉片平坦表皮細胞，花表皮為圓錐狀突起的細胞。在各組織部位的RT-PCR分析結果也顯示，除了白花突變株外，MYB33基因以花的表現量最高，由上述推測文心蘭MYB33基因可能參與表皮圓錐狀細胞的發育，亦可能參與表皮細胞中類胡蘿蔔素的累積。為了解文心蘭MYB33基因與花朵發育及花色表現時序間是否具有相關性，構築正義表現載體、RNAi干擾載體與OncMYBp::GUS啟動子表現載體，以農桿菌轉殖於菸草與文心蘭中。以GUS活性之組織化學染色分析確認構築之載體已成功轉入菸草、文心蘭與蝴蝶蘭基因組中，待其發育成不定根與不定芽，便可馴化出瓶，進一步進行轉殖後代分析。 文心蘭，MYB33基因，圓錐狀細胞，花瓣，轉基因植物The genes controlling the morphology of the petal epidermal cells in Antirrhinum and Petunia encode proteins belonging to the MYB gene families. The purpose of this study was to investigate the relation of MYB33 gene in Oncidium and the development of floral epidermal cells or the floral colour. The formation of conical cells in petals of the wild type and mutants occurs at 0.2-0.5 cm flower bud stages. Temporal expression of the MYB33 gene relative to different stages of flower development were analysed by RT-PCR. The MYB33 transcript level was highest in 0.2-0.5 cm flower bud stage, which also coincides with the occurrence of morphogenesis of epidermal cells and accumulation of carotenoids in labellum. The epidermal cells of peduncle are conical compared with epidermal pavement cells in leaf. Expression was also detected in peduncle except the alba mutant; the peduncle was higher than other tissues. We suggest the MYB33 gene may control the development of conical epidermal cells in petals in Oncidium. To understand the relation between floral development and floral colour, sense, RNAi and promoter in vector have been constructed, and transformation into tobacco and Oncidium by Agrobacterium infe