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標題: 文心蘭中調控開花時間相關基因之選殖與特性分析
Molecular Cloning and Characterization of Genes Regulating Floral Initiation and Formation from Orchid (Oncidium Gower Ramsey)
作者: 邱怡芬
Chiu, Yi-Feng
關鍵字: Oncidium;文心蘭;Floral Initiation;LFY;開花基因
出版社: 生物科技學研究所
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高等植物開花過程是由營養組織轉變到花分生組織,進而進入一連串花之形成和花器發育過程,最終構成一朵完整的花。本研究之目的是針對調節文心蘭開花時期相關基因之選殖、功能性分析及應用,並進一步去了解如何達到提升文心蘭花卉品質及產量的目標。由文心蘭中選殖到三個與開花相關基因包括LEAFY (LFY)、MADS Box gene OMADS10 和 OMADS11,三者均是花分生組織形成及發育重要的調控者。OnLFY,序列上跟阿拉伯芥中的LFY具有同源性,由467 個胺基酸所組成並且和水稻的OsRFL有58%的相似性。RT-PCR分析顯示出OnLFY 高度表現於初期的花分生組織之原基(primordia)。大量表現OnLFY可促進阿拉伯芥野生種提早開花,OMADS10 相似於阿拉伯芥中A群MADS Box基因的 AP1, RT-PCR分析結果顯示此基因於所有的發育時期與器官皆可偵測到其表現,而OMADS11 相似於阿拉伯芥中E 群MADS Box 基因的SEP1/2,分析結果顯示OMADS11只表現於文心蘭中花器的部分。在35S::OMADS10 轉基因阿拉伯芥中觀察到整個植株會有提早開花並且有終結花(terminal flowers)產生的現象,而35S::OMADS11轉基因植株中會有植株矮小,產生腋生葉,極度早開花並且失去無限花序(inflorescence indeterminacy)等現象。本研究結果証明了在文心蘭中OnLFY 、OMADS10 和 OMADS11這三個基因在調控開花啟始及發育形成,皆扮演著重要的角色。未來將把基因大量表現於文心蘭,並利用RNA 干擾(RNAi)之方式阻斷基因表現,以便作更進一步功能性探討。

Flowering in higher plants involves the transition of vegetative development into floral initiation and formation and results in the production of flowers. The goal for this study is to clone and characterize orchid (Oncidium Gower Ramsey) genes regulating flowering time for further practical application of these genes in floral quality improvement. LEAFY (LFY) and MADS Box gene APETALA1 (AP1) are specific meristem identity regulator in controlling floral transition in plant. We report here the isolation of OnLFY, the orthologue of LFY from O. Gower Ramsey that encods a 467 amino acids protein and share 58% identity to rice OsRFL. RT-PCR analysis indicated that OnLFY was expressed in young flower primordia. Ectopic expression of OnLFY promoted flowering time in transgenic Arabidopsis. To investigate the flower formation of orchid, two MADS box genes, OMADS10 and OMADS11, were isolated from O. Gower Ramsey and functionally characterized in this research. OMADS10 showed a high similarity to the A function MADS Box gene AP1. RT-PCR analysis indicated that OMADS10 was expressed in all the organs and developmental stages tested. OMADS11, with high similarity to the E function MADS Box genes such as SEP1/2, was only expressed in flower and in all different floral parts. 35S::OMADS10 transgenic Arabidopsis plants flowered extremely early and produced terminal flowers. Transgenic Arabidopsis plants ectopically expressing OMADS11 also showed novel phenotypes resembling 35S::OMADS10 plants by significantly reducing plant size, producing curly leaves, flowering early, and losing inflorescence indeterminacy. Our result provided evidence to show that OnLFY, OMADS10 and OMADS11 play multiple roles in regulating both floral transition and formation in orchid. The RNA interference technology was applied to generate transgenic mutants to further investigate the function for these genes.
其他識別: U0005-0608200714572400
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