Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/63277
標題: 文心蘭中與花型發育及開花時間調控相關基因之選殖、分析與應用
Cloning, Characterization and Agricultural Application of Genes Controlling Flower Organ Formation and Flower Transition in Oncidium Gower Ramsey
作者: 楊長賢
關鍵字: 應用研究;生物技術, 園藝
摘要: 
在台灣加入世界貿易組織後,發展觀賞花卉產業無疑在未來台灣的農業將佔上一席之地。而文心蘭(Oncidium Gower Ramsey)無疑就是重點花卉之一,因而國家對其之研究至為重視。一朵花由花萼、花瓣、雄蕊與雌蕊四部份構成。目前對花內器官的形成在分子基因層次上的研究已知是由一群基因之間共同調控的相互關係,而這些與花型相關的基因幾乎都是屬於含一個高保守區域MADS box domain的基因。本實驗室對文心蘭花型特徵改變及開花時間調控的研究深感興趣,已成功的由文心蘭中將屬於A、B、C、D與E基因群的MADS box基因全部選殖出,共有11個基因,分別名為OMADS1-OMADS11。在將這些基因轉殖入阿拉伯芥(Arabidopsis thaliana)及菸草等中大量表現後,的確造成轉基因植物花器形成或開花時間上程度不一的改變。這些結果對將來我們操控及改造文心蘭之開花時間及花型提供很大的信心。本研究將以進一步改進文心蘭之開花時間及花型為目的。首先要將這11個基因進一步轉殖入其他已具基因轉殖系統之觀賞花卉洋桔梗中作測試,由轉基因植株之表現形態更加確立這些基因之功能。接著將已構築好含這些基因cDNA正譯(sense)、反譯(antisense)、RNAi或各類缺失(truncated)之構築體(constructs)直接轉殖文心蘭,並分析轉殖植物性狀,以產生具有開花時間及重要花型改變之轉殖文心蘭。本計畫同時要進行的是由文心蘭中選殖更多的與開花時間及花器形成相關之MADS box基因,完成其RNA和蛋白質表現的測試,接著這些基因將被轉殖到阿拉伯芥、菸草及洋桔梗中作測試分析,最終將送人文心蘭中進行改造。另外因文心蘭栽培之最大瓶頸在於開花期太過集中,故透過生物科技之基因調控研發花期調節之技術實為重要,本實驗室已成功的由文心蘭中成功選殖出三個與開花時間相關的基因,OnGI, OnFT, 及OnFLD。因此本計畫預計完成將這些基因轉殖到阿拉伯芥、菸草及洋桔梗中進行分析,最終將送人文心蘭中進行開花時間的改造。本計畫同時亦要進行由文心蘭中選殖與應用其他與開花時間相關的基因,如CO, LEAFY等。本計畫中所有轉殖基因的文心蘭植株中若出現重要及顯著具市場價值的花型及開花時間的改變,則可立刻進行田間測試,並評估及推廣到市場。另外所有含本計畫選殖出基因之各式構築體可立刻用來做其他重要植物或作物轉殖及改良用。此外本計畫同時要繼續進行文心蘭開花前及開花期之subtraction cDNA library及相關Expression Tag Sequences (ESTs) library資料庫的建立。透過序列分析這些EST及特異表現之cDNA clones,不只可建立文心蘭的表現基因庫,提供將來整體文心蘭基因體計畫相關實驗如microarray等之運用,更可能從中直接選殖到與開花相關基因之同源基因(homologues)。這些基因可以和前述的基因一樣被轉入阿拉伯芥、菸草、洋桔梗中作測試,最終送人文心蘭中,以達到多方面改良文心蘭花卉的目的。

To face the effects potentially caused by attending WTO, the development and enhancement of flower industy is necessary for Taiwan』s agriculture. Oncidium Gower Ramsey, a species of orchid, has become one of the most important flowers in flower industy during past ten years. Flowers are composed of four whorls of organs, sepal, petal, stamen and carpel. Molecular and genetic studies have revealed that floral organ formation was controlled by a group of MADS box genes which contained a conserved N-terminal MADS box domain. To study the flower development in O. Gower Ramsey, eleven MADS box genes (OMADS1 to OMADS11) including all ABCDE functional genes, were isolated and characterized from O. Gower Ramsey in our laboratory. Ectopic expression of these genes in Arabidopsis thaliana or tobacco produced novel flowers contained the homeotic conversion of flower organs as well as significantly promoted flowering. These results revealed the possibility for using these genes in flower modification. In this project, constructs containing various forms of OMADS genes will be first introduced into plants such as Eustoma grandiflorum for further functional confirmation. Furthermore, these constructs will be transformed into O. Gower Ramsey and the phenotypes analyzed. Any novel phenotypes in flower shape and flowering time will be characterized, evaluated, and subjected into field try and flower market. In addition, to stusy the regulation of floweing time in O. Gower Ramsey, flowering time associated genes such as OnGI、OnFT and OnFLD have been cloned from orchid in our laboratory. In this project, these genes will be functional analyzed in Arabidopsis, tobacco and E grandiflorum through transgenic approach. Same as MADS box genes, constructs containing various forms of these genes will be transformed into O. Gower Ramsey. Interested flower phenotypes will also be subjected into flower industry. We also plan to clone more MADS box genes involved in flower transition and formation as well as other flowering time associated genes such as CO and LEAFY. Their RNA and protein expression will be analyzed. Constructs containing various forms of these genes will be functional analyzed in Arabidopsis, tobacco, E grandiflorum and O. Gower Ramsey through transgenic approach. Furthermore, subtraction cDNA and Expression Tag Sequences (ESTs) library will be constructed for O. Gower Ramsey. Genes specifically expressed before or after bolting will be sequenced and analyzed. Clones showed sequence homology to flowering time genes will be characterized and functional analyzed. All the genes cloned will be transformed into O. Gower Ramsey to generate interesting phenotypes. All useful phenotypes generated in this project will be used directly for agricultural application in controlling flower development in O. Gower Ramsey as well as in various plant species.
URI: http://hdl.handle.net/11455/63277
其他識別: NSC96-2317-B005-019
Appears in Collections:生物科技學研究所

Show full item record
 

Google ScholarTM

Check


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.