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標題: | 靜默矮牽牛中SEPALLATA基因造成花朵器官的轉變 Effects of floral organ conversions caused by silencing of petunia SEPALLATA |
作者: | Yu-Hang Lu 陸禹涵 |
指導教授: | 盧虎生 |
共同指導教授: | 陳仁治 |
關鍵字: | 矮牽牛,VIGS,SEPALLATA,FBP9,FBP23,萼片葉化, petunia,VIGS,SEPALLATA,FBP9,FBP23,leafoid sepal, |
出版年 : | 2011 |
學位: | 碩士 |
摘要: | 近年來ABCE模型已廣泛的解釋不同物種中花朵器官的形成,而花朵器官的發育乃由A-class (AP1及AP2)、B-class (AP3及PI)、C-class (AG)和E-class (SEP)四群基因的交互作用所調控。而E-class基因在阿拉伯芥當中擁有功能上的保守性。單一SEP基因的突變並不會造成外表型的改變,而阿拉伯芥的四個SEP中,在有三個或四個基因發生突變時,則分別使所有花朵器官轉變為花萼和葉片。在矮牽牛當中目前已發現E-class基因有FBP2、FBP4、FBP5、FBP9、FBP23與MADS12六個,但是僅僅是FBP2與FBP5的突變即可在花瓣、雄蕊及心皮中觀察到花朵器官的改變。並且從序列的分類來看,矮牽牛中具有一只在茄科植物中存在的次類群(FBP9和FBP23)。因此懷疑在矮牽牛中的E-class基因已有不同的特化與分工。為了瞭解矮牽牛中E-class基因的功能保守性是否與阿拉伯芥的E-class基因有所不同,本研究觀察擁有葉化萼片的實驗品種M1與擁有正常花萼之實驗品種W115做比較,分析其花朵四輪構造中E-class基因表現量的差異;以及使用利用Tobacco Rattle Virus (TRV) 誘發之Virus-Induced Gene Silencing (VIGS) 的技術,利用FBP23的保守性片段、部分保守性片段與專一性片段,靜默矮牽牛當中E-class的基因,觀察花朵外表型的改變與E-class基因RNA表現量間的關係。結果顯示M1萼片中除了MADS12外的E-class基因表現量很低。在接種三種含FBP23片段之TRV誘發VIGS之花朵的結果顯示,單獨靜默FBP23並沒有明顯的外表型,而在接種含FBP9和FBP23專一片段的TRV植株中,觀察到FBP9表現量提升,是否與萼片葉化有關目前還無定論。變異的花朵中,瓣化雄蕊的FBP2、FBP9與MADS12其表現量與正常花瓣類似,相較於M1中花瓣與雄蕊的差異,可能為FBP4、FBP5與FBP23共同的調控雄蕊與花瓣的決定。 ABCE model has been widely interpreted in flower organ formation of different species. In recent years, According to the model, flower organ formation is determined by interaction of four groups of genes, A-class (AP1 and AP2), B-class (AP3 and PI), C-class (AG) and E- class (SEP) genes. In Arabidopsis, there is a conservative function in E-class proteins. No phenotypic change was observed in single SEP gene mutants while when tree or four Arabidopsis SEP genes were mutated, all flowers organs were converted into sepals and leaves respectively. In Petunia, six E-class proteins, FBP2, FBP4, FBP5, FBP9, FBP23 and MADS12 have been found, and it was reported that down-regulation of two genes, FBP2 and FBP5, petals, stamens, and carpels were converted into sepaloid organs. The phylogenetic tree indicated that one subgroup of E-class genes (FBP9 and FBP23) only existed in Solanaceae. It was suspected that there are specialization and different functions in the E-class genes of petunia. To find out the functional differences among Petunia E-class proteins, we compared transcript abundance of E-class genes of experiment strain M1, which has leafloid sepals, and W115, which has normal sepals. Moreover, we used Tobacco Rattle Virus (TRV) that contains a FBP23 cDNA fragment to inoculate petunia plants to induce gene silencing, and then analyzed the correlation between morphological changes of flower organs and transcript abundance of E-class genes. The results showed that most of the E-class genes were repressed expect MADS12 in sepals of M1 flowers. Flowers from plants inoculated with TRV containing specific FBP23 sequence fragment showed no obvious morphological changes. However, we observed that an increase of FBP9 expression and an reduction of FBP23 expression in a flower showed leafloid sepals, which was from a plant inoculated with TRV containing FBP9 and FBP23 specific sequence fragments. In transformed flower, the expression pattern of FBP2, FBP9 and MADS12 in petaloid stamen is similar to normal petals. Compare with the petals and stamen of M1, that FBP4, FBP5 and FBP23 may be the co-regulators of petals and stamen identity. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/31407 |
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