請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/35886
標題: 百合中三個調控開花起始之AP1groupMADSBox基因的分子選殖與功能分析
Molecular Cloning and Characerization of Three Lily (Lilium longiflorum) AP1 Group of MADS Box Genes in Regulating Floral Initiation
作者: 林怡君
Lin, Yi-Chun
關鍵字: Lilium longiflorum
鐵砲百合
AP1
MADS Box
Floral Initiation
開花起始
花器形成
出版社: 農業生物科技學研究所
摘要: 在植物中,任何花器的形成皆受到數種homeotic基因交互作用所調節。由於這些基因大都屬於會在N端轉譯出含有保守DNA鍵結區域的MADS box基因蛋白,顯示MADS box基因在花朵發育上扮演著重要的角色。除了參與花器形成外,在擬南芥中的AP1和AGL9之MADS box基因也參與了調節開花時間的啟始。為了研究百合開花之啟始過程,本研究從百合中選殖三個與AP1具有高度相似性的MADS box基因並做進一步的特性分析。LAP1-1可轉譯出一個含252個胺基酸的蛋白與水稻的AP1同源基因OsAP1-like有60%一致性和71%相似性。在MADS box區域方面,LAP1-1與OsAP1-like之間有90% (52/58)的一致性。而LAP1-2和LAP1-3 Δ cDNA皆缺乏5’端的MADS box區域,經由序列比較顯示LAP1-2比LAP1-3更像LAP1-1,此外,三者表現的特徵也很相似。這三個基因之mRNA在花苞、營養葉和花序分生組織皆可被偵測到。在花器中之表現與擬南芥中AP1的表現情形相似,在4個花器皆有表現。有趣的是,LAP1-1與LAP1-2在雌蕊及葉的部分可以觀察到有類似的高表現量。再進一步分析大量表現LAP1-1之轉基因擬南芥植株發現,有提早開花及產生捲曲葉的現象。在花器中有花萼轉型成雌蕊及花瓣轉型成雄蕊的變異。這些結果強烈的顯示LAP1-1的功能除了參與花的形成外,也參與了花的誘導。另外在蛋白質功能分析上,利用酵母雙雜合之交互作用結果顯示LAP1-2會形成homodimer並且會與LAP1-1形成heterodimer,而LAP1-1本身不會形成homodimer。將來透過更進一步分析LAP1-2與LAP1-3的功能,相信不但可以使我們更深入的了解MADS box基因調控花的轉型,而且也可以提供未來有效運用這些基因的策略。
The formation of any flower organs was regulated by the interaction of several classes of homeotic genes in plants. Since most of these genes are MADS box proteins which contained a conserved DNA-binding domain (the MADS box domain) at the N-terminus, indicating the central role for MADS box genes in flower development. In addition to the involvement in the flower formation, MADS box genes such as AP1 and AGL9 of Arabidopsis have also been thought to anticipate in the regulation of floral transition and initiation. To investigate the flower initiation in lily (Lilium longiflorum), three cDNA showed high homology to AP1 group of MADS box genes were isolated and characterized. LAP1-1, encodes a 252 amino acid protein that showed 60% identity and 71% similarity to rice AP1 homologue OsAP1-like. In MADS box domain, 90% (52/58) amino acids are identical or similar between LAP1-1 and OsAP1-like. cDNAs for LAP1-2 and LAP1-3 are partial sequences truncated with 5' MADS box domain. Sequence comparison indicated that LAP1-2 was more closely related to LAP1-1 than LAP1-3 was. In addition to the sequence similarity, the expression pattern for these three genes was also similar. Messenger RNA for these three genes was detected in floral buds as well as in vegetative leaf and inflorescence meristem. In flower, similar to AP1, they expressed in all four flower organs. Interestingly, similar high expression in carpel and leaves was observed for LAP1-1 and LAP1-2. Ectopic expression of LAP1-1 in transgenic Arabidopsis plants showed novel phenotypes by flowering early and producing curly leaves. Homeotic conversion of sepals to carpelloid and petal to stamen-like structures were also observed in 35S::LAP1-1. These results strongly indicated that the function of LAP1-1 is involved in flower formation as well as in floral induction. Yeast two-hybrid analysis indicated that LAP1-1 truncated with the MADS box domain is able to form heterodimer with LAP1-2. Interestingly, LAP1-2 is also able to strongly form homodimers. By contrast, LAP1-1 is unable to form homodimers. Further functional analysis for LAP1-2 and LAP1-3 should lead not only to a deeper understanding of the MADS box genes in regulating flower transition but also to provide useful strategy to manipulate these genes in the future.
URI: http://hdl.handle.net/11455/35886
顯示於類別:生物科技學研究所

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