Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/90089
標題: The NAC-like gene AtNACL14 Controls Plant Growth and Development by Regulating Gibberellin Metabolism pathway in Arabidopsis
阿拉伯芥NAC基因AtNACL14調控植物荷爾蒙吉貝素生合成路徑以參與及控制植物生長發育
作者: 陳宏翊
Hong-Ie Chen
關鍵字: 吉貝素;阿拉伯芥;gibberellin;NAC;arabidopsis thaliana
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Plant Cell, 19, 2776-2792.
摘要: 
NAC (for NAM, ATAF1, 2, and CUC2)基因為一群植物特有的基因群。大部分的NAC蛋白在其N端均含一個約為150胺基酸的高保守性DNA-binding domain以及NLS (localization signal sequence)序列。以阿拉伯芥為例,NAC蛋白參與許多植物生長和發育的過程,例如cell cycle的控制、生長荷爾蒙訊息的傳遞、花器的發育和頂端分生組織的形成等。目前在阿拉伯芥中,有一群NAC蛋白C端被預測出具有α螺旋的穿膜功能域,此功能域被認為與蛋白結合在膜上的能力有關。根據酵母菌的活性篩選分析的結果,推測AtNACL14在阿拉伯芥中扮演活化子的角色。異位表現AtNACL14全長的轉基因植物與野生行阿拉伯芥相比,無相異性狀,然而剔除穿膜區域的AtNACL14ΔTM轉基因植物,則呈現矮化與營養葉捲曲等性狀。藉由將AtNACL14融合GFP螢光蛋白的方式,來觀察其在次細胞層次的表現與分布位置,我們發現AtNACL14蛋白質會出現在內質網之膜上,反之剔除穿膜區域之AtNACL14ΔTM蛋白則會位在細胞核內,這個結果顯示AtNACL14蛋白是由內質網膜上切出然後進入細胞核中執行功能。經由生物晶片的分析,在異位表現剔除穿膜區域的AtNACL14ΔTM轉基因植物中,GA2ox1與GA2ox2被誘導表現,而GA20ox1及GA20ox相關的基因的表現則被抑制。進一步藉由即時定量聚合酵素鏈鎖反應分析,在吉貝素的生合成代謝路徑中,參與合成的酵素(GA20ox)被抑制,反之涉及代謝的酵素(GA2ox)表現量卻上升。此外,剔除穿膜區域的AtNACL14ΔTM轉基因植物的內生性吉貝素含量,相較野生型阿拉伯芥低。在低溫下AtNACL14會進入細胞核且GA2ox2表現量會上升,綜合上述,我們推測AtNACL14可藉由控制吉貝素的動態平衡來調控植物的生長發育。

AtNACL14 contained one highly conserved NAC domain in N-terminal region and a strong α-helical transmembrane motif (TM) in C-terminal region and has been predicted to be membrane-associated. AtNACL14 has transactivation capacity based on yeast transcription activity assays. This result indicated that AtNACL14 acts as a transcriptional activator. Transgenic plants ectopically expressing AtNACL14 was phenotypically indistinguishable from wild-type plants. Ectopic expression of AtNACL14ΔTM exhibited severe phenotypic alterations such as dwarfism and curled leaves in transgenic Arabidopsis. GFP+AtNACL14ΔTM (lacking a transmembrane domain) fusion proteins were accumulated in the nuclei of the Arabidopsis cells whereas the GFP+AtNACL14 fusion proteins were accumulated in the ER membrane and absent in the nuclei. These results revealed that AtNACL14 needs to be processing and releasing from the mitochondria to perform its function. Data of microarray showed that GA2ox1 and GA2ox2 were up-regulated but GA20ox1 and two GA20ox-related genes were down-regulated in 35S::AtNACL14ΔTM plants. Further analysis of the expression for genes in GA metabolism pathway indicated that most GA biosynthesis genes (GA20ox) were down-regulated whereas two GA deactivation genes (GA2ox) were up-regulated in 35S::AtNACL14ΔTM plants. In addition, levels of gibberellins in 35S::AtNACL14ΔTM plants were lower than that in wild type plants. Furthermore, 4℃ cold treatment caused the entering of AtNACL14 into nucleus and the up-regulation of GA2ox2. Our data suggest a role for AtNACL14 in controlling multiple plant developments by suppressing the GA biosynthesis in Arabidopsis.
URI: http://hdl.handle.net/11455/90089
Rights: 同意授權瀏覽/列印電子全文服務,2018-08-20起公開。
Appears in Collections:生物科技學研究所

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