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標題: 冰花訊息傳導相關mcSNF1在鹽逆境下基因表現及蛋白累積之分析
Analyses of gene expression and protein accumulation of a signal transduction-related mcSNF1 in halophyte Mesembryanthemum crystallinum under salt stress
作者: 楊邡郁
Yang, Fang-Yu
關鍵字: mcSNF1;冰花
出版社: 生命科學系所
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mcSNF1 (sucrose non-fermenting 1) 是利用yeast two-hybrid的方式在耐鹽模式植物冰花 (Mesembryanthemum crystallinum L.)以鹽處理的cDNA library中篩選出與mcSKD1 (suppressor of K+ transport defect 1)有交互作用的基因之一,已知鹽誘導基因mcSKD1參與逆境下intracellular vesicular trafficking之過程,本論文之目的在分析mcSNF1在鹽逆境下之基因表現與蛋白累積的情形,以期對mcSNF1的作用機制有進一步的了解。SNF1蛋白最早在酵母菌中發現,參與糖類代謝並與酵母菌許多抗逆境相關反應,其在植物中的同源蛋白SnRK (SNF1-related protein kinase)蛋白則參與許多植物抗逆境反應例如缺水逆境以及與ABA相關之訊息傳遞過程。

The mcSNF1 (sucrose non-fermenting 1) was identified through the yeast two-hybrid screening of a halophyte ice plant (Mesembryanthemum crystallinum L.) library using mcSKD1 (suppressor of K+ transport defect 1) as a bait. The salt-inducible mcSKD1 protein has been studied in depth that is involved in the intracellular vesicular trafficking in this halophyte. The main focus of this thesis is to examine the gene expression and protein accumulation of mcSNF1, a mcSKD1 interacting protein, in order to access its function in the complex network of salt tolerance in ice plant. The SNF1 was first discovered in yeast that has been shown to play roles in the sugar metabolism and stress responses. The plant homolog SnRK (SNF1-related protein kinase) has also been suggested in participating the stress responses and ABA-mediated signal transduction pathway.
The expression of mcSNF1 at cellular level was examined by RT-PCR and the accumulation of mcSNF1 was examined by Western blotting using an anti-AKIN10 antibody that can specifically recognize mcSNF1. The results showed long-term salt stress induced the expression of mcSNF1 and the accumulation of protein also increased concurrently. As for short-term salt stress, it was found the distribution of mcSNF1 was changed by salt treatment. The re-localization from the cytosolic vesicles to the plasma membrane occurred within hours of salt stress suggesting mcSNF1 is involving in the stress-related signal transduction pathway.
The constitutive expression of mcSNF1 was observed in intact plants and the addition of salt did not significantly change the level of expression as well as protein accumulation. Immunostaining of paraffin sections of leaves and roots did not reveal any tissue-specific localization of mcSNF1. The results suggested that the regulation of mcSNF1 in intact plant mainly occurs at the post-translational level but not at the transcriptional or translational levels.
In conclusion, at the early stage of salt stress, the distribution of mcSNF1 moves from the cytosol to the plasma membrane suggesting this protein is involved in the stress signaling. The amount of mcSNF1 expression increases as the stress persists in order to adapt the prolonged stress environment.
其他識別: U0005-2508200615010400
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