頁籤選單縮合
題名 | 水稻OsPII基因的選殖及功能分析=Cloning and Functional Characterization of OsPII in Rice |
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作 者 | 林子光; 侯新龍; | 書刊名 | 嘉大農林學報 |
卷期 | 8:1 2011.02[民100.02] |
頁次 | 頁23-34 |
分類號 | 434.114 |
關鍵詞 | 水稻; PII蛋白質; 葉綠體; 基因過量表現; 氮素訊息; Rice; PII protein; Chloroplast; Gene overexpression; Nitrogen signaling; |
語文 | 中文(Chinese) |
中文摘要 | PII蛋白質在藍綠細菌、大腸菌及阿拉伯芥中被認為是擔任氮素訊息的感應子及細胞中C/N平衡的調節者。為了瞭解水稻細胞中的氮素訊息路徑,因此自水稻中選殖出與阿拉伯芥PII的同源基因,命名為OsPII。為了確定OsPII在水稻中是否也有類似上述的功能,我們構築了OsPII基因過量表現及OsPII-GFP的融合蛋白表現載體。經由農桿菌轉殖到水稻中,GFP的表現證明OsPII蛋白質位於葉綠體中,而過量表現OsPII基因的轉殖細胞系,在缺氮培養下,也會促進某些受缺氮誘導表現的半胱氨酸蛋白酶基因的表現。此外,OsPII過量表現的T1種子發芽後幼芽及根的生長速率較高於WT,推測可能過量表現OsPII將促進半胱氨酸蛋白酶基因的表現,因此更有效的分解胚乳中的儲藏性蛋白質,提供幼苗生長所需的氮素源。根據上述結果,我們推測水稻的OsPII蛋白質可進入葉綠體內,感測氮素訊息或氮素代謝的調控。 |
英文摘要 | In cyanobacteria, E. coli and higher plants, PII functions as a nitrogen sensor and plays a crucial role in regulating cellular carbon and nitrogen levels. To understand the nitrogen signaling pathway in rice cells, rice orthologue of Arabidopsis PII cDNA was cloned and designated as OsPII. To functional dissect the OsPII gene, UbiP:: OsPII and UbiP:: OsPII-GFP were constructed and transformed to rice via Agrobacterium-mediated transformation. Analysis of transgenic rice containing UbiP:: OsPII-GFP indicated that OsPII protein was a chloroplast-localized protein. Overexpression of OsPII gene in rice cells activates two cysteine proteinase genes, Oryzain and Oryzain, under nitrogen starvation condition. Moreover, the growth rate of seedlings in overexpressing lines was higher than WT after seeds germination, suggested that OsPII overexpression is likely to induce the expression of the cysteine proteinase, and secreted into endosperm to digest storage proteins as the nitrogen source for seedling development. According to these results, we concluded that the OsPII could enters into chloroplast and may play roles in nitrogen sensing or nitrogen metabolism regulation. |
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