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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王淑珍(Shu-Jen, Wang) | |
| dc.contributor.author | Chia-Wei Chang | en |
| dc.contributor.author | 張佳瑋 | zh_TW |
| dc.date.accessioned | 2021-06-16T08:14:40Z | - |
| dc.date.available | 2016-03-18 | |
| dc.date.copyright | 2014-03-18 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-02-13 | |
| dc.identifier.citation | 王揭晴 (2013) 光強度、氮素與通氣處理對水稻幼苗氮素吸收及同化之影響。國立台灣大學生物資源暨農學院系碩士論文。
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/58424 | - |
| dc.description.abstract | 碳水化合物的累積及運輸效能是決定作物產量的重要因素，目前已有研究指出低氮素或缺氮逆境會促進碳水化合物移往積儲組織及地下部。蔗糖為植物體中醣類運輸的主要形式，而蔗糖轉運蛋白 (sucrose transporter, SUT) 則為調控蔗糖進出韌皮部的主要蛋白質。本研究利用水稻 (Oryza sativa L. cv. Tainung 67) 懸浮細胞系統探討培養液中蔗糖、銨態氮及硝酸態氮含量改變對水稻蔗糖轉運蛋白基因家族成員OsSUT1、2、4及5表現之影響。研究結果顯示缺糖處理之下，細胞生長速度及細胞活力會極顯著下降，而OsSUT2、4及5的基因表現也隨之降低，但OsSUT1之基因表現則顯著誘導上升。在供糖處理之下，OsSUT2及4基因表現會受缺氮誘導，而此誘導現象於恢復供氮6小時後即消失，且銨態氮 (NH4+-N) 之效應較硝酸態氮 (NO3--N) 顯著。另一方面，OsSUT5之基因表現則受缺氮刺激所抑制，但於恢復供氮2天後可使OsSUT5基因表現回復，且NO3--N之效應較NH4+-N顯著。分析帶有不同OsSUT4啟動子長度片段接GUS報導基因之轉殖水稻懸浮細胞，發現OsSUT4啟動子上受缺氮調控之cis-acting elements位置可能位於OsSUT4轉譯起始點至上游434 bp間。 | zh_TW |
| dc.description.abstract | Accumulation and transport of carbohydrates are important for crop yield. Sucrose is the major sugar for long-distance translocation within plant tissue through phloem. Studies have suggested that nitrogen deficiency promotes translocation of non-structural carbohydrates to sink tissues in plants. Sucrose transporters (SUTs) play important roles in sucrose loading into phloem. In this study, effects of carbon and nitrogen nutrients on OsSUTs gene expressions in rice (Oryza sativa L.) suspension cells were investigated. The results showed that sucrose starvation significantly reduced cell growth rate and cell viability. Sucrose starvation also reduced OsSUT2, 4 and 5 expressions, but OsSUT1 gene expression was significantly induced in sucrose-starvated cells. In sucrose supplying condition, nitrogen starvation induced OsSUT2 and 4 gene expressions, and the effect was suppressed after 6 hours of nitrogen recovery treatment. The effect of nitrogen recovery treatment with ammonium (NH4+-N) on OsSUT2 and 4 gene expressions was more significant than that was treated with nitrate (NO3--N). On the other hand, OsSUT5 gene expression was inhibited under nitrogen starvation, but after nitrogen re-supplied for 2 days, OsSUT5 gene expression was increased to a level equivalent to control. Moreover, the NO3--N effect on OsSUT5 expression was more significant than NH4+-N. Based on OsSUT4 promoter activity analysis in transgenic rice suspension cells, it was suggested that the cis-acting elements related to nitrogen-starvation response were located at the region within 434 bp upstream of the translation start codon. | en |
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| dc.description.tableofcontents | 誌謝………………………………………………………………………………………i
中文摘要………………………………………………………………………………ii Abstract……………………………………………………………………………iii 目錄…………………………………………………………………………………v 表目錄…………………………………………………………………………………ix 圖目錄………………………………………………………………………………ix 附表及附圖目錄………………………………………………………………………x 縮寫字對照……………………………………………………………………………xi 前言…………….………………………………………………………………………1 1. 碳、氮兩大元素於植物體中之交互作用……………………………………1 2. 水稻對氮營養元素缺乏之反應及應對策略…………………………………2 3. 氮源變化對醣類分配之影響………………………………………………3 4. 醣類轉運蛋白於低氮逆境下扮演之角色……………………………………4 5. 水稻蔗糖轉運蛋白之功能及分類…………………………………………5 6. 碳、氮之訊息傳遞途徑……………………………………………………8 7. 與碳、氮調控相關之cis-acting element………………………………………9 8. 本論文之研究主題……………………………………………………………9 材料與方法…………………………………………………………………………11 1. 實驗材料……………………………………………………………………11 2. 水稻懸浮細胞系的建立……………………………………………………11 2.1 水稻癒傷組織 (callus) 的誘導………………………………………11 2.2 建立水稻懸浮細胞系…………………………………………………11 3. 材料處理……………………………………………………………………12 實驗一、水稻懸浮細胞於不同蔗糖濃度下進行缺氮處理…………………12 實驗二、水稻懸浮細胞於不同蔗糖濃度下缺氮後進行不同濃度及型態之短期恢復供氮處理………………………………………………12 實驗三、水稻懸浮細胞於缺氮後進行不同濃度及時間之恢復供氮處理.…………………………………………………………………12 實驗四、水稻懸浮細胞於缺氮後進行不同濃度及型態之長期恢復供氮處理.…………………………………………………………………13 實驗五、提升培養基醣類濃度處理.………………………………………13 實驗六、轉殖水稻懸浮細胞之缺氮處理…………………………………13 4. 細胞沉降體積 (sedimented cell volume) 測量……………………………13 5. 細胞活力檢.………………………………………………………14 5.1 Evans blue染色分析………………………………………………14 5.2 TTC染色分析…………………………………………………………14 6. 總可溶性醣類含量測定……………………………………………………14 7. 葡萄糖及蔗糖含量測定……………………………………………………15 7.1 可溶性醣類萃取………………………………………………………15 7.2 葡萄糖含量測定………………………………………………………15 7.3 蔗糖含量測定………………………………………………………16 8. 蛋白質含量測定………………………………………………………17 9. 銨態氮含量測定………………………………………………………17 10. 硝酸態氮含量測定………………………………………………………18 11. 基因表現分析……………………………………………………………………18 11.1 總RNA萃取…………………………………………………………………18 11.2 TURBO DNase 處理……………………………………………………19 11.3 RNA電泳…………………………………………………………………19 11.4 即時反轉錄聚合酶連鎖反應 (real-time RT-PCR)……………19 12. 轉殖水稻懸浮細胞GUS活性定量分析…………………………………20 12.1 蛋白質萃取………………………………………………………………………………20 12.2 蛋白質定量……………………………………………………………………………20 12.3 GUS活性測定…………………………………………………………………20 結果…………………………………………………………………………………………22 1. 不同蔗糖濃度培養下，缺氮處理對水稻懸浮細胞之生長、活力、碳氮含量、以及OsSUTs基因表現之影響…………………………………………22 1.1 不同蔗糖濃度培養下，缺氮處理對水稻懸浮細胞生長速率的影響………………………………………………………………………22 1.2 水稻懸浮細胞活力變化…………………………………………22 1.3 水稻懸浮細胞及培養基內之碳氮含量變化……………………………23 1.4 水稻懸浮細胞OsSUTs基因表現變化………………………………………25 2. 不同蔗糖濃度下，短期恢復供氮處理對水稻懸浮細胞OsSUTs基因表現之影響…………………………………………………………………………………………25 3. 長期恢復供氮處理對水稻懸浮細胞OsSUT5基因表現之影響……………26 3.1 水稻懸浮細胞OsSUT5基因表現受恢復供應混合氮之濃度與時間影響………………………………………………………………………………26 3.2 恢復供應單一型態氮對水稻懸浮細胞OsSUT5基因表現之影響………………………………………………………………………………26 4. 懸浮細胞內葡萄糖含量對OsSUTs基因表現之影響……………………………27 5. 不同蔗糖濃度培養下，缺氮處理對水稻懸浮細胞之OsRR6基因表現之影響……………………….....................………………………………………28 6. 缺氮處理對OsSUT4啟動子片段之調控……………………………………28 討論……………………………………………………………………………………29 1. 碳、氮營養源影響OsSUTs基因表現之可能調控機制………………………29 2. OsSUTs受醣類、氮、細胞分裂素、以及離層酸調控之啟動子區域探討………………………………………………………………………………………………………………………………30 3. OsSUTs於水稻懸浮細胞面臨缺氮時可能扮演之角色………………………32 4. OsSUTs於水稻幼苗面臨缺氮時可能扮演之角色……………………………33 5. 比較以水稻懸浮細胞系統或水稻幼苗探討基因調控之異同………………34 6. 結語與未來展望………………………………………………………………35 參考文獻……………………………………………………………………………………47 | |
| dc.language.iso | zh-TW | |
| dc.subject | 水稻 | zh_TW |
| dc.subject | 蔗糖轉運蛋白 | zh_TW |
| dc.subject | 懸浮細胞 | zh_TW |
| dc.subject | 缺氮 | zh_TW |
| dc.subject | 啟動子 | zh_TW |
| dc.subject | rice (Oryza sativa L.) | en |
| dc.subject | promoter | en |
| dc.subject | nitrogen starvation | en |
| dc.subject | sucrose transporter | en |
| dc.subject | suspension cells | en |
| dc.title | 碳/氮營養源對水稻懸浮細胞蔗糖轉運蛋白基因表現之調控 | zh_TW |
| dc.title | Regulation of Sucrose Transporter Gene Expressions by Carbon/Nitrogen Sources in Rice Cultured Cells | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 洪傳揚(Chwan-Yang, Hong),張孟基(Men-Chi, Chang),陳仁治(Jen-Chih, Chen),林淑怡(Shu-I, Lin) | |
| dc.subject.keyword | 水稻,蔗糖轉運蛋白,懸浮細胞,缺氮,啟動子, | zh_TW |
| dc.subject.keyword | rice (Oryza sativa L.),sucrose transporter,suspension cells,nitrogen starvation,promoter, | en |
| dc.relation.page | 66 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2014-02-13 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 農藝學研究所 | zh_TW |
| 顯示於系所單位： | 農藝學系 | |
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