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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張英? | |
| dc.contributor.author | Cheng-En Lee | en |
| dc.contributor.author | 李承恩 | zh_TW |
| dc.date.accessioned | 2021-06-17T06:00:48Z | - |
| dc.date.available | 2021-02-14 | |
| dc.date.copyright | 2019-02-14 | |
| dc.date.issued | 2019 | |
| dc.date.submitted | 2019-02-12 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/71442 | - |
| dc.description.abstract | 在植物體中，有一類蛋白質因為胺基酸序列與動物體之離子型麩胺酸受體 (ionotropic glutamate receptors，iGluRs) 相似，因此命名為glutamate receptor-like proteins (GLRs)。在阿拉伯芥中，有20個基因組成麩胺酸受體家族，並且普遍被認為是植物細胞膜上之鈣離子通道，可調控植物許多生理反應的發生。先前研究證實，atglr3.7突變株在鹽逆境處理下之種子萌芽率會顯著低於野生株 (Col-0)。此外，14-3-3s為廣泛存在於真核生物之鷹架蛋白，通常以同質或異質二聚體的形式與磷酸化蛋白結合，並調控蛋白在細胞內的生理功能。實驗室先前研究發現，AtGLR3.7第860號的絲胺酸 (Ser-860)，可以被鈣離子依存性蛋白質激酶 (CDPK) 磷酸化，並且At14-3-3ω會在相同位置與AtGLR3.7結合。因此，本研究主要探討是否AtGLR3.7 Ser-860為一個重要的位點，以及在鹽逆境處理下AtGLR3.7之功能。實驗結果顯示，AtGLR3.7與At14-3-3ω會在菸草 (Nicotiana benthamiana) 下表皮細胞膜上有交互作用，且將AtGLR3.7第860號的絲胺酸突變成丙胺酸 (S860A) 之後，與14-3-3ω之交互作用因此消失；此外，AtGLR3.7大量表現株及AtGLR3.7-S860A大量表現點突變株在125 mM鹽逆境處理下之主根根長較不敏感。atglr3.7點突變株在鹽逆境處理下之種子萌芽率顯著被抑制。本研究進一步發現，glr3.7突變株在鹽逆境處理下，細胞質鈣離子濃度顯著低於野生株。綜合以上實驗結果，阿拉伯芥麩胺酸受體3.7第860號的絲胺酸參與鹽分反應。 | zh_TW |
| dc.description.abstract | In planta, glutamate receptor-like proteins (GLRs) are similar to animal ionotropic glutamate receptors (iGluRs). In Arabidopsis, there are 20 genes which encode GLRs family. GLRs are calcium channels on the cell membrane and can modulate several physiological function in the cells. Previous study indicated that under salt stress condition, seeds germination rate of atglr3.7 mutation line is significantly lower than Col-0. On the other hand, 14-3-3s are scaffold proteins which can bind to phosphorylated proteins with homo- or hetero- dimer form to modulate the physiological function of binding proteins in eukaryotes. Our previous studies indicated that calcium-depedent protein kinase (CDPK) can phosphorylate Ser-860 of AtGLR3.7. Furthermore, 14-3-3ω can bind to the same phosphorylation site of AtGLR3.7. Therefore, this study aimed to investigate whether Ser-860 of AtGLR3.7 is an important phosphorylation site and whether AtGLR3.7 is involved in the regulation of salt stress responses. The results showed that AtGLR3.7 could bind to At14-3-3ω on the plasma membrane of the lower epidermis of Nicotiana benthamiana. Furthermore, there were no physically interaction between AtGLR3.7 with Ser860 point mutation (AtGLR3.7- S860A) and At14-3-3ω. On the other hand, the primary root length of AtGLR3.7 overexpression lines and AtGLR3.7- S860A overexpression lines were more insensitive to 125 mM NaCl salt stress treatment. The seed germination rate of atglr3.7 mutation line was significantly inhibited under salt stress condition. Besides, cytosolic calcium concentration of atglr3.7 mutation line was significantly lower than Col-0 under salt stress condition. In summary, Ser-860 of glutamate receptor-like protein GLR3.7 is involved in salt response in Arabidopsis. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T06:00:48Z (GMT). No. of bitstreams: 1 ntu-108-R04b42018-1.pdf: 2664188 bytes, checksum: 1ee35e4e939625180571b7f0ef92abff (MD5) Previous issue date: 2019 | en |
| dc.description.tableofcontents | 誌謝 ii
摘要 iii Abstract iv 常用縮寫與全名對照表 v 目錄 vii 圖目錄 x 附表目錄 xi 附圖目錄 xii 第一章 前言 1 1.1 非生物性逆境 1 1.1.1鹽分逆境反應 1 1.1.2 離層酸abscisic acid (ABA) 2 1.2 鈣離子訊號傳遞 2 1.3 鈣離子通道 3 1.4 麩胺酸受體 (Glutamate receptor) 4 1.4.1 植物麩胺酸受體 (glutamate receptor-like proteins) 4 1.4.2 阿拉伯芥麩胺酸受體之表現 4 1.4.3 阿拉伯芥麩胺酸受體為鈣離子通道 5 1.4.4 阿拉伯芥麩胺酸受體之功能 6 1.5植物14-3-3蛋白質 6 1.6 研究動機 7 第二章 材料與方法 9 2.1 植物材料及生長條件 9 2.2 植物材料篩選及基因型檢測 9 2.2.1 轉殖株篩選 9 2.2.2 基因型檢測 9 2.3 細菌轉型作用 (tansformation) 10 2.4 植物RNA萃取及即時定量PCR分析 10 2.5 植物外表型檢測 11 2.5.1植物根部生長觀察 11 2.6 次細胞定位 (subcellular localization) 及雙分子螢光互補系統 [bimolecular fluorescence complementation (BiFC)] 11 2.6.1質體建構 11 2.6.2 觀察subcellular localization及BiFC分析 11 2.7 水母素冷光試驗 (Aequorin bioluminescence assay) 12 第三章 結果 13 3.1 T-DNA插入突變株atglr3.7之篩選 13 3.2 離層酸處理下相關標記基因RD29B及RAB18之表現 13 3.3 鹽處理下相關標記基因RD29A及RAB18之表現 13 3.4 AtGLR3.7之次細胞定位 14 3.5 GLR3.7與14-3-3ω之蛋白質交互作用 14 3.6 AtGLR3.7大量表現株幼苗在鹽逆境處理下之主根根長 15 3.7 Atglr3.7突變株在鹽逆境處理下之種子萌芽率 15 3.8 Atglr3.7突變株在離層酸處理下之種子萌芽率 16 3.9 在鹽逆境處理下之細胞質鈣離子濃度 16 第四章 討論 17 參考文獻 21 圖 32 附表 46 附圖 49 | |
| dc.language.iso | zh-TW | |
| dc.subject | 14-3-3ω | zh_TW |
| dc.subject | 磷酸化 | zh_TW |
| dc.subject | 根長 | zh_TW |
| dc.subject | 鹽逆境 | zh_TW |
| dc.subject | 鈣離子 | zh_TW |
| dc.subject | GLR3.7 | zh_TW |
| dc.subject | phosphorylation | en |
| dc.subject | 14-3-3ω | en |
| dc.subject | salt stress | en |
| dc.subject | root length | en |
| dc.subject | calcium | en |
| dc.subject | GLR3.7 | en |
| dc.title | 阿拉伯芥麩胺酸受體3.7第860號的絲胺酸參與鹽分反應 | zh_TW |
| dc.title | Glutamate receptor-like protein GLR3.7 Ser-860 is involved in salt response in Arabidopsis | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 107-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 葉國楨,梁國淦,靳宗洛,王雅筠 | |
| dc.subject.keyword | GLR3.7,14-3-3ω,鹽逆境,根長,鈣離子,磷酸化, | zh_TW |
| dc.subject.keyword | GLR3.7,14-3-3ω,salt stress,root length,calcium,phosphorylation, | en |
| dc.relation.page | 60 | |
| dc.identifier.doi | 10.6342/NTU201900466 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2019-02-12 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 植物科學研究所 | zh_TW |
| 顯示於系所單位： | 植物科學研究所 | |
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