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研究生:吳姿怡
研究生(外文):Tzu-I Wu
論文名稱:番茄WRKY31基因對非生物性逆境反應功能之研究
論文名稱(外文):Functional study of SlWRKY31 gene in response to abiotic stress
指導教授:潘怡君潘怡君引用關係
指導教授(外文):I-Chun Pan
口試委員:陳彥銘李嘉雯黃建誌
口試委員(外文):Yen-Ming ChenChia-Wen LiJian-Zhi Huang
口試日期:2019-07-06
學位類別:碩士
校院名稱:國立中興大學
系所名稱:園藝學系所
學門:農業科學學門
學類:園藝學類
論文種類:學術論文
論文出版年:2019
畢業學年度:107
語文別:中文
論文頁數:56
中文關鍵詞:番茄SlWRKY31轉錄因子非生物性逆境
外文關鍵詞:tomatoWRKYtranscription factorabiotic stress
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摘要………………………………………………………………………………i
Abstract…………………………………………………………………………..ii
目錄……………………………………………………………………………...iii
圖目次…………………………………………………………………………...vi
壹、前言……………………………………………………..……………………1
貳、前人研究…………………………………………………………………….2
一、WRKY轉錄因子……………………………………………………….2
二、植物逆境機制……………………………………………………………4
(一) 乾旱……………………………………………………………….4
(二) 鹽害……………………………………………………………….5
(三) 水楊酸 (Salicylic acid, SA) …………..………………………….6
(四) 茉莉酸 (Jasmonic acid, JA) ……………………………………...6
(五) 離層酸 (abscisic acid, ABA) …………..………………………...7
三、番茄WRKY基因...……………………………………………………...7
参、材料方法……………………………………………...……………………...9
1. 植物材料………………………………………………………………...9
2. SlWRKY31序列分析…………….…………………….………..………10
2. 1. 番茄組織各部分表現分析…………………...…………………10
2. 2. SlWRKY31蛋白質序列親緣比對………………………………..10
2. 3. 啟動子序列分析………………………………………………...10
3. 確認過量表現SlWRKY31之轉殖株內情形……......................……….11
3. 1. DNA萃取………………………………...……………………....11
3. 2. RNA萃取……………………………………………...…………11
3. 3. 轉殖株之聚合酵素鏈鎖反應 (Polymerase chain reaction, PCR) 分析……………………………………………………………..12
3. 4. 反轉錄PCR反應 (Reverse Transcription-PCR, RT-PCR)……12
3. 5. 即時聚合酵素連鎖反應(Real-time transcriptase polymerase chain reaction, RT- PCR)……………….……………………..….........13
4. SlWRKY31基因之表現模式……………………………………………13
5. 菸草轉植株逆境試驗…………………………………………….....…14
5. 1. 乾旱逆境處理……………………………………………...…....14
5. 2. 鹽害逆境處理……………………………………………...……14
5. 3. 含鹽培養基播種……………………………………………...…14
6. 逆境生理試驗測定..……………………………………………...……14
6. 1. 過氧化氫化學染色測定………………………………………...15
6. 2. 相對含水量測定…………………………………………….......15
6. 3. 離子滲漏率 (Electrolyte leakage, EL) 測定………………...…15
6. 4. 葉綠素含量測定…………………………………………….......16
6. 5. 丙二醛 (Malondialdehyde, MDA) 含量測定………………….16
6. 6. 脯胺酸 (Proline) 測定……………………..………….………..16
7. 統計分析…………………………………………………………..…...17
肆、結果……………………………………………………………………...…18
一、SlWRKY31分析………………………………………………………18
二、以PCR檢測菸草轉殖株內SlWRKY31基因及篩選基因HPTII…...…19
三、以RT-PCR檢測菸草轉殖株內SlWRKY31基因表現量……………19
四、乾旱逆境處理…………………………………………………………20
五、鹽害逆境處理…………………………………………………………21
伍、討論……………………………………………………………...…………24
陸、參考文獻……………………………………………………………...……43
柒、附錄………………………………………………..…………..….…...……52
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