博碩士論文 972204007 詳細資訊




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姓名 劉依欣(Yi-Hsin Liu)  查詢紙本館藏   畢業系所 生命科學系
論文名稱 水稻第一族小分子量熱休克蛋白質OsHSP16.9A及OsHSP18.0之生理功能分析
(Physiological function assay of class I small heat shock proteins, OsHSP16.9A and OsHSP18.0)
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摘要(中) 近年來全球暖化對於作物產量造成威脅,因此如何幫助水稻抵抗惡劣環境及提升作物產量皆現在重要的議題。水稻是世界上重要的糧食作物,其供應全球一半以上的人口食用,除此之外它亦是研究單子葉植物的模式植物,可提供我們研究其基因功能。根據我們先前的研究,除熱誘導外,水稻第一族小分子熱休克蛋白質中,OsHSP16.9A會在種子成熟時期表現及OsHSP18.0會受到重金屬銅或鎘誘導表現。為了更進一步了解它們的功能,我們建立OsHSP16.9A和OsHSP18.0持續性表現的轉殖株 (OsHSP16.9A-OE, OsHSP18.0-OE),並建立抑制OsHSP16.9A、OsHSP18.0和sHSPs表現的轉殖株(Oshsp16.9A-RNAi, Oshsp18.0-RNAi, Osshsp-RNAi)。試驗結果發現OsHSP16.9A-OE種子和幼苗與OsHSP18.0-OE的幼苗皆具有對高溫的耐受性;Oshsp18.0-RNAi及Osshsp-RNAi不論在種子或幼苗對於高溫是敏感的。而利用基因表現或缺失的方式分析sHSPs是否參與在水稻的生長與發育中,結果發現Osshsp-RNAi植株其分櫱數會增加同時生長也較遲緩,因此從這個結果,我們推論sHSPs不只扮演著chaperone的功用保護失去活性的蛋白質外,同樣也參與在水稻的生長過程中。此外,我們也發現在Cu及Cd處理下,OsHSP18.0-OE植株比OsHSP16.9A-OE 和野生型植株對於重金屬逆境有較高的耐受性,而所受的氧化性傷害也是較少的,另OsHSP18.0-OE會降低銅誘導的細胞膜破損。這些結果都說明了OsHSP18.0-OE在水稻對抗重金屬逆境時扮演著重要的角色。
摘要(英) Recently global warming is threatening the crop yield. It becomes a critical issue to improve tolerance of rice to harsh environment as well as crop yield. Rice (Oryza sativa) is one of the most important crops in the world and feeds nearly 50% of the world’s population. Besides, it can be a model plant for providing insights into gene functions of monocotyledonous plants. Based on our previous data, we found that rice class I small heat shock proteins (sHSP-CIs) selectively expressed during seed maturation (OsHSP16.9A) and Cu or Cd treatment (OsHSP18.0). To further characterize physiological function of sHSP-CIs, we constructed two transgenic rice plants constitutively overexpressing OsHSP16.9A (OsHSP16.9A-OE) and OsHSP18.0 (OsHSP18.0-OE) and three transgenic rice plants repressing OsHSP16.9A (Oshsp16.9A-RNAi), OsHSP18.0 (Oshsp18.0-RNAi), and sHSPs (Osshsp-RNAi). Our results indicated that Oshsp16.9A-OE seeds and seedlings and Oshsp18.0-OE seedlings had higher thermotolerance than wild type during heat stress. Besides, Osshsp-RNAi plants were sensitive to heat stress. Interesting, gain- and loss-of-function analyses identified that sHSPs involved in rice growth and development. Osshsp-RNAi plants were shown increased in tiller number and delayed in growth. These results suggest that sHSPs can not only function as chaperone to protect denatured proteins but also involve in rice growth process. Furthermore, OsHSP18.0-OE plants showed more resistance than OsHSP16.9A-OE and wild-type plants under Cu and Cd treatment. Similarly, OsHSP18.0-OE plants were less oxidative damage. In addition, OsHSP18.0-OE plants reduced Cu-induced membrane damage. These results indicated OsHSP18.0-OE play important role in heavy metal stress.
關鍵字(中) ★ 水稻
★ 溫度
★ 熱休克蛋白質
★ 重金屬
關鍵字(英) ★ temperature
★ heavy metal
★ heat shock protein
★ rice
論文目次 目錄
摘 要 i
Abstract ii
目錄 iii
縮寫對照表 ivii
壹、緒論 1
環境逆境 1
植物對抗逆境之反應 2
水稻基因體功能研究 2
貳、文獻探討 4
熱休克蛋白質 4
熱逆境反應 5
低溫逆境 6
重金屬逆境 7
作物產量 8
研究起源與目的 9
參、材料與方法 11
一、水稻農桿菌轉殖 11
二、基因表現分析 12
三、生理功能分析 15
四、水稻產量分析 20
肆、結果 23
Class I small heat shock protein 轉植株篩選與mRNA表現情形 23
轉殖株生長情形 24
轉殖株產量分析 24
水稻耐熱性轉植株篩選 26
轉植株逆境抗性分析 27
伍、討論 30
轉殖株生長高度及分糵數分析 30
植株花粉受熱後與產量的關係 31
耐熱性 32
轉殖株經重金屬逆境下抗氧化酵素表現情形 33
轉殖株經重金屬逆境下離子滲漏情形 35
未來研究方向 35
陸、參考文獻 37
柒、圖表 46
捌、附錄 70
圖目錄
圖一、轉殖株水稻中OsHSP16.9A及OsHSP18.0表現情形。 47
圖二、轉殖株水稻中Osshsp-RNAi表現情形。 48
圖三、OsHSP16.9A過量與抑制表現植物生長高度分析。 49
圖四、OsHSP18.0過量與抑制表現植物生長高度分析。 50
圖五、抑制sHSPs表現植物生長高度分析。 51
圖六、轉殖株生長分櫱數分析。 52
圖七、抽穗期各轉植株高度比較。 53
圖八、正常情況下,花粉存活情形。 54
圖九、熱處理後,花粉受熱後表現情形。 55
圖十、冷處理後,花粉遇冷後表現情形。 56
圖十一、冷處理後,植株抽穗情形。 57
圖十二、轉殖株經熱處理、冷處理後光合作用效率之差異。 58
圖十三、種子耐熱性分析。 59
圖十四、幼苗耐熱性分析。 60
圖十五、轉殖株經氯化鎘處理,葉綠素及抗氧化酵素之表現 61
圖十六、轉殖株經硫酸銅處理,葉綠素及抗氧化酵素之表現。 62
圖十七、轉殖株經氯化鎘處理,DAB染色情形及MDA含量測定。 63
圖十八、轉殖株經硫酸銅處理,DAB染色情形及MDA含量測定。 64
圖十九、轉殖株經硫酸銅處理,離子滲漏情形分析。 65
圖二十、野生型植株與轉殖株之種皮顏色。 66
Table1、野生型植株與轉殖株在正常生長條件下,分櫱數、稔實率、
穗長及種子數的統計分析。……………………………………………67
Table2、野生型植株與轉殖株在開花時期,利用38℃高溫處理三小時三天後生長情形,分櫱數、稔實率、穗長及種子數的統計分析。……………68
Table3、野生型植株與轉殖株在開花時期,利用6℃低溫處理24小時後生長情形,分櫱數、稔實率、穗長及種子數的統計析。…………................69
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指導教授 葉靖輝(Ching-Hui Yeh) 審核日期 2011-1-20
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