阿拉伯芥hs49與78hs突變株之生理定性及其耐熱基因定位

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林育如(Yu-ru Lin) 查詢紙本館藏

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論文名稱

阿拉伯芥hs49與78hs突變株之生理定性及其耐熱基因定位
(Identification, Characterization and Gene Mapping of Arabidopsis thaliana hs49 and 78hs Mutants)

相關論文

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摘要(中)

植物因無法自由的移動，所以時刻都有可能面臨不適合生存的環境逆境。高溫逆境為主要的環境逆境之一。高溫會影響植物的生長及其產量。植物為了抵抗高溫逆境，必需演化出適當的保護機制。為了瞭解植物所演化出抵禦高溫逆境的機制，以及調控這些機制的遺傳因子，我們以前向式遺傳研究法(forward genetics approach)，利用化學突變劑甲基磺酸乙酯(ethylmethane sulfonate, EMS)篩選出兩株突變植株。其一為對長時間高溫敏感之突變株，命名為heat sensitive 49 (hs49)。其二為可耐受短時間熱休克逆境之突變株，命名為78hs (78 heat shock)。實驗結果顯示，hs49突變位址，是在阿拉伯芥AGI map 第五條染色體上，25,860 kb - 25,966 kb的區間範圍內。78hs突變位址，則是在阿拉伯芥編號At5g40280基因的第二個intron上，第134個鹼基由A變成T的突變點。此突變點經序列分析後，發現其為RNA splice辨認之處。過去有關At5g40280的研究指出，此基因突變或表現量降低時，會使植物對ABA具有過度敏感的性狀，且比野生種植株更能耐受乾旱逆境。生理實驗發現的結果也顯示出，78hs確實對ABA過度敏感，對乾旱逆境也有更高的耐受能力。然而，At5g40280如何保護植物免於高溫傷害之機制，仍然未知。

摘要(英)

Plants, being immobile and can not escape from their growing environments, are vulnerable to various environmental stresses and must develop effective protecting mechanisms for survival. One of the most typical environmental stresses plants encounter is high temperature stress. Heat temperature can affect plant growth and development. In aggriulature, high temperature can affect crop yield. In order to identify the genetic determinant that are essential for plant to cope with high temperature stress, hence better improve heat tolerance in crop, we have used a forward genetics approach to screen for mutants of Arabidopsis that have altered responses to high temperatures. Two mutants were isolated after ethyl-methane sulfonate (EMS)-mutagenesis. While hs49 (heat sensitive 49) is isolated because of its inability to tolerate sustained high temperature stress, 78hs (78 heat shock) is isolated because of its ability to tolerate heat shock stress. To identify the mutated loci of hs49 and 78hs, map-based cloning procedures was conducted. Results indicated that hs49 is located at a region between AGI map 25,860 kb-25,966 kb of the chromosome Ⅴ. The 78hs locus is mapped to At5g40280 gene. The mutation of 78hs is a single nucleotide change from A to T at the RNA splicing recongnized site of the 2nd intron. Previous studies have shown that At5g40280 null or down-regulrated plants are hypersensitive to ABA and can tolerate drought stress compare to those of wild-type. Physiological experiments showed that 78hs exhibits those previously described phenotypes. Nevertheless, the mechanisms by which 78HS contributes to protect plants against high temperature injuries are still unknown.

關鍵字(中)

★ 阿拉伯芥
★ 基因定位
★ 熱休克
★ 巴拉刈
★ 離層酸

關鍵字(英)

論文目次

中文摘要 I
英文摘要 II
致謝 III
總目錄 IV
圖目錄 VI
表目錄 VII
緒論 1
材料與方法 5
1. 突變株顯隱性之鑑定 5
2. 突變株之位址定位 6
3. 突變株之核苷酸定序 10
4. cDNA製備 11
5. 光照條件對hs49植物生長及其耐熱性之測試 13
6. 巴拉刈(paraquat, MV)對hs49生長表現型之測試 13
7. hs49和78hs幼苗對抗離層酸(abscisic acid, ABA)逆境之測試 14
8. 鹽分逆境對hs49與78hs生長之測試 14
9. 熱休克處理(heat shock)對78hs表現型之測試 14
10. 78hs耐受熱休克性狀與不耐受長時間熱處理性狀之驗證 14
11. 78hs成株植物晚開花現象之觀察 15
12. 78hs成株植物耐乾旱現象之觀察 15
結果 16
1. hs49突變株對於持續性高溫處理之表現型 16
2. hs49突變株之顯、隱性鑑定 16
3. hs49之位址定位 16
4. 光照條件對hs49植物生長及其耐熱性之測試 17
5. 巴拉刈(paraquat, MV)對hs49生長表現型之測試 18
6. hs49幼苗對抗離層酸 (abscisic acid, ABA)逆境之測試 18
7. 鹽分逆境對hs49之測試 19
8. 78hs突變株之表現型 19
9. 78hs突變株之顯、隱性鑑定 20
10. 78hs之位址定位 20
11. 78hs 突變株之核苷酸定序 21
12. 78hs RNA splicing3’端辨認剪切位置之確認 22
13. 78hs耐受熱休克性狀與不耐受長時間熱處理性狀之驗證 22
14. 78hs成株植物晚開花之測試 23
15. 78hs幼苗對抗離層酸(abscisic acid, ABA)逆境之測試 23
16. 78hs成株植物耐乾旱之測試 24
17. 鹽分逆境對78hs之測試 24
討論 25
1. hs49突變基因定位及生理實驗之探討 25
2. 78hs基因功能之探討 25
3. 總結 29
文獻參考 31

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指導教授

吳少傑

審核日期

2014-10-28

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