應用自由基誘導物AAPH模擬氧化逆境: 研究大豆之抗氧化劑的生化防禦系統

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劉昱志(Yu-chi Liu) 查詢紙本館藏

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應用自由基誘導物AAPH模擬氧化逆境: 研究大豆之抗氧化劑的生化防禦系統
(Biochemical studies of antioxidative defense systems in soybean by a free radical initiator 2,2'-azobis(2-amidinopropane) hydrochloride)

相關論文

★ 應用自由基誘導物2,2’-Azobis (2-amidinopropane) hydrochloride (AAPH)模擬氧化逆境：研究大豆之抗氧化酵素Superoxide dismutase (SOD)的基因表現	★ 三氯乙烯及四氯乙烯對中國倉鼠卵巢細胞的毒性研究
★ 甘藷桃園二號對鹽分逆境之抗氧化物質的生化反應研究

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摘要(中)

環境的變化對植物的生長與發育有很深的影響。為了減低環境逆境的影響，植物發展出許多清除自由基的防禦機制，其中包括本研究室著重的ascorbate-glutathione循環。本研究是利用自由基誘導劑2,2’-azobis(2-amidinopropane) hydrochloride (AAPH)來誘發大豆 (Glycine max cv. Essex)之氧化逆境 (oxidative stress)，並研究其生理及生化上的反應。從本實驗結果中發現，在酵素性抗氧化物質部分，有一部份酵素如superoxide dismutase、ascorbate peroxidase、catalase及guaiacol peroxidase，其酵素活性隨時間的增加有上升的趨勢；另一部份如dehydroascorbate reductase及glutathione reductase之酵素活性則有下降的趨勢。利用ascorbate peroxidase活性電泳來研究其同功?活性之變化，其結果與酵素活性實驗相類似，但是並未能夠很清楚的辨認同功?的作用。關於非酵素性抗氧化物質之部分，如total ascorbate及total glutathione (還原態＋氧化態)之含量隨著時間增加有上升的趨勢；然而，氧化態的ascorbate 及glutathione卻無明顯的增加。由此研究得知，將50 mM AAPH噴灑於大豆植株表面確實可以誘發植物一部份之抗氧化機制；故推測AAPH可能使用於模擬植物於逆境中生長。

摘要(英)

The growth and development of plants are strongly influenced by different environmental stresses that can be initiated by oxidative stress. To cope with these stresses, plants have could develop a series of defense mechanisms and detoxification systems including ascorbate-glutathione cycle. The ascorbate-glutathione cycle is one of the most important antioxidative defense systems being in discussed plants. In this study, we investigated the physiological and biochemical responses of soybean (Glycine max cv. Essex), which is an ozone-tolerance cultivar, under oxidative stress initiated by azo compounds, 2,2’-azobis(2-amidinopropane) hydrochloride (AAPH). Results showed that 50 mM AAPH could cause visible injury on leaf tissues in soybean, reduced total chlorophyll contents and increased the levels of lipid peroxidation. Results also showed a slightly increase in activities of enzymes involved in detoxifying oxidative stress; such as ascorbate peroxidase (APX), superoxide dismutase, catalase and guaiacol peroxidase during the different incubation time. However, the activities of dehydroascorbate reductase and glutathione reductase were decreased. The APX activity in gel electrophoresis studies confirmed the enzyme activity data, but the activities of isozymes were not clearly identified. The levels of total ascorbate and glutathione contents were enhanced in soybean leaves when treated with 50 mM AAPH. However, the oxidized forms of ascorbate and glutathione contents were not significantly increased. This study suggests that some antioxidants involved in ascorbate-glutathione cycle in soybean can be induced by spraying 50 mM AAPH, and this technique could be used to screen the sensitivity of plants to environmental stress.

關鍵字(中)

★ 脂質過氧化反應
★ AAPH
★ 抗壞血酸-麩胱咁汰循環
★ 大豆
★ 氧化逆境

關鍵字(英)

★ 2'-azobis(2-amidinopropane) hydrochloride
★ ascorbate-glutathione cycle
★ soybean
★ oxidative stress
★ lipid peroxidation

論文目次

中文摘要…………………………………………………………1
英文摘要…………………………………………………………2
壹、前言…………………………………………………………3
貳、材料與方法..………………………………………………11
參、結果………………………………………………………22
肆、討論………………………………………………………26
伍、參考文獻…………………………………………………29
陸、圖…………………………………………………………37
圖一、葉綠體內超氧之形成及ascorbate-glutathione循環………37
圖二、大豆在植物生長箱之生長情形……………………………38
圖三、大豆處理50 mM AAPH及二次水後72小時至120小時之比較圖……39
圖四、大豆處理50 mM AAPH後total chlorophyll含量變化圖…40
圖五、大豆處理50 mM AAPH後MDA含量變化圖……………41
圖六、大豆處理50 mM AAPH後soluble protein含量變化圖…42
圖七、大豆處理50 mM AAPH後superoxide dismutase活性變化圖……43
圖八、大豆處理50 mM AAPH後ascorbate peroxidase活性變化圖……44
圖九、大豆處理50 mM AAPH後dehydroascorbate reductase活性變化圖 …………45
圖十、大豆處理50 mM AAPH後glutathione reductase活性變化圖……46
圖十一、大豆處理50 mM AAPH後catalase活性變化圖…………47
圖十二、大豆處理50 mM AAPH後guaiacol peroxidase活性變化圖……48
圖十三、大豆處理50 mM AAPH後total ascorbate含量變化圖…49
圖十四、大豆處理50 mM AAPH後還原態與氧化態ascorbate之比較圖…50
圖十五、大豆處理50 mM AAPH後total glutathione含量變化圖……51
圖十六、大豆處理50 mM AAPH後還原態與氧化態glutathione之比較圖…………52
圖十七、大豆處理50 mM AAPH後之APX活性電泳圖…………53

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指導教授

李賢淇

審核日期

2000-7-14

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