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研究生: 林國英
Kuo-Yin Lin
論文名稱: 不同光質對甘藷小苗光合作用效率、碳水化合物代謝及抗氧化能力之影響
The effects of different light qualities on the photosynthesis efficiency, carbohydrate metabolism and antioxidant capacity of the sweet potato seedings
指導教授: 王玉麒
Wang, Yu-Chie
學位類別: 碩士
Master
系所名稱: 生命科學系
Department of Life Science
論文出版年: 2011
畢業學年度: 99
語文別: 中文
論文頁數: 98
中文關鍵詞: 光質發光二極體甘藷小苗光合作用效率碳水化合物代謝抗氧化能力
英文關鍵詞: light quality, light emitting diode (LED), sweet potato seeding, photosynthesis efficiency, carbohydrate metabolism, antioxidant capacity
論文種類: 學術論文
相關次數: 點閱:199下載:18
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  • 甘藷(Ipomoea batatas (L.) Lam.)葉是重要的抗氧化蔬菜,發光二極體(Light Emiting Diode, LED)則由於可改善傳統燈具壽命短、發熱大和發光效率差等缺點,並提供包括光質、光量、給光頻率及工作比等特定給光條件,成為近年來積極開發之新興人工光源。本論文利用LED提供甘藷小苗藍光(470 nm)與紅光(660 nm),除了探討抗氧化特性外,亦分析光合效率與碳水化合物的代謝受不同光質的影響情形,探討不同光質對甘藷小苗生理之影響。
    透過葉綠素螢光數據,經藍光、紅光或全光5天照射處理後,皆不至於對甘藷小苗光合系統Ⅱ造成破壞。由於紅光組甘藷小苗葉片之NPQ明顯低於藍光或全光處理,根據前人指出NPQ與zeaxanthin具正相關性,推測紅光會降低甘藷葉生合成zeaxanthin。
    抗氧化能力上,經藍光、紅光和全光處理後之甘藷小苗葉片在glutathion還原態(GSH)及氧化態(GSSG)的含量、ascorbate peroxidases(APX)活性和superoxide dismutase(SOD)活性上並未呈現顯著差異,但藍光處理之葉片表現較高的catalase(CAT)活性,且經藍光或紅光處理後之葉片H2O2含量均較全光低,推測藍光組可透過CAT降低活氧化物的累積。
    經測定可溶性醣類含量、澱粉含量與醣類代謝相關酵素活性,顯示不同光質雖未在澱粉含量造成影響,但藍光處理之葉片內葡萄糖、果糖及蔗糖含量皆明顯高於紅光處理,推測與藍光組較紅光組有較高之sucrose phosphate synthase(SPS)和invertase(INV)活性有關。此外全光處理之葉片具較高之acid INV和alkaline INV活性,紅光處理的acid INV和alkaline INV活性則均較藍光處理為低。不同光質下醣類代謝相關酵素活性以SPS和INV表現出顯著差異,但Real-time PCR偵測各酵素基因表現強度並無差別,推測此二者活性應受後轉譯調節所影響。不同光質5天照射雖未對葉片氣孔密度造成影響,但以藍光組氣孔開啟比例最高。
    總體而言,藍光處理之甘藷小苗生長良好、有較高之抗氧化能力,葉片內葡萄糖、果糖及蔗糖含量均較高,也具有較高的氣孔開啟比例,顯示在甘藷小苗生長上,藍光較全光、紅光為佳。

    The leaf of sweet potato (Ipomoea batatas (L.) Lam.) is an important antioxidant vegetable. The light emitting diode (LED) is a promising artificial light source because of its small mass and volume, and can provide specific light quality, light quantity, light frequency and duty ratio. This paper uses the LED as an artificial light source to provide blue light (470 nm) and red light (660 nm) for sweet potato seedings, and discusses the antioxidant capacity, photosynthesis efficiency, and carbohydrate metabilism of the sweet potato seedings cultivated under different light qualities.
    The data of chlorophyll fluorescence showed that photosystemⅡof the sweet potato seedings cultivated under blue, red or full-light for 5 days was not destroyed. NPQ of the sweet potato leaves cultivated under red light was significantly lower than that under blue or full-light. According to the previous research data, we can suggest that the leaves of sweet potato under red light might reduce the biosynthesis of zeaxanthin.
    The reduced glutathion (GSH) content, oxidized glutathion (GSSG) content, ascorbate peroxidase (APX) activity and superoxide dismutase (SOD) activity of the sweet potato leaves cultivated under blue, red or full-light for 5 days didn’t show the significant difference, but catalase (CAT) content of the sweet potato leaves cultivated under blue light was higher, and H2O2 content of the sweet potato leaves cultivated under blue or red-light was lower than that under full light. We can presume that the reactive oxygen species content of the sweet potato leaves cultivated under blue light reduced by CAT.
    The results of the contents of carbohydrates and the activities of the enzymes to participate in carbohydrate metabilism showed the different light qualities didn’t affect starch content of the sweet potato leaves, but glucose, fructose and sucrose contents of the sweet potato leaves cultivated under blue light were remarkably higher than those under red light. We can speculate that the activities of sucrose phosphate synthase (SPS) and invertase (INV) of the sweet potato leaves cultivated under blue light were higher than those under red light was the reason for foregoing results. Furthermore, the sweet potato leaves cultivated under full light had higher activities of acid INV and alkaline INV, and those under red light had lower activities of acid INV and alkaline INV than those under blue light. The activites of SPS and INV of the sweet potato leaves cultivated under blue, red, or full-light had significant difference, but the expressions of gene of SPS, sucrose synthase (Susy) , and INV displayed by Real-time PCR were not different. Therefore, the activity of SPS and INV might be affected by post-translation. Although the densities of the stroma of the sweet potato leaves cultivated under blue, red, or full-light didn’t have significant difference, the ratio of the open stroma of the sweet potato leaves cultivated under blue light was the highest.
    As a whole, the sweet potato seedings cultivated under blue light were growing abundantly, and the leaves of those had higher antioxidant capacity, more glucose, more fructose, more sucrose, and higher ratio of the open stroma. Those results indicate that blue light is better than red or full-light for sweet potato seedings.

    中文摘要----------------------------------------------------I 英文摘要--------------------------------------------------III 誌謝辭----------------------------------------------------VI 目錄----------------------------------------------------VIII 表目錄---------------------------------------------------XII 圖目錄--------------------------------------------------XIII 縮寫表----------------------------------------------------XV 第一章 緒論-----------------------------------------------01 1-1 甘藷的簡介-------------------------------------------01 1-2 文獻回顧---------------------------------------------02 1-2.1 不同光質對植物生長與生理之影響----------------------02 1-2.2 發光二極體的基本原理及在植物上的應用-----------------07 1-2.3 植物的光合作用效率---------------------------------09 1-2.3.1 葉綠素螢光原理---------------------------------10 1-2.3.2 葉綠素螢光與植物生理----------------------------12 1-2.4 植物的抗氧化反應-----------------------------------14 1-2.5 植物的碳水化合物代謝-------------------------------18 1-3 研究動機與目的---------------------------------------21 第二章 材料與方法------------------------------------------23 2-1 實驗材料與生長條件設定--------------------------------23 2-2 葉綠素螢光分析---------------------------------------24 2-3 抗氧化酵素活性分析------------------------------------26 2-3.1 Superoxide dismutase(SOD)之萃取與活性分析--------26 2-3.2 Catalase(CAT)之萃取與活性分析--------------------28 2-3.3 Ascorbate peroxidase(APX)之萃取與酵素活性分析-----30 2-3.4 GSH及GSSG含量分析---------------------------------31 2-4 H2O2含量分析-----------------------------------------32 2-5 碳水化合物成份分析------------------------------------34 2-5.1.1 可溶性醣的萃取---------------------------------34 2-5.1.2 醣類成份分析-----------------------------------36 2-5.2 澱粉含量分析--------------------------------------39 2-6 醣類代謝相關酵素佸性分析-------------------------------40 2-6.1 Intervase(INV)活性分析--------------------------40 2-6.2 Sucrose synthase(Susy)活性分析(分解方向)--------43 2-6.3 Sucrose phosphate synthase(SPS)活性分析---------46 2-6.3.1 丙酮粉末法-------------------------------------46 2-6.3.2 酵素活性分析-----------------------------------47 2-7 甘藷小苗之Real-time PCR分析--------------------------49 2-7.1 Total RNA的萃取----------------------------------49 2-7.2 反轉錄聚合連鎖反應(RT-PCR)----------------------51 2-7.3 即時定量聚合連鎖反應(Real-time PCR)--------------52 2-8 葉片上表皮氣孔變化------------------------------------54 第三章 結果-----------------------------------------------56 3-1 不同光質下甘藷小苗生長情形-----------------------------56 3-2 不同光質下甘藷小苗光合效率的比較------------------------56 3-3 不同光質對甘藷小苗葉片抗氧化能力之影響------------------56 3-4 不同光質對甘藷小苗葉片碳水化合物代謝之影響---------------57 3-5 不同光質對甘藷小苗葉片醣類代謝相關酵素活性與基因表現之影響-57 3-6 不同光質對甘藷小苗葉片氣孔密度與氣孔開啟比例之影響--------58 第四章 討論-----------------------------------------------72 4-1 不同光質下甘藷小苗生長情形-----------------------------72 4-2 不同光質下甘藷小苗光合效率的比較------------------------72 4-3 不同光質對甘藷小苗葉片抗氧化能力之影響------------------74 4-4 不同光質對甘藷小苗葉片碳水化合物代謝、相關酵素活性與基因表現 之影響----------------------------------------------75 4-5 不同光質對甘藷小苗葉片氣孔密度與氣孔開啟比例之影響--------80 第五章 結論-----------------------------------------------82 第六章 參考文獻--------------------------------------------84 表1-1. 葉綠素螢光相關參數對照表------------------------------13 表3-1. 比較不同光質下甘藷葉片之氣孔密度與氣孔開啟比例之差異-----71 圖1-1. 高等植物的蔗糖代謝-----------------------------------20 圖1-2. 高等植物碳水化合物代謝相關酵素於細胞內之分佈與交互作用----21 圖2-1. 生長箱內部配置與藍光(470 nm)、紅光(660 nm)光盤------24 圖3-1. 比較不同光質5天照射下甘藷小苗生長情形------------------59 圖3-2. 比較不同光質下甘藷小苗的光合效率差異-------------------60 圖3-3. 比較不同光質下甘藷葉片的Superoxide dismutase活性差異---61 圖3-4. 比較不同光質下甘藷葉片的Catalase活性差異---------------62 圖3-5. 比較不同光質下甘藷葉片的Ascorbate peroxidase活性差異---63 圖3-6. 比較不同光質下甘藷葉片的GSH及GSSG含量差異--------------64 圖3-7. 比較不同光質下甘藷葉片的H2O2含量----------------------65 圖3-8. 比較不同光質下甘藷葉片的碳水化合物含量------------------66 圖3-9. 比較不同光質下甘藷葉片的Sucrose phosphate synthase活性差 異-------------------------------------------------67 圖3-10.比較不同光質下甘藷葉片的Sucrose synthase活性差異-------68 圖3-11.比較不同光質下甘藷葉片的Invertase活性差異--------------69 圖3-12.比較不同光質下甘藷葉片各醣類代謝相關酵素基因表現差異-----70 圖3-13.甘藷葉片氣孔開啟情形---------------------------------71

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