臺灣博碩士論文加值系統

根菌促生菌 (Plant growth-promoting rhizobacteria, PGPR) 可藉由不同的機制影響植物生長及增加對生物及非生物性逆境抗性。本次研究中使用從土壤篩出的一株綠膿桿菌 (Pseudomonas Y1)及另一株芽孢桿菌 (Bacillus S4)，分別進行Pseudomonas Y1及Bacillus S4的蛋白水解酶活性及其代謝產物 (Y1-及 S4-M) 的抗氧化能力、螯鐵能力分析。並應用至重要的經濟作物蝴蝶蘭與香蕉苗測試Y1-M 與S4-M 對於植物生長及非生物性逆境的影響。由實驗結果分析，Y1-M 與S4-M 本身都具有良好的抗氧化能力而對蛋白水解酶活性及螯合二價鐵能力之影響為Pseudomonas Y1較高。在植物生長上，處理Y1-M及 S4-M 皆可增加兩種植物的葉綠素含量、總糖含量、抗氧化酵素活性，並提高植物體內木質素含量、二次代謝物質生合成，以促進植物生長。在非生物性逆境方面，處理Y1-M及 S4-M可藉由增加植物的抗氧化能力與二次代謝物質生合成，使植物細胞膜的穩定性增加，降低了細胞膜脂質過氧化的產物丙二醛 (malondialdehyde,MDA)、電子滲透率，以增加植物的對低溫、乾旱及鹽分逆境的耐受性，並加快了香蕉苗在乾旱逆境後新生葉片生長速度與光合作用效率。由香蕉的西方點墨法的結果可以得知，以Y1-M及 S4-M處理可增加植物逆境相關蛋白dehydrin和autophagy protein (ATG5) 以及抗氧化酵素superoxide dismutase (SOD)、catalase (CAT) 的蛋白累積量。這些結果顯示，Y1-M及 S4-M可增加植物抗氧化能力及二次代謝物質，使植物對非生物性逆境的耐受性提升。

Plant growth-promoting rhizobacteria (PGPR) had been reported to promote plant growth and improve plant resistant to biotic and abiotic stress through different kinds of mechanisms. In this study, strains of Pseudomonas (Y1) and Bacillus (S4) isolated from soil, and analyzed their protease activity, antioxidant activity and ion chelating activity of the metabolites (Y1-M and S4-M). We treated orchid and seedlings with Y1-M and S4-M to detect the effects on plant growth and response to abiotic stress. Our results showed that both Y1-M and S4-M exhibited antioxidant activity and Y1 exhibited better protease activity and ferrous ion chelating activity than S4. In term of plant growth test, both orchid and banana treated with Y1-M and S4-M increased chlorophyll content, total sugar content ,antioxidant enzyme activity, lignin content and induced plant secondary metabolism biosynthesis. As to tolerance to abiotic stress, orchid and banana treated with Y1-M and S4-M improved tolerance of cold stress, drought stress and salt stress through enhanced plants antioxidant activities and secondary metabolites biosynthesis. The results showed that treated plants improved cell membrane stability, reduced the accumulation of malondialdehyde (MDA), decreased electro leakage and enhanced the total new leaves numbers and photosynthesis efficiency of banana after recovering from drought stress. The western blot results of banana showed that treated plants increased stress-related proteins dehydrin, autophagy protein (ATG5) and antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) proteins accumulation, As a result, applied Y1-M and S4-M on plants could improve plant growth, enhance antioxidant activity, increase plant secondary metabolites biosynthesis and improve plants tolerance to abiotic stress.

目錄 1
圖次 3
中文摘要 4
英文摘要 5
壹、 前言 6
貳、 材料與實驗方法 14
實驗材料 14
一、 細菌genomic DNA 萃取 14
二、 細菌代謝成分抗氧化能力分析 15
三、 細菌蛋白水解酶活性分析 17
四、 Y1及S4代謝成分(Y1-M及S4-M)製備 17
五、 植物生長與生理分析 18
六、 植物生理試驗 18
七、 Western blot 22
八、 統計分析 23
參、 結果 24
一、 Pseudomonase Y1 及Bacillus S4 親源關係 24
二、 Pseudomonase Y1 及Bacillus S4 代謝物質分析 24
三、 Y1-M 及S4-M 對於蝴蝶蘭生長之影響 25
四、 Y1-M 及 S4-M 增加蝴蝶蘭對於低溫的耐受性 26
五、 Y1-M 及 S4-M 對於香蕉生長之影響 26
六、 Y1-M 與S4-M 增加香蕉對乾旱逆境的耐受性 27
七、 Y1-M 及S4-M 增加香蕉鹽分逆境的耐受性 28
八、 Y1-M 及S4-M 影響香蕉不同蛋白之表達 28
肆、 討論 30
伍、 參考文獻 34

圖次
圖1、Pseudomonase Y1 及 Bacillus S4 親源樹 44
圖2、Y1-M 及S4-M 代謝物質抗氧化能力比較 45
圖3、Y1-PH 及S4-PH 蛋白酶活性比較 46
圖4、Y1-M 及S4-M 對於蝴蝶蘭生長及氧化逆境影響 47
圖5、Y1-M 及S4-M 對於蝴蝶蘭二次代謝物質及木質素生合成之影響 48
圖6、Y1-M 及S4-M 增加蝴蝶蘭對於低溫逆境的耐受性 49
圖7、Y1-M 及S4-M 對於香蕉生長影響 50
圖8、Y1-M 及S4-M 對於香蕉生長及氧化逆境影響 51
圖9、Y1-M 及S4-M 對於香蕉二次代謝物質及木質素生合成之影響 52
圖10、Y1-M 及S4-M 對於香蕉乾旱逆境之影響 53
圖11、Y1-M 及S4-M 增加香蕉乾旱逆境後復水回復速度 54
圖12、Y1-M 及S4-M 增加香蕉乾旱逆境的耐受性 55
圖13、Y1-M 及S4-M 增加香蕉鹽分逆境的耐受性 56
圖14、Y1-M及S4-M 處理香蕉影響不同蛋白之表達量 57

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