臺灣博碩士論文加值系統

大豆蛋白水解物(Soybean protein hydrolysate, SPH)和海藻酸鈉(alginic acid, AlA)同屬於生物活性物(biostimulants)，研究結果顯示，這兩種生物活性物有促進植物生長和增強逆境下的耐受性。海藻酸鈉是從褐藻中分離的多醣體。前人研究指出，海藻酸鈉可以促進植物生長、提高作物產量，並做為防禦反應的激發子來對抗病原體感染，因此被作為新型的農業植物生長調解劑用於改善作物的品質。大豆蛋白水解物則利用Bacillus sp.分泌之蛋白酶生產的大豆蛋白水解物進行本次研究。本次研究利用微生物發酵製備SPH與AlA共同處理於目標植物，從結果顯示在阿拉伯芥低溫和缺水逆境處理下，SPH與AlA共同處理於滲透壓逆境時與單獨處理相較於對照組有增加下胚軸長度和側根數。SPH與AlA共同處理與單獨處理都有增加地下部根長與側根數相較於對照組於低溫逆境時。在番茄生長上，共同處理SPH與AlA與單獨處理下都有增加單一花絮上的開花數和降低電子測漏率 (Electrolyte leakage)相較於對照組，同時增加抗氧化酵素活性愈創木分過氧化物酶(GPX)，也會誘導CAT、AOX、LhcbI及PR3蛋白表現。在青江菜生長下共同處SPH與AlA可以抑制過氧化氫生成和MDA含量。由以上結果可知AlA結合SPH可以促進植物生長和抗氧化能力。

Protein hydrolysate (SPH) and alginic acid (AlA) are biostimulants that can promote plant growth, enhance plant tolerance to stressful environment. AIA is a polysaccharide isolated from brown algae. Previous studies showed that AIA could promot plant growth, increase crop yield, and act as an elicitor to trigger defense response against pathogen infection. In this study, a SPH prepared by microbial fermentation and AlA were used to treat target plants. Our results showed that Arabidopsis treated with SPH and AlA increased root length and lateral numbers under osmotic stress. Treatment of SPH and AIA can increase root length, lateral numbers and decreased electrolyte leakage under cold stress. Treatment with SPH and AlA also promoted Arabidopsis growth. Tomato plant treated with SPH and AlA exhibited increased number of flowers in a single inflorescence, and increased GPX antioxidant enzyme activity and increased protein accumulation included CAT, AOX , LhcbI and PR3. Brassica (Brassica rapa chinensis, variety Brassica) treated with SPH and AlA exhibited lower H2O2 accumulation and malonaldehyde (MDA) content. In summary, our results showed that SPH combined with AlA can promote plant growth and antioxidant enzyme activity.

目錄…………………………………………………………………………………………………………………………1
中文摘要-------------------------------------------------------------------------------------------------3
英文摘要-------------------------------------------------------------------------------------------------4
壹、 前言-------------------------------------------------------------------------------------------------5
貳、 材料與實驗方法--------------------------------------------------------------------------------13
1. 實驗材料-----------------------------------------------------------------------------------13
2. 實驗方法-----------------------------------------------------------------------------------13
一、 阿拉伯芥的培養及處理---------------------------------------------------------13
1. 清洗阿拉伯芥種子---------------------------------------------------------13
2. 植物生長試驗----------------------------------------------------------------13
3. 滲透壓逆境-------------------------------------------------------------------13
4. 低溫逆境處理----------------------------------------------------------------13
二、 大豆胜肽和海藻酸鈉處理番茄-----------------------------------------------14
三、 大豆胜肽和海藻酸鈉處理燕菜類作物-青江菜---------------------------14
四、 蛋白質相關技術-------------------------------------------------------------------14
1. 番茄蛋白質萃取------------------------------------------------------------14
2. Western blot 分析----------------------------------------------------------15
五、 生理分析相關技術----------------------------------------------------------------16
1. 過氧化氫含量之測定------------------------------------------------------16
2. Malonaldehyde (MDA)含量之測定-------------------------------------16
3. 抗氧化酵素活性------------------------------------------------------------16
4. 檢測電解質測漏率 (electrolyte leakage)----------------------------18
5. 統計分析----------------------------------------------------------------------18
參、 實驗結果------------------------------------------------------------------------------------------19
生物活性物對阿拉伯芥植株生長及抗氧化活性的影響---------------------------19
生物活性物對於葉菜類作物生長的影響-----------------------------------------------19
生物活性物降低青江菜氧化逆境---------------------------------------------------------19
生物活性物對於番茄生長與抗氧化活性的影響-------------------------------------20
生物活性物誘導番茄植株中蛋白的表現-----------------------------------------------20
生物活性物對阿拉伯芥低溫逆境的耐受性--------------------------------------------21
生物活性物對阿拉伯芥滲透壓逆境的耐受性-----------------------------------------21
肆、 討論------------------------------------------------------------------------------------------------22
伍、 參考文獻------------------------------------------------------------------------------------------33
圖次
圖1、大豆胜肽與alginate影響阿拉伯芥植株生長及抗氧化活性------------24
圖2、大豆胜肽與 alginate 對葉菜類植物生長的影響---------------------------25
圖3、大豆胜肽與alginate 對青江菜氧化逆境的影響----------------------------26
圖4、大豆胜肽與alginate對番茄開花時期的影響--------------------------------27
圖5、大豆胜肽與alginate 對番茄產量的影響--------------------------------------28
圖6、大豆胜肽與alginate 對番茄抗氧化活性的影響----------------------------29
圖7、大豆胜肽與 alginate誘導番茄蛋白的表現-----------------------------------30
圖8、大豆胜肽與 alginate 增加阿拉伯芥對低溫逆境的耐受性--------------31
圖9、大豆胜肽與 alginate 增加阿拉伯芥對滲透壓逆境的耐受性-----------32

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