茉莉花油抑制葡萄銹病效果及有效成分之分析

Inhibitory effects of jasmine oil on grape rust and analysis of the active components

孫佩君

葡萄銹病 ； 葡萄銹病 ； 乳化植物油 ； 茉莉花油 ； 有效成分 ； 化學結構鑑定 ； grape rust ； Phakopsora ampelopsidis ； emulsified vegetable oil ； jasmine oil ； active components ； chemical structure analysis

屏東科技大學植物醫學系所學位論文

2016年

碩士

陳麗鈴

繁體中文

摘要 本研究以葉片圓盤法測試包括芝麻油等27種植物油對葡萄銹病之抑制效果。測試的油類以Tween 20乳化後加水稀釋200倍 (最終濃度4500 ppm)。葡萄葉片以直徑1.5 cm的打洞器製成圓盤後以乳化油處理3次，隨即接種，對照組以無菌水處理。每圓盤接種20 μl P. ampelopsidis夏孢子懸浮液 (105 spores/ml 0.1% Tween 20)，7-10天後觀察發病情形。結果顯示乳化茉莉花油抑制葡萄銹病的效果最好，發病率為0 %，對照組發病率為100%。孢子發芽試驗顯示乳化茉莉花油可抑制P. ampelopsidis夏孢子發芽。將其稀釋至500倍 (1800 ppm) 仍可降低50 %以上之罹病度。探討處理次數對防治效果之影響，結果顯示處理乳化茉莉花油2次，即能顯著降低病害的發病率及發病度。殘效測試顯示處理乳化茉莉花油後1天接種，仍可完全抑制病害發生。將茉莉花油通過矽膠管柱層析純化，獲得J1、J2、J3三個部份。孢子發芽測試顯示J1對P. ampelopsidis夏孢子發芽抑制率達99 %以上。將J1再次進行管柱層析，獲得J1-1、J1-2、J1-3。孢子發芽測試結果顯示J1-1 (200 ppm)及J1-2 (200 ppm) 可分別降低孢子發芽率99％及82％以上。透過質譜儀 (MS)以及核磁共振光譜儀 (NMR) 做化學結構分析，將J1-1及J1-2分別鑑定為二甲苯麝香(musk xylene) 和葵子麝香 (musk ambrette)，透過化學分析及生物檢測，我們鑑定出本研究所用之茉莉花油抑制銹病的有效成份，此為首次有研究顯示此二化合物可抑制銹病。

Abstract Twenty-seven vegetable oils were tested using a leaf disc method developed in this research for control of grape rust. Vegetable oils were emulsified by Tween 20 and diluted 200x (final concentration 4500 ppm) with water. The leaf discs (1.5 cm diam.) made from susceptible Kyoho grape leaves were treated with emulsified oil or water (control) 3 times before being inoculated with 20 μl rust spore suspension (105 spores/ml 0.1% Tween20), and placed in moisture chamber at 24℃. Infection rate was determined 7- 10 days after inoculation. The results showed that jasmine oil was the best in suppressing grape rust. Infection rate was 0% for leaf discs treated with jasmine oil and 100% for leaf discs treated with water. Jasmine oil significantly inhibited urediniospore germination of Phakopsora ampelopsidis. Jasmine oil diluted 500x (1800 ppm) still reduced the disease severity by 50%. Two applications of jasmine oil significantly reduced disease infection rate and disease severity. One day after treatment jasmine oil still completely suppressed the disease of grape rust. Fractination of Jasmine oil by silica gel column chromatography using different ratios of n-hexane and acetone yielded J1、J2、J3 fractions. The results of germination test showed that J1 fraction inhibited over 99% germination of P. ampelopsidis urediniospores. Furter fractionation of J1 by silica gel column chromatography, produced J1-1 、J1-2、J1-3 fractions. The results of germination test showed that compound J1-1 (200 ppm) and J1-2 (200 ppm) inhibited more than 99% and 82%, respectively, spore germination. The active fractions J1-1 and J1-2 were further analyzed by mass spectrometer (MS) and nuclear magnetic resonance spectrometer (NMR). J1-1 and J1-2 were chemically identified as musk xylene and musk ambrette respectively. In this research, the active ingredients of jasmine oil that inhibited grape rust were identified . This study is the first to show that these two compounds are inhibitory to grape rust.

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