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標題: 布袋蓮萃取物與乳化葵花油防治南方型玉米銹病的效果評估
Evaluation for efficacy of water hyacinth extract and emulsified sunflower oil on control of southern corn rust caused by Puccinia polysora
作者: 朱炯寰
Chu, Chiung-Hun
關鍵字: 南方型玉米銹病
Puccinia polysora
葵花油乳劑 (SFOC18605)
Eichhornia crassipes
sunflower oil
sunflower oil emulsion (SFOC18605)
particle size
出版社: 植物病理學系所
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摘要: 由Puccinia polysora引起的南方型玉米銹病 (southern corn rust) 在全球玉米種植區常造成重大經濟損失,雖然化學合成殺菌劑能有效防治南方型玉米銹病,但其對人類健康及環境的負面效應已引起高度關注,發展替代的防治方法遂成為當前急迫的課題。本文主要目的在於評估布袋蓮 (Eichhornia crassipes) 萃取物與乳化植物油防治南方型玉米銹病的效果。首先檢測布袋蓮萃取物之抑菌族譜,發現其可以顯著抑制P. polysora夏孢子的發芽百分率與發芽管的生長。進一步,將布袋蓮甲醇萃取物分別以乙酸乙酯 (Ethyl acetate)、正丁醇 (n-Butanol) 及水進行液相-液相分割萃取,獲得各溶劑萃取的濃縮成分,經抑菌試驗分析證實乙酸乙酯萃取物100 ppm具有抑制96.7%P. polysora夏孢子發芽的效果。隨後將乙酸乙酯萃取物透過正相管柱層析 (column chromatography, CC) 進行初步分離後,再利用HPLC正相管柱分別以不同比例之正己烷、乙酸乙酯及甲醇混合溶液作為沖提劑,依極性不同逐次流洗,總共獲得3個分離物(EA-06-03-01, -02, -03),其中EA-06-03-03在濃度1,000 ppm可完全抑制夏孢子的發芽,以1H-NMR進行結構分析鑑定其有效成分為4-Hydroxybenzaldehyde。此外,將6種植物油分別以高速攪拌方式製成乳化植物油後,用於防治南方型玉米銹病,結果顯示各植物油皆能降低P. polysora夏孢子之發芽管長度與減少夏孢子堆的形成,其中以葵花油的效果最佳,其濃度在1,000 ppm時能降低夏孢子發芽管長度達40.6%及減少66.7%夏孢子堆形成,且防治效果隨著濃度的增加而提升。自4種不同碳鏈長脂肪酸化合物篩選出碳鏈長18之脂肪酸化合物 (C18-FA),其濃度於250 ppm可使夏孢子發芽管畸形,500 ppm可減少夏孢子於玉米葉表氣孔形成附著器,5,000 ppm則可完全抑制夏孢子發芽。分析4種不同比例葵花油與C18-FA混合之葵花油乳劑 (sunflower oil emulsion) 對P. polysora夏孢子發芽的影響,結果發現60%葵花油與5% C18-FA調製成葵花油乳劑 (SFOC18605) 之100倍稀釋液可有效防治93-97.1%夏孢子堆於玉米葉表形成。油粒徑大小會影響葵花油乳劑抑制夏孢子發芽的功效,試驗結果發現葵花油乳劑之油粒徑為500 nm時,其100倍稀釋液可完全抑制P. polysora夏孢子發芽,但卻不具殺菌效果。在接種P. polysora夏孢子前24小時噴灑葵花油乳劑SFOC18605防治南方型玉米銹病的效果優於接種夏孢子後24小時的噴佈處理;此外,每週於玉米葉片噴佈葵花油乳劑一次,連續2次,其防治南方型玉米銹病的效果最佳。
The southern corn rust of maize (Zea mays L.) caused by Puccinia polysora is a destructive disease of major economic losses in almost maize production areas of the world. Although several fungicides have been found to give good control of the disease, high concern for adverse effects of pesticides on human health and the environment makes the development of alternative methods for disease management an urgent need. The purposes of the study were to evaluate the effect of Eichhornia crassipes (water hyacinth) extract and emulsified plant oils on control of P. polysora. The extracts of E. crassipes showed varied inhibitory effects on spore germination of plant fungal pathogens, however, they all showed significantly effectiveness in inhibiting urediniospore germination and germ tube elongation of P. polysora. The methanol-soluble extract of E. crassipes was concentrated under reducing pressure and further divided into ethyl acetate-, n-butanol- and water-soluble fractions by using liquid-liquid partition. All fractions were concentrated and evaluated their antifungal activity. At a concentration of 100 ppm, ethyl acetate-soluble extract showed significantly effectiveness in inhibiting 96.7% urediniospore germination of P. polysora. To identify the active components, the ethyl acetate-soluble extract was concentrated and subjected to column chromatography with silica gel using n-hexane, ethyl acetate and methanol as eluents with increasing polarity, and activity fraction then subjected to normal phase high performance liquid chromatography using hexane and ethyl acetate as eluents with increasing polarity. Three fractions (EA-06-03-01, -02, -03) were obtained and then tested individually for antifungal activity. The concentration of EA-06-03-03 at 1,000 ppm was found to completely inhibit urediniospore germination of P. polysora. Nuclear magnetic resonance (NMR) analysis of EA-06-03-03 was identified as 4-hydroxybenzaldehyde. In addition, the 6 kinds of plant oils were evenly emulsified by high speed homogenization for control of southern corn rust. Each of emulsified plant oil showed significantly effectiveness in inhibiting germ tube length of urediniospore and pustule formation of P. polysora. At a concentration of 1,000 ppm, sunflower oil was significantly effective in inhibiting 40.5% germ tube length of urediniospores and 66.7% pustule formation of P. polysora. The effectiveness of sunflower oil was increased with increase in concentration. 18-carbon fatty acid compound (C18-FA) was selected from four different carbon chain length of fatty acid compounds by using antifungal activity tests. The C18-FA was able to cause germ tube deformed at 250 ppm, reduce percentage of appressorial formation on stomata of corn leaf surface at 500 ppm, and completely inhibit urediniospore germination at 5,000 ppm. Evaluation for efficacy of 4 kinds of sunflower oil emulsions obtained from mixing with different ratios of C18-FA and sunflower oil on urediniospore germination of P. polysora was conducted. Sunflower oil emulsion (SFOC18605), a mixture of C18-FA and sunflower oil by 5 : 60 ratio (w/v), at a concentration of 100-fold dilution was significantly effective in inhibiting 93-97.1% pustule formation of P. polysora on corn leaf surface. The particle size of sunflower oil emulsion could influence its efficacy on reducing urediniospore germination. Sunflower oil emulsion (SFOC18605) with the particle size of 500 nm at a concentration of 100-fold dilution showed complete inhibition of urediniospore germination of P. polysora, but it didn’t present fungicidal effect. Spraying sunflower oil emulsion (SFOC18605) on corn leaf surface one day prior to inoculation of urediniospores showed the better efficacy on control of southern corn rust than spraying it one day after inoculation. Furthermore, weekly spraying sunflower oil emulsion (SFOC18605) on corn leaf surface for 2 times showed the best efficacy on control of southern corn rust.
其他識別: U0005-2208201300273200
Appears in Collections:植物病理學系



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