- 學位論文
五種精油與其主要成分對菜豆立枯病原菌（立枯絲核菌與茄鐮孢菌）的防治效果
Efficacy of Five Essential Oils and Their Major Compounds against Damping-off Pathogens (Rhizoctonia solani and Fusarium solani) on Phaseolus vulgaris
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摘要
立枯絲核菌（Rhizoctonia solani）與茄鐮孢菌（Fusarium solani）為常見的病原菌，它們能造成立枯病的發生並嚴重減少作物的產量。研究指出，許多植物的所含成分能減少立枯病的發生與其嚴重性。本研究測試植物精油（肉桂、丁香、檸檬草、山胡椒、澳洲尤加利）與其主要成分在試管內與活體實驗中，對這兩種病原菌的抗真菌作用，以期能制定出對非目標生物毒性較小、殘留量較少的病原菌防治對策。在試管內實驗中，PDA培養基和四種濃度精油（50、100、200或400 µg/mL）混合，以觀察其抗真菌活性。結果顯示，肉桂精油的抗真菌效果最佳，在100 µg/mL與400 µg/mL的濃度下能對病原菌的生長造成100% 抑制效果。其次為檸檬草精油可抑制立枯絲核菌，丁香精油能抑制茄鐮孢菌。澳洲尤加利精油則無法有效抑制立枯絲核菌和茄鐮孢菌。氣相層析質譜儀分析結果顯示，肉桂精油含有74.04%桂皮醛、丁香精油含84.56%丁香油酚、檸檬草精油含36.72%香葉醛與28.81%橙花醛。這些主成分可能與精油對病原菌的高抑制效果有關。在試管內實驗中，50 µg/mL桂皮醛能有效抑制立枯絲核菌與茄鐮孢菌的菌絲生長。在溫室環境下，對菜豆種子分別施以0.1%或1%的肉桂、丁香、檸檬草精油或其主要成分，並對受2%立枯絲核菌感染之土壤施以2%精油或其主成分。無菌基質被2%（w/w）的立枯絲核菌接種原感染。結果顯示，0.1%與1%的肉桂精油能有效防止菜豆種子被病原菌感染，而種子萌芽後的疾病發生率分別為24%和46%，疾病嚴重程度為24%和38%，存活率為40%和60%。肉桂精油對土壤的抑菌效果比對種子更佳，它能降低疾病發生率與嚴重程度，並能提升菜豆的農藝性狀本。研究結果證實，肉桂、檸檬草與丁香精油和其主成分皆能對立枯絲核菌與茄鐮孢菌展現出良好的抗真菌活性，然而僅有肉桂精油能在土壤環境中產生抑菌效果。由此可知，肉桂精油具有防治植物立枯病的潛力，並能取代化學農藥，避免對人體健康與自然環境造成危害。
並列摘要
Rhizoctonia solani and Fusarium solani are very common phytopathogens around the world that causes one of the most damaging disease “Damping off” resulting in significant crop losses. Use of natural compound derived from many plants have been reported to reduce disease incidence as well as disease severity in many varieties of crops. In to develop new management strategies against plant pathogens which are less toxic to non-targeted beneficial organisms and less persistent in nature, the objective of this study.is to assess the antifungal activity of various essential oils (EOs) and their major compounds against R. solani and F. solani which are responsible for causing damping off in common bean (Phaseolus vulgaris). The EOs used was extracted from cinnamon (Cinnamomum cassia), clove (Eugenia caryophyllata), lemongrass (Cymbopogon flexuosus), may chang (Litsea cubeba) and narrow-leaved peppermint (Eucalyptus radiata). In in vitro antifungal activity, potato dextrose agar (PDA) medium was mixed with these essential oils (EOs) at 50, 100, 200 and 400 µg/mL. Cinnamon EO displayed highest antifungal activity against tested pathogens among all the EOs, with 100% growth inhibition at the concentration 100 and 400 µg/mL, respectively. This was followed by lemongrass oil for R. solani and clove oil for F. solani. Eucalyptus oil was found ineffective against both fungi. The chemical analysis of essential oil through Gas chromatography/mass spectrometry (GC/MS) and bioassay indicated trans-cinnamaldehyde (74.04%) in cinnamon, eugenol (84.56%) in clove and geranial (36.72%) and neral (28.81%) in lemongrass as the major compounds which might be responsible for their highest inhibitory activities. The major compound of cinnamon oil; trans-cinnmaldehyde was most potent in inhibiting the mycelium of R. solani and F. solani at 50 µg/mL in vitro assay. In greenhouse, three of the most effective EOs and their respective major compounds were tested by seed treatment at 0.1% and 1% concentrations and by soil treatment at 2% concentration in the R. solani infested soil. The sterile substrate was infected with the inoculum of the R. solani at 2% (w/w). Cinnamon at 0.1% and 1% concentration as seed treatment demonstrated a good effect against the invasion of R. solani. The post-emergence disease incidence with this oil ranged from 24% to 46% and the severity percentage was between 24% to 38% and gave the survival of 40% to 60% at 0.1% and 1% respectively. Soil treatment with cinnamon oil was even better than seed treatment for reducing the percentage of disease incidence and severity and also improving the agronomical characters in bean. These results suggested that cinnamon, lemongrass and clove oil along with their active chemical constituents possess a strong antifungal activity against R. solani and F. solani whereas for the in vivo conditions, only cinnamon oil applied as soil treatment had good effect. Therefore, cinnamon oil can be a used as an alternative source of antifungal agent which has a good potential for improving the.control.of.damping.diseases.in plants and could be developed as the substitute to chemical pesticides to lower the negative impacts on the human health and environment.
參考文獻
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