- 期刊
微量滴定板篩選植物萃取液與植物(精)油抑制菜豆銹病菌及草莓灰黴病菌
Microtiter Plate for Evaluation of Plant Extracts and Oils Against Uromyces appendiculatus and Botrytis cinerea
摘要
為尋找自然界潛在的植物源植物保護資材，本研究以微量滴定板（microtiter plate）搭配水瓊脂平板建立一套快速、精確的方法以供篩選對植物病原真菌具抑制效果的植物萃取液或植物(精)油。將供試之植物源資材與待測之草莓灰黴病菌（Botrytis cinerea）分生孢子或菜豆銹病菌（Uromyces appendiculatus）夏孢子均勻混入微量滴定板之盤穴中，覆以封口膜，靜置於24℃定溫箱中2小時。再將盤穴中的混合物分別塗佈於2%水瓊脂平板上，復於24℃定溫箱培養12小時，計數孢子發芽率以測定植物源植保資材的抑菌效果。試驗結果顯示，供試之85種植物萃取液之原液對草莓灰黴病菌均不具抑菌作用；但烏心石、台灣假黃楊、台灣光臘樹、菲律賓厚殼桂及翼柄決明等5種植物萃取液於稀釋10倍濃度下，對菜豆銹病菌尚具抑菌效果。供試之23種植物(精)油原液則有部份對上述兩種病原菌表現良好的抑菌能力。將這些初試有效的植物(精)油原液分別與無患子果實發酵液以等體積比例混合調製再加水稀釋，依上述方法測定其對這兩種植物病原菌之抑菌效果。結果顯示，稀釋1000倍之肉桂油及百里香精油製劑可完全抑制草莓灰黴病菌孢子發芽；單獨無患子果實發酵液於稀釋10倍即對該菌不具抑制作用。相同之試驗處理，稀釋1000倍之肉桂油、丁香油及百里香精油製劑可完全抑制菜豆銹病菌孢子發芽；單獨無患子果實發酵液於稀釋100倍即對該菌孢子發芽不具抑制作用。
並列摘要
A rapid and precise assay to determine antifungal activity in plant extracts and plant (essential) oils by using microtiter plate is described. This method would provide a very useful tool to screen numerous naturally antifungal materials against plant pathogens. Wells in microtiter plate were loaded with a mixture of spore suspension of Uromyces appendiculatus or Botrytis cinerea and plant extract or plant (essential) oil. The microtiter plate was then sealed with parafilm membrane and placed in 24℃ incubator for 2 hr. After that, the mixture in each well was spread on 2% water agar and incubated for another 12 hr at 24℃. The spore germination of the fungus was measured under microscope. Among 85 undiluted plant extracts evaluated, none of them possessed antifungal activity on B. cinerea, while 10-fold dilutions of plant extracts of Michelia compressa, Liodendron formosanum, Fraxinus formosana, Cryptocarya elliptifolia and Senna alata indicated antifungal activity well against U. appendiculatus. Among 23 plant (essential) oils tested, thyme essential oil and cinnamon oil demonstrated completely antifungal activity against both fungal pathogens when applying 1000-fold dilution of the formulation of the each oil and Chinese soap berry extract in 1:1 volume ratio. However, the same dilution of the formulated clove oil could only inhibit the spore germination of U. appendiculatus but not B. cinerea.