Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/89292
標題: Effect of leaf essential oil and methanolic extracts from Eucalyptus citriodora on the growth of Mikania micrantha and its inhibition mechanisms
檸檬桉葉精油及甲醇抽出物抑制小花蔓澤蘭生長之效果及機制
作者: Yu-Jen Chen
陳宥任
關鍵字: 小花蔓澤蘭;植物相剋作用;檸檬桉;抗氧化酵素;單萜類含氧化合物;精油;Mikania micrantha;allelopathy;Eucalyptus citriodora;antioxidant enzyme;oxygenated monoterpenes;essential oil
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摘要: 
相剋作用(Allelopathy)一詞廣泛地用來描述兩植物間之化學交互作用,另外也被用在微生物之間、微生物與植物間和植物與昆蟲或食草動物之間的化學交流,可應用此作用作為清除或管理雜草的方法。本研究探討檸檬桉(Eucalyptus citriodora)葉子精油與甲醇抽出物之主要活性成分對小花蔓澤蘭(Mikania micrantha)之相剋機制。
檸檬桉葉子精油經 GC/MS 分析後,其主要成分為單萜類含氧化合物,選擇 Citronellal 、 Isopulegol 、 Citronellol 、 1,8-Cineole 與對照化合物 Eugenol 和 (-)-Carveol 以適當濃度進行小花蔓澤蘭生長抑制試驗。檸檬桉甲醇抽出物正己烷可溶部經管柱層析後得一具高活性之次分離部 H6-64 ,經試驗後發現其對於小花蔓澤蘭生長具有顯著抑制活性,以高效能液相層析儀分離純化後,經核磁共振光譜儀鑑定為 cis-p-Menthane-3,8-diol 和 trans-p-Menthane-3,8-diol,選擇 cis-p-Menthane-3,8-diol以不同濃度進行抑制試驗,發現其對小花蔓澤蘭生長具有顯著抑制活性,之後測量處理樣本中所含之過氧化氫與抗氧化酵素活性。
結果顯示各化合物在適當濃度下對小花蔓澤蘭幼苗生長仍具顯著抑制活性。胚根的抑制,精油之化合物之 IC50 均小於 5 ppm,在濃度為 10 ppm 時,其抑制率為 80—90%。cis-p-Menthane-3,8-diol 為 28 ppm,在濃度 100 ppm 以上可達到 90% 以上的抑制率。為了探討其抑制機制,在選擇濃度下,各化合物對小花蔓澤蘭胚軸生長抑制活性有限,當濃度為10 ppm 時,Citronellol 的胚軸抑制率為 55.6%,其他精油化合物皆小於50%。cis-p-Menthane-3,8-diol濃度 100 ppm時,胚軸生長抑制率為 49%。


檢驗其抑制機制發現,各化合物均會使小花蔓澤蘭之過氧化氫含量增加,此值與化合物之相剋活性和濃度呈正相關。在濃度 10 ppm 下,Citronellal 、 Citronellol 和 Isopulegol 分別為控制組的 219% 、 197% 和 149% 。而 cis-p-Menthane-3,8-diol 在濃度 300 ppm 時,為控制組的 141%。抗氧化酵素活性試驗結果也發現各處理組都會使小花蔓澤蘭胚根之抗壞血酸過氧化酵素(Ascorbate peroxidase, APX)與穀胱甘肽還原酵素(Glutathione reductase, GR)活性明顯提升。 cis-p-Menthane-3,8-diol在濃度為 10 ppm 時,小花蔓澤蘭胚根之 APX 活性與 GR 活性分別為控制組的 166% 和 207%。另外,各化合物均不會對過氧化氫酵素(Catalase, CAT)活性造成明顯提升,推測是被癒創木酚過氧化酵素(Guaiacol peroxidase, GPX)所取代,但由於本研究並無對 GPX 活性進行測定,無法確認 GPX 與 CAT 之相關,尚待後續研究追認。
綜合上述試驗結果,檸檬桉葉精油活性成分和 cis-p-Menthane-3,8-diol 對小花蔓澤蘭胚根生長具有高抑制活性,會誘導其胚根之過氧化氫含量大量增加,造成氧化壓力,破壞植物細胞與生長,達到生長抑制的功能。

Allelopathy is most commonly used to describe the chemical interaction between two plants, it has also been used to describe microbe-microbe, plant-microbe and plant-insect or plant-herbivore chemical communication. Allelopathy can be applied for weed removal. In this study we examined the allelopathic mechanism of growth suppression of Mikania micrantha by the major allelochemicals of Eucalyptus citriodora leaf essential oil and methanolic extracts.
Results of GC/MS analysis the major components of E. citriodora leaf essential oil was mainly as oxygenated monoterpenes. We choose citronellal, isopulegol, citronellol, 1,8-cineole, eugenol and (-)-carveol to conduct the growth suppression experiment at the concentrations of 1, 2 , 5 and 10 ppm. The subfraction H6-64 was isolated from n-hexane soluble fraction of E. citriodora leaf methanolic extracts, and it would inhibit the growth of radicle of M. micrantha effectively. Result of NMR analysis, the major component of H6-64 was cis-p-menthane-3,8-diol and trans-p-menthane-3,8-diol. We choose cis-p-menthane-3,8-diol to conduct the growth suppression experiment at the concentrations of 10, 20, 50, 100 and 300 ppm, and assess hydrogen peroxide concentration and antioxidant enzyme activity of M. micrantha seedling.
The results indicated that the major allelochemicals of E. citriodora leaf essential oil and cis-p-menthane-3,8-diol had high growth inhibition efficacy on M. micrantha seedling. The IC50 values of radicle growth inhibition were all < 5 ppm by allelochemicals in esstential oil, and cis-p-menthane-3,8-diol was 28 ppm. At a dosage of 10 ppm, the suppression rates of radicle growth were between 80% and 90% by allelochemicals in esstential oil, and the other were over 90% by cis-p-menthane-3,8-diol at the concentration of 100 ppm. However, the plumule growth inhibition by these allelochemicals were not so remarkable. When citronellol at the dosage of 10 ppm, the suppression rate was 55.6%, whereas others < 50%.
H2O2 concentration of treated M. micrantha would increased by these allelochemicals. H2O2 concentration was related to inhibitory bioactivitiy and dose. At a dosage of 10 ppm, citronellal, citronellol and isopulegol led to 219%, 197% and 149% increase in H2O2 concentration, respectively. On the other hand, cis-p-menthane-3,8-diol led to 141% increase at the concertration of 300 ppm. Results of antioxidant enzyme activity assay suggest showed that the allelochemicals increased ascorbate peroxidase activity and glutathione reductase activity of M. micrantha radicle. However, there was no significant increase of catalse activity.
The overall experimental results indicated that on the inhibitory effects of cis-p-menthane-3,8-diol and oxygenated monoterpenes of essential oil. These allelochemicals had strong growth inhibition efficacy against M. micrantha. These allelochemicals would increase the concentration of H2O2 of M. micrantha, and caused oxidative stress.
URI: http://hdl.handle.net/11455/89292
其他識別: U0005-2401201417282000
Rights: 同意授權瀏覽/列印電子全文服務,2016-01-28起公開。
Appears in Collections:森林學系

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