細辛抑制木黴菌生長活性成份研究

現今可商業化栽培之食用菇類有20幾種，木黴菌為菇類培養初期主要的污染微生物，導致農藥使用與成本增加，降低菇類的產值，因此防治木黴菌污染應可提高菇類的產量與品質。細辛(Asarum heterotropides Fr. Schmidt var. mandshuricum (Maxim.) Kitag.) 為馬兜鈴科細辛屬細辛的根部，具有祛風散寒、止痛、溫肺化飲、止咳、開竅的功效。本研究以中藥草萃取物進行木黴菌(Trichoderma harzianum)之菌絲生長抑制活性篩選時，發現細辛乙醇粗萃物在2000 ppm能抑制 T-2-1、T-3-1及T-A-1菌絲生長分別為 56.8±0.06、58.7±0.06及53.1±0.10％，因此將細辛乙醇粗抽物進行極性分配萃取並活性篩選，其中CHCl3層在2000 ppm時對T-2-1、T-3-1及T-A-1可達到64.3±0.00、65.5±0.00和56.0±0.00％的菌絲生長抑制作用。依T. harzianum菌絲生長抑制活性為導向，利用管柱層析及再結晶等方法進行細辛成分分離，得到九個化合物。經文獻比對各化合物的光譜數據，確認7個化合物構造為β-sitosterol (1), (-)-asarinin (2), (-)-sesamin (3), (-)-pluviatilol (4), methyl 3,4-dihydroxybenzoate (5), aristololactam I (6), methylkauol (7)。將分離得到之化合物Ashe-1, 2和3進行T. harzianum T-2-1、T-3-1及T-A-1菌絲生長與孢子萌發抑制活性試驗，顯示三個化合物在100 ppm時對T-2-1、T-3-1及T-A-1均具有孢子萌發抑制活性達61-79％，500 ppm時均具有孢子萌發抑制活性達74-85％。且Ashe-1在40 ppm時具26.9%、Ashe-2在6 ppm時具13.4%及Ashe-3在40 ppm時具32.1%的菌絲生長抑制活性，由此結果得知，細辛萃取物具有抑制木黴菌生長活性，未來可進行田間試驗評估其開發為生物農藥的價值。

關鍵字

細辛；木黴菌；菇類；菌絲生長抑制；孢子萌發

並列摘要

More than 20 kinds of mushroom have been cultured for food and medicine. However, Trichoderma harzianum is one of major contamination fungi in the initial stage of mushroom cultivation. It leaded to use pesticide and costs increased. Therefore, to prevent contamination of T. harzianum should be able to improve the yield and quality of mushrooms. The dried roots of Asarum heterotropides Fr. Schmidt var. mandshuricum (Maxim.) Kitag., belong to Aristolochia family, are used for analgesic and anti-tussive in traditional Chinese medicine. In this study, we have examined several Chinese herbal extracts to inhibit the growth of T. harzianum (T-2-1, T-3-1 and T-A-1), and found that the ethanol extract of A. heterotropides var. mandshuricum exhibited 56.8±0.06, 58.7±0.06 and 53.1±0.10% growth inhibition activity at 2000 ppm. Then, the ethanol extract was partitioned into chloroform, n-butanol, and water layers, respectively. The chloroform layer exhibited 64.3±0.00, 65.5±0.00 and 56.0±0.00％ mycelial growth inhibition of T-2-1, T-3-1 and T-A-1 at 2000 ppm. According bioassay guiding result, the CHCl3 layer was isolated by column chromatography and recrystallization to obtain nine compounds. Their structures were elucidated by spectroscopic analysis and compared with the literature data, and identified as β-sitosterol (1), (-)-asarinin (2), (-)-sesamin (3), (-)-pluviatilol (4), methyl 3,4-dihydroxybenzoate (5), aristololactam I (6) and methylkauol (7). Compounds Ashe-1~3 were evaluated the inhibition activity of spore germination and mycelial growth against T. harzianum T-2-1, T-3-1 and T-A-1. The result showed that Ashe-1~3 exhitbited 61-79 and 74-85％ spores germination inhibition at 100 and 500 ppm, respectively. The mycelial growth inhibition activities of Ashe-1~3 were 26.9% of Ashe-1 at 40 ppm, 13.4% of Ashe-2 at 6 ppm, and 32.1% of Ashe-3 at 40 ppm, respectively. Based on the above results, A. heterotropides var. mandshuricum may be developed as biological pesticide against T. harzianum.