內生真菌已被證實存在於所有已知植物種類中，其對植物宿主明顯地有著重要的角色。然而，這類真菌的多樣性和抗微生物活性仍大部分不清楚。在本研究中，吾人由距離中國東南海岸線約2公里的小金門島，採集5個藥用紅樹林植物老鼠?(Acanthus ilicifolius var. xiamenensis)樣本，從其中篩選出具有生物活性的內生真菌。吾人使用三種不同的選擇性培養基分離培養樣品中所含內生真菌，總共從葉子樣品中分離出579株真菌。通過聚合?連鎖鏈反應(polymerase chain reaction)擴增這些真菌菌株的rDNA內轉錄間隔區(internal transcribe sequences, ITS)片段並定序，成功定序出208條序列。其BLAST結果顯示它們分屬於三個門、二十六個科和三十三個屬。這些生長在紅樹林植物中的內生真菌菌株主要為Fusarium屬(40株, 19.4%)、Didymella屬(19, 9.2%)和Allophoma屬(17, 8.3%)。分離的內真菌使用洋菜錠法對六種致病菌進行抑制活性測試。在所有測試菌株(243株)中，41.15% (100株)菌株顯示出對一種或多種致病菌有抑制活性的效果。接著再進行第二次三重複抑菌測試，結果顯示為62株具有抑制活性，其中Fusarium屬(12株)、Epicoccum屬(8株)和Allophoma屬(6株)是具有抗菌活性真菌菌株的三個主要屬，佔有抑制效果菌株的42%。6個分離株，即LS-04-05-sM-3、LS-04-01-sM-6-1、LS-05-01-sP-4、LS-03-04-sG-6、LS-04-02-eM-3和LS-04-02-eM-5，表現出強或對多種致病菌有抑菌活性，這幾株可做為未來進一步天然物分離和研究。在本研究顯示，紅樹林植物老鼠?的內生真菌有高的多樣性，紅樹林植物內生真菌可以是天然抑菌劑的來源；這些能產生具有抑制活性的植物內生真菌可能在組織中與宿主之間相互作用並有重要的生態作用。

Fungal endophytes have been found to exist in every plant species examined to date and appear to be important to their plant hosts. However, the diversity and antimicrobial activity of this group of fungus are largely unknown. In present research, five leave samples, collected from mangrove plant Acanthus ilicifolius var. xiamenensis at Lesser Kinmen Island, located roughly 2 km from the southeastern coastline of China, were used to screen for bioactive metabolites producing endophytic fungi. Endophytic fungal strains were isolated by using agar-based culture technique with three different selective media. A total 579 fungi were isolated from the leaf samples. The rDNA internal transcribed spacer (ITS) fragments of 208 (35.92%; 208 out of 579) fungal strains were amplified by polymerase chain reaction and sequenced. BLAST results suggested that they belong to three phyla, twenty-six families, and thity-three genera. The fungal strains inhabiting the mangrove plant were predominantly identified as Fusarium (40 strains, 19.4%), Didymella (19, 9.2%), and Allophoma (17, 8.3%). The antimicrobial activity of isolated fungi was performed against six indicator microorganisms by agar block methods. Among all the tested strains (243), 41.15% (100) of the strains showed antimicrobial activity against one or more of the indicator microorganisms. Fusarium (12 strains), Epicoccum (8), and Alternaria (6) were the three major genera with antimicrobial activity, accounting for 42% of the antibiotic-producing strains. six isolates, namely LS-04-05-sM-3, LS-04-01-sM-6-1, LS-05-01-sP-4, LS-03-04-sG-6, LS-04-02-eM-3 and LS-04-02-eM-5which exhibited strong or wide-spectrum antimicrobial activity were good candidates for further natural product isolation and characterization research. Our study demonstrated that the endophytic fungal diversity in mangrove plant Acanthus ilicifolius var. xiamenensis was high. Endophytic fungi from mangrove plant can be a source of natural antimicrobial agents; and these antibiotic-producing fungi may also play an important role in the ecological interaction between endophyes and their host.

第一章 緒論 1
第二章 實驗材料與方法 13
第三章 結果 27
第四章 結論 75
參考文獻 79
附錄 89

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