【作者】 王丽薇；

【导师】 林福呈； 章初龙；

【作者基本信息】 浙江大学 ， 植物病理学， 2011， 博士

【摘要】 植物内生真菌是人类尚未充分开发利用的真菌资源之一,内生真菌产生的次生代谢产物种类繁多,在农业、医药等领域具有重要的应用潜力。本论文对六种药用植物内生真菌进行了分离,获得菌株112株,离体活性筛选获得22株具有抗菌活性的菌株。以其中6株抗菌活性较强、抗菌谱较广的菌株为研究对象,对其次生代谢产物进行活性跟踪分离,获得18个化合物,鉴定了其中15个化合物,其中1个为新化合物,1个为新天然产物,另有5个化合物为首次从菌属中分离获得的天然产物,1个化合物为首次从菌种中分离获得的天然产物。对其中部分化合物进行抗菌、抗肿瘤活性研究。从天南星内生真菌茎点霉Phoma sp. ZJWCF006的次生代谢产物中分离获得6个化合物,鉴定了5个化合物：(3S)-3,6,7-三羟基-α-四氢萘酮(1)、尾孢酰胺(2)、木霉菌素(3)、β-谷甾醇(4)、富马酸单乙酯(5)。其中1为新化合物,化合物3-5均为首次从茎点霉属内生真菌中分离获得。抗菌活性测试结果表明,化合物2对十种植物病原菌均有较强抑制作用。抗肿瘤测试结果表明化合物2对六种肿瘤细胞有中等抑制作用。从马兰内生真菌茎点霉Phoma sp. ZJLQ335中分离并鉴定了1个化合物：尾孢酰胺(2)。在此基础上,对该内生真菌发酵培养基进行优化实验,使其中主要产物尾孢酰胺(2)的产率提高了9.32倍。从多花黄精内生真菌变灰青霉Penicillium canescens zjqy610中分离并鉴定了3个化合物：邻乙酰苯基脒基甲酸(6)、灰黄霉素(7)和4-羟基-5-甲氧基-2-甲基萘[1,2-b]呋喃-3-甲酸(8)。其中化合物8为首次从微生物中分离获得的新天然产物,化合物6为首次从内生真菌青霉属中分离获得。抗菌活性测试表明,化合物7对十四种植物病原真菌都具有抑制活性。从银杏内生真菌团青霉Penicillium commune TMSF169中分离并鉴定了一个化合物圆弧菌醛酸(9),该化合物为首次从团青霉中分离获得。抗菌活性测试表明,化合物9对十二种植物病原真菌具有抑制活性。从无花果果内生真菌炭角菌Xylaria sp. ZJWCF255中分离并鉴定了4个化合物：细胞松弛素D(10)、细胞松弛素C(11)、细胞松弛素Q(12)与细胞松弛素R(13)。抗菌活性测试表明,化合物12对十五种植物病原真菌具有抑制活性。抗肿瘤测试结果表明化合物12对三种肿瘤细胞有中等抑制作用。从博落回内生真菌Chaetomium cupreum ZJWCF079中分离了4个化合物,鉴定了其中2个化合物：3,3’,6,6’-四羟基-4,4’-二甲基-1,1’-二(环己一3,6二烯)-2,2’,5,5’-四酮(14)、麦角甾烷-5,7,22-三烯-3-醇(15),其中化合物15为首次从毛壳菌属内生真菌中分离获得。本论文的结果表明,药用植物内生真菌次生代谢产物具有结构新颖性与生物活性多样性的特征,是寻找先导化合物重要资源。更多还原

【Abstract】 In this dissertation, six medicinal plants were picked as research sources.112different fungal strains were separated from these medicinal plants and screened on antifungal models,22strains showed antifungal activities were obtained. Among which, six strains were selected for further study because of their higher and broader spectrum of antifungal activities. Bioactive-guided isolation and elucidation of the metabolites of five strains obtained18compounds.15compounds were identified and among which one was a new compound, one was a new natural product, the other five compounds were isolated from the endophytic genus for the first time and still another one was obtained from the endophytic species for the first time.From the extracts of Phoma species ZJCWF006in Arisaema erubescens (Wall) Schott., six compounds were separated and two of them were elucidated as (3S)-3,6,7-trihydroxy-α-tetralone (1), cercosporamide (2), trichodermin (3), β-sitosterol (4). Monoethyl fumarate (5). Among which compound1was a new a-tetralone derivative, compound3-5were isolated from Phoma sp. for the first time. Bioactivity assays showed that compound2showed strong antifungal activities against ten tested plant pathogenic fungi. Compound2also exhibited moderate cytotoxicity against the six tumor cell lines tested. From the extracts of Phoma sp. ZJLQF335in Kalimeris indica (Linn.) Sch.-Bip., one compound was isolated and elucidated as cercosporamide (2). Optimization experiment of fermentation medium of the fungal strain resulted in yield increasing of the major metabolite2to9.32times than before.From Penicillium canescens zjqy610in Polygonatum cyrtonema Hua, three compounds were isolated and structure elucidated, including o-Acetylbenzeneamidinocarboxylic Acid (6), Griseofulvin (7) and Naphtho[1,2-b]furan-3-carboxylic acid,4-hydroxy-5-methoxy-2-methyl-(8). Among which, compound8was a new natural product from microbiology, compound6was obtained from Polygonatum sp. for the first time. Bioactivity assays showed compound7showed antifungal activities against fourteen tested plant pathogenic fungi.One compound with antifungal activities was separated from Penicillium commune TMSF169residing in Ginkgo biloba Linn. and identified as Cyclopaldic acid (9). It was isolated from P. commune for the first time. Bioactivity assays showed that compound9showed antifungal activities against twelve tested plant pathogenic fungi.From the extracts of Xylaria sp. ZJWCF255in Lycopersivon esculentum Mill. Four compounds were isolated and elucidated as cytochalasin D (10), cytochalasin C (11), cytochalasin Q (12), cytochalasin R (13). Bioactivity assays showed that compound12showed antifungal activities against fifteen tested plant pathogenic fungi.The secondary metabolites of the strain Chaetomium cupreum ZJWCF079in Macleaya cordata (Willd.) R.B was studied. Four compounds were isolated and two of them were elucidated as3,3’,6,6’-tetrahydroxy-4,4’-dimethyl-1,1’-bi(cyclohexa-3,6-diene)-2,2’,5,5’-tetraone (14) and Ergosterol (15) respectively. Compound15were isolated from Chaetomium sp. for the first time.Our study therefore underscores that endophytic fungi in medicinal plants are promising sources of natural bioactive and novel metabolites.更多还原