KIB OpenIR  > 昆明植物所硕博研究生毕业学位论文
Alternative TitleStudies on the Chemical Constituents of Cultured Fungal Endophytes from Medicinal Plants and Co-cultured Endophytic Fungi with Plant Cells
Thesis Advisor沈月毛
Degree Grantor中国科学院昆明植物研究所
Place of Conferral昆明植物研究所
Degree Discipline植物学
Keyword药用植物 内生真菌 化学成分 共培养 愈伤组织
Abstract本论文以从药用植物内生真菌中寻找活性化合物为主线,以云南美登木和喜树中分离得到的内生真菌为研究对象,从内生真菌纯培养及其与植物细胞共培养的化学成分研究两方面开展工作,取得了以下结果: 在对四株来源于喜树及云南美登木内生真菌的化学成分研究中,我们分离得到四十余个化合物,结构类型涉及单萜、倍半萜、降二萜、二倍半萜、聚酮、生物碱及麦角甾醇类,其中新化合物18个;并从中筛选到有抗肿瘤,抗真菌和抗细菌活性的化合物。具体为:从喜树的内生真菌XZ-07菌株的液体发酵产物中分离得到14个化合物,新化合物9个,其中化合物1为单萜,化合物2-9均为聚酮类。部分化合物进行了抗Hela细胞活性及抗菌活性实验,化合物11显示出中等的抗肿瘤活性及抗痢疾杆菌活性(Shigella dysenteriae);从云南美登木内生真菌MHF菌株的两种培养基的发酵产物中分离得到了20个化合物,botryosphaerin A-E (1-5)为新化合物,其中化合物1-4为四降劳丹烷型二萜,化合物5为劳丹烷型二萜。部分化合物进行了抗菌活性筛选,化合物12对白色念珠菌(Candida albicans)和酿酒酵母(Saccharomyces cerevisiae)的抑制作用与阳性对照制霉菌素相当;从云南美登木内生真菌Lz42菌株的固体发酵物中分离得到两个倍半萜和五个麦角甾醇类化合物,其中4-deacetyl-10-oxo-dihydrobotrydial (1)为一个新的倍半萜,化合物2为新的麦角甾醇类;从云南美登木内生真菌Ly22菌株的琼脂平板发酵物中分离得到4个化合物,其中3,4-epoxy-terpestacin (1)为一个新的二倍半萜,isoshihunine A (2)为一个新生物碱。 我们还建立了云南美登木愈伤组织与云南美登木内生真菌的共培养体系,筛选了七株云南美登木内生真菌,从中选择了Alternaria alternata Ly83与愈伤组织进行共培养。在TLC检测和抗菌活性测试的基础上,从纯培养和共培养的真菌Ly83中分离鉴定了26个化合物,结构类型主要包括混合萜、聚酮、环肽和麦角甾醇类,其中新化合物10个;并在实验结果的基础上进行了共培养的补充实验。此外,我们还从纯培养的愈伤组织中分离鉴定了16个已知化合物。结构类型主要包括齐墩果烷型、乌苏烷型及木栓烷型三萜和谷甾醇类。 最后,在综述部分总结了截止到2008年报道的从植物内生真菌中分离得到的有生物活性的次生代谢产物。
Other AbstractThe initiative of this dissertation was to search for new bioactive secondary metabolites from endophytic fungi of medicinal plant. On the basis of bioassay results, endophytes cultivated on Camptotheca acuminate and Maytenus hookeri were used as experimental material, and research works were carried out on the isolation, identification and activity detection of chemical components from cultured fungal endophytes and co-cultured endophytic fungi with plant cells. The main findings are as follows. More than forty compounds, including eighteen new ones, were obtained from the cultures of four endophytic fungi on C. acuminate and M. hookeri. Their structures involved terpenoids, polyketides, alkaloids and ergosterols, and some of them were bioactive. Concretely, 14 compounds comprising a new monoterpernoid and eight new polyketides were isolated from the liquid cultured endophyte Diaporthe sp. XZ-07 of C. acuminate, and compound 11 showed moderate cytotoxicity against human cancer Hela cells and antibacterial activity against Shigella dysenteriae. The investigation on secondary metabolites of endophytic fungal strain Botryosphaeria species MHF of M. hookeri, incubated on potato-dextrose-agar and Murashige-Skoog media, resulted in the isolation of five new terpenoids, botryosphaerins A-E (1-5), including four tetranorlabdanes and a labdane-type diterpene, together with fifteen known compounds. Among them, only compound 12 showed the similar inhibitory zones against Candida albicans and Saccharomyces cerevisiae as the positive control, nystatin. From solid cultures of endophyte Phomopsis species Lz42 of M. hookeri, a new sesquiterpenoid, 4-deacetyl-10-oxo-dihydrobotrydial (1), and a new sterol, 22E,24R-5α,8α- epidioxyergosta-6,22-dien-3β-formate (2), were isolated together with five related known compounds. We also studied the chemical constituents of endophytic fungal Colletotrichum sp. Ly22 from M. hookeri and gained 4 compounds, including a new sesterterpenoid and a new alkaloid. To study the interactions between endophytes and their host plants, the co-culture of Alternaria alternata Ly83, one of endophytic fungi cultivated on M. hookeri, and callus of M. hookeri was established on the basic principle of chemical defense. The chemical study of the solid cultures and co-cultures of A. alternata led to the isolations of 26 compounds, including 9 new mixed terpenoids and a new polydetide. Based on the results, complementary experiments for co-culture were performed. In addition, 16 known compounds, mainly consisted of terpenoids and sitosterols, were isolated from the callus of M. hookeri. In the last part of this dissertation, a review covers 106 literatures until 2008 and 173 bioactive compounds isolated from plant endophytic fungi were presented.
Document Type学位论文
Recommended Citation
GB/T 7714
袁琳. 植物内生真菌及与植物细胞共培养的化学成分[D]. 昆明植物研究所. 中国科学院昆明植物研究所,2008.
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