滇重楼内生真菌的分离及其抑菌活性
Isolation and Antibacterial Activity of Fungal Endophytes in Paris polyphylla Smith var. yunnanenesis
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摘要: 目的 从滇重楼新鲜植株中分离内生真菌并研究其代谢产物抑制人体病原细菌的能力.方法 滇重楼新鲜植株表面消毒后, 采用YPD平板分离培养内生真菌.各菌株的液体发酵物经乙酸乙酯萃取并浓缩.然后, 以金黄色葡萄球菌、伤寒沙门菌、屎肠球菌3种可感染人体的细菌为指示菌, 采用滤纸片琼脂扩散法测定抑菌活性.结果 分离到54株内生真菌.其中20株内生真菌有抑菌活性, 活性菌株占总菌株数量的37%.其中, J09对上述3种细菌的抑菌圈直径分别为 (49.67±1.25) mm、 (50.33±0.52) mm、 (37.00±1.63) mm, 经形态学观察及分子生物学鉴定, J09为灰黄青霉.同时, 经高效液相色谱检测确定, 该抑菌成分并非青霉素或灰黄霉素.结论 内生真菌 (J09) 的代谢产物抑菌活性较强, 具有开发抗菌药物的潜在价值.Abstract: Objective To isolate the fungal endophytes from Paris Polyphylla Smith var.yunnanensis and investigate the antibacterial activities of these fungi to human's pathogenic bacteria. Methods After surface sterilization, fungal endophytes were cultivated on yeast powder dextrosed (YPD) .The fermentation liquid of every fungal strain was extracted by ethyl acetate. Afterwards, the effect of antibacterial activity on Staphylococcus aureus, Salmonella typhi and Endococcus faecium was investigated by paper disk test. Results54 strains of fungal endophytes were isolated and purified. Among them, 20 strains had antibacterial activities, which accounted for 37% of the total isolated strains. Inhibition zones of J09 metabolites to the three indicators was ( 49.67 ±1.25) mm, ( 50.33 ±0.52) mm, ( 37.00 ±1.63) mm respectively. In terms of the morphological and molecular biological characteristics, J09 was identified as Penicillium griseofulvum.Meanwhile, the antibacterial compounds were not penicillin or griseofulvin by high performance liquid chromatography (HPLC) detection. Conclusion The fungal endophyte J09 could be a potential candidate to develop an antibacterial medicine since its metabolites have showed strong antibacterial activities.
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