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目的:药用植物内生真菌是一类重要的微生物资源,其代谢产物有着广泛的生物学活性。该研究拟从野葛(Pueraria lobata(Willd.)Ohwi)中分离有细胞毒活性的内生真菌。方法:组织块改良法分离内生真菌;细胞毒活性测定采用Alamar blue法;结合形态学和分子生物学方法进行菌种鉴定。结果:获得的34株内生真菌中菌株KLBMP f0027对HepG2、HO-8910、NCI-H460、SGC-7901等细胞株均有显著活性,ID50分别为437.4、460.0、542.5、771.2。而且活性产物相对稳定,对温度、pH变化及蛋白酶不敏感。代谢过程研究显示菌株KLBMP f0027的活性产物合成与细胞生长属于部分耦联关系类型。形态学和ITS-rDNA序列分析鉴定菌株为Aspergillus ostianus strain KLBMP f0027。结论:野葛内生真菌A.ostianus strain KLBMP f0027可作为细胞毒活性候选菌株进行深入研究。
Objective: Endophytic fungi of medicinal plants are a kind of important microbial resources, and their metabolites have a wide range of biological activities. The study was designed to isolate cytotoxic endophytic fungi from Pueraria lobata (Willd.) Ohwi. Methods: Tissue block method was used to separate endophytic fungi. Alamar blue method was used to determine the cytotoxicity. The morphological and molecular biological methods were used to identify the bacteria. Results: Among the 34 strains of endophytic fungi, KLBMP f0027 had significant activity on HepG2, HO-8910, NCI-H460 and SGC-7901 cell lines with ID50 of 437.4, 460.0, 542.5 and 771.2, respectively. And the active product is relatively stable, temperature, pH changes and protease insensitivity. Metabolic process studies show that the synthesis of the active product of strain KLBMP f0027 is part of the coupling relationship with cell growth. Morphological and ITS-rDNA sequence analysis identified strain Aspergillus ostianus strain KLBMP f0027. CONCLUSION: A. utilis strain KLBMP f0027, an endophytic fungus of kudzu can be further studied as a cytotoxic candidate strain.