毛筒壳科真菌次级代谢产物生物活性的评价

毛筒壳科Tubeufiaceae真菌具有产新结构、新活性次级代谢产物的潜力,目前对该科真菌次级代谢产物的研究较少。为了寻找具有生物活性的新化合物,有必要对毛筒壳科真菌次级代谢产物及其活性进行系统深入的研究。本文采用平板对峙法、生长速率法和MTT法,分别测定已分离得到的19株该科真菌活体菌株抑菌活性、发酵物抑菌活性以及发酵物粗提物对不同人体肿瘤细胞株增殖的抑制作用。通过平板对峙法,试验共筛选获得13株活性菌株,其中,红棕毛筒腔菌菌株Tubeufia rubra PF02-2对7种植物病原真菌有明显的抑菌效果,抑制率均高于60%且抑菌谱广。采用生长速率法,发现红棕毛筒腔菌菌株PF02-2经液体发酵后,发酵液对其中4种植物病原真菌仍有一定的抑制作用,且菌丝体部分的乙酸乙酯提取物对马铃薯早疫病病菌Alternaria solani（ZYB）的抑制效果最好。通过MTT法,发现发酵物粗提物对3种肿瘤细胞均具有不同程度的细胞毒活性,其中在300μg/mL时,剑叶莎毛筒腔菌菌株Tubeufia machaerinae ML03-2发酵液部分的乙酸乙酯提取物对人宫颈癌细胞株HeLa和人前列腺癌细胞株PC-3的抑制率（%）分别达到了98.92±0.15和97.86±0.18,在400μg/mL时,对人肝癌细胞株HEPG2的抑制率（%）达到了98.88±0.04;在500μg/mL时,明孢新旋卷孢菌菌株Neohelicosporium hyalosporum ML05-1菌丝体部分的乙酸乙酯提取物对人宫颈癌细胞株HeLa细胞的抑制率（%）为98.32±0.02,在600μg/mL时,对人肝癌细胞株HEPG2的抑制率（%）达到了97.62±0.20,在300μg/mL时,对人前列腺癌细胞株PC-3的抑制率（%）达到了98.91±0.02。该研究结果为开发利用毛筒壳科真菌提供了科学依据。

Bioactivity evaluation of secondary metabolites produced by species of Tubeufiaceae

Secondary metabolites of fungi belonging to Tubeufiaceae are still imperfectly known. Through plate confrontation culture, colony growth rate determination and MTT method, 19 tubeufiaceous living strains and their fermentation substances were tested for observation of antifungal activities. The inhibition rate (IBR) of the crude fermentation extract against different cell lines of human tumors was also determined. Thirteen strains with obvious bioactivity were obtained. The strain Tubeufia rubra PF02-2 showed notable antifungal effects against seven plant pathogenic fungi with IBR over 60% and widest antifungal spectrum. The fermentation broth of Tubeufia rubra PF02-2 had certain inhibition effect on four plant pathogenic fungi. The ethyl acetate extract of mycelia exhibited excellent inhibition effect on potato early blight pathogen Alternaria solani. MTT assay indicated that crude extract of the fermentation broth of different fungal strains showed various degrees of cytotoxic activity against three human tumor cells. At 300μg/mL, the IBR (%) of ethyl acetate extract of Tubeufia machaerinae ML03-2 fermentation broth against human cervical cancer cell line HeLa cells and human prostate cancer cell line PC-3 cells reached 98.92±0.15 and 97.86±0.18, respectively, and at 400μg/mL, the IBR (%) of the extract against human hepatoma cell line HEPG2 cells reached 98.88±0.04. At 500μg/mL, the IBR (%) of Neohelicosporium hyalosporum ML05-1 mycelium ethyl acetate extract against human cervical cancer cell line HeLa cells was 98.32±0.02; at 600μg/mL, the IBR (%) of the extract against human liver cancer cell line HEPG2 cells reached 97.62±0.20, and at 300μg/mL, the IBR (%) against human prostate cancer cell line PC-3 cells was 98.91±0.02. The results of this study provide reference data for potential application value of tubeufiaceous fungi and the development possibility through pharmaceutical agrochemical industry.