Institute of Chinese Herbal Medicines, Hubei Academy of Agricultural Sciences, No. 253 Xueyuan Road, Enshi 445000, China.
Institute of Chinese Herbal Medicines, Hubei Academy of Agricultural Sciences, No. 253 Xueyuan Road, Enshi 445000, China.
J Infect Public Health. 2021 Dec;14(12):1917-1926. doi: 10.1016/j.jiph.2021.10.019. Epub 2021 Oct 25.
Serious infections caused by bacteria and their resistance to antibiotics are one of the biggest healthcare threats to mankind. Therefore, the present study aimed to isolate endophytes from medicinal plant Ampelopsis grossedentata, an endemic species of Western Hubei, China and to investigate its antibacterial efficacy and chemical diversity of the secondary metabolites.
The antibacterial potential of the endophytes was evaluated by disc diffusion method against a panel of eleven type strains and some multidrug resistant pathogenic bacteria. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were estimated by broth microdilution using iodonitrotetrazolium chloride assay. Further, the chemical diversity of the metabolites was estimated using LC-Q-TOF-MS/MS and GC-MS fingerprinting.
Four endophytic fungi were isolated from the tender shoot of A. grossedentata; they were identified as Fusarium graminearum TC-1, Phomopsis mali TC-3, Pestalotiopsis maculans TC-5 and Alternaria alternata TC-11. Among the endophytes screened, A. alternata TC-11 exhibited significant antibacterial activity with the zones of inhibition ranging from 13.72 ± 0.30 to 21.76 ± 0.53 mm against all the tested type strains and multidrug resistant bacterial pathogens. Further, it showed significant antibacterial activity with MIC values ranging from 0.37 to 3.00 μg/mL. The combined LC-Q-TOF-MS/MS and GC-MS analyses of active extract revealed that alternarian acid, altertenuol, dimethyl sulfone, docosane, dodecane, duclauxin, ergosta-4,6,8(14),22-tetraen-3-one, ethyl 6-cyano-5-oxo-1-phenyl-7-thiophen-2-yl-[1,2,4]triazolo[4,3-a]pyrimidine-3-carboxylate, heptacosane, linoleic acid, neodecanoic acid, oxiranylmethyl ester, pentadecane, verrulactone E, 2,6,11-Trimethyldodecane and 4-[(E,4R,6R)-11-(furan-3-yl)-6-hydroxy-4,8-dimethylundec-8-enyl]-2-hydroxy-2H-furan-5-one were the most abundant compounds present which were responsible for the significant antibacterial activity.
To our knowledge, this is the first report of fungal endophytes isolated from the tender shoot of A. grossedentata with bacteriostatic and bactericidal activities. Our finding provides a new insight into the antibacterial potential of endophytes and envisages the possibility of using them for drug discovery.
由细菌引起的严重感染及其对抗生素的耐药性是人类面临的最大健康威胁之一。因此,本研究旨在从中国湖北西部特有种药用植物 Ampelopsis grossedentata 的内生菌中分离内生菌,并研究其抗菌功效和次生代谢物的化学多样性。
采用纸片扩散法对 11 株标准菌株和部分多药耐药病原菌进行抗菌活性评价。采用碘硝基四唑氯化物法通过肉汤微量稀释法估计最小抑菌浓度(MIC)和最小杀菌浓度(MBC)。此外,采用 LC-Q-TOF-MS/MS 和 GC-MS 指纹图谱估计代谢物的化学多样性。
从 Ampelopsis grossedentata 的嫩枝中分离出 4 株内生真菌,它们分别被鉴定为禾谷镰刀菌 TC-1、苹果茎点霉 TC-3、交链孢霉 TC-5 和Alternaria alternata TC-11。在所筛选的内生菌中,Alternaria alternata TC-11 对所有测试的标准菌株和多药耐药病原菌均表现出显著的抗菌活性,抑菌圈直径为 13.72±0.30-21.76±0.53mm。此外,它对 MIC 值的抗菌活性范围为 0.37-3.00μg/ml。活性提取物的 LC-Q-TOF-MS/MS 和 GC-MS 联合分析表明,Alternaria 酸、altenuol、二甲亚砜、二十二烷、十二烷、duclauxin、麦角甾-4,6,8(14),22-四烯-3-酮、乙基 6-氰基-5-氧代-1-苯基-7-噻吩-2-基-[1,2,4]三唑[4,3-a]嘧啶-3-羧酸酯、十七烷、亚油酸、正癸酸、环氧甲酰基甲酯、十五烷、verrulactone E、2,6,11-三甲基十二烷和 4-[(E,4R,6R)-11-(呋喃-3-基)-6-羟基-4,8-二甲基十一-8-烯基]-2-羟基-2H-呋喃-5-酮是含量最丰富的化合物,这些化合物负责显著的抗菌活性。
据我们所知,这是首次从 Ampelopsis grossedentata 的嫩枝中分离出具有抑菌和杀菌活性的真菌内生菌。我们的发现为内生菌的抗菌潜力提供了新的见解,并预示着它们在药物发现中的应用可能性。