Wei Longfeng, Ran Jiang, Li Zhu, Zhang Qinyu, Guo Kun, Mu Shuzhen, Xie Yudan, Xie Ailin, Xiao Yang
Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), College of Life Sciences, Institute of Agro-bioengineering, Guizhou University, Guiyang, Guizhou Province, 550025, China.
Guizhou Key Laboratory of Agricultural Biotechnology, Guiyang, 550009, China.
Appl Biochem Biotechnol. 2024 Feb;196(2):878-895. doi: 10.1007/s12010-023-04577-9. Epub 2023 May 31.
Six compounds were isolated and purified from the crude acetone extract of Aspergillus niger xj. Characterization of all compounds was done by NMR and MS. On the basis of chemical and spectral analysis structure, six compounds were elucidated as metazachlor (1), nonacosane (2), palmitic acid (3), 5,5'-oxybis(5-methylene-2-furaldehyde) (4), dimethyl 5-nitroisophthalate (5) and cholesta-3,5-dien-7-one (6), respectively, and compounds 1, 4, 5 and 6 were isolated for the first time from A. niger. To evaluate the antibacterial activity of compounds 1-6 against three plant pathogenic bacteria (Agrobacterium tumefaciens T-37, Erwinia carotovora EC-1 and Ralstonia solanacearum RS-2), and the minimum inhibitory concentrations (MICs) were determined by broth microdilution method in 96-well microtiter plates. Results of the evaluation of the antibacterial activity showed that T-37 strain was more susceptible to metazachlor with the lowest MIC of 31.25 µg/mL. The antibacterial activity of metazachlor has rarely been reported, thus the antibacterial mechanism of metazachlor against T-37 strain were investigated. The permeability of cell membrane demonstrated that cells membranes were broken by metazachlor, which caused leakage of ions in cells. SDS-PAGE of T-37 proteins indicated that metazachlor could damage bacterial cells through the destruction of cellular proteins. Scanning electron microscopy results showed obvious morphological and ultrastructural changes in the T-37 cells, further confirming the cell membrane damages caused by metazachlor. Overall, our findings demonstrated that the ability of metazachlor to suppress the growth of T-37 pathogenic bacteria makes it potential biocontrol agents.
从黑曲霉xj的丙酮粗提物中分离并纯化出六种化合物。所有化合物均通过核磁共振(NMR)和质谱(MS)进行表征。基于化学和光谱分析结构,六种化合物分别被鉴定为异丙甲草胺(1)、二十九烷(2)、棕榈酸(3)、5,5'-氧双(5-亚甲基-2-糠醛)(4)、5-硝基间苯二甲酸二甲酯(5)和胆甾-3,5-二烯-7-酮(6),其中化合物1、4、5和6首次从黑曲霉中分离得到。为了评估化合物1 - 6对三种植物病原菌(根癌土壤杆菌T - 37、胡萝卜软腐欧文氏菌EC - 1和青枯雷尔氏菌RS - 2)的抗菌活性,采用肉汤微量稀释法在96孔微量滴定板中测定了最低抑菌浓度(MIC)。抗菌活性评估结果表明,T - 37菌株对异丙甲草胺最为敏感,最低MIC为31.25 μg/mL。异丙甲草胺抗菌活性的报道较少,因此研究了其对T - 37菌株的抗菌机制。细胞膜通透性实验表明,异丙甲草胺破坏了细胞膜,导致细胞内离子泄漏。T - 37蛋白的十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)表明,异丙甲草胺可通过破坏细胞蛋白来损伤细菌细胞。扫描电子显微镜结果显示T - 37细胞出现明显的形态和超微结构变化,进一步证实了异丙甲草胺对细胞膜的损伤。总体而言,我们的研究结果表明,异丙甲草胺抑制T - 37病原菌生长的能力使其具有成为潜在生物防治剂的潜力。