赵妗颐, 肖洋, 李祝, 张钦语, 杨佩斯. 黑曲霉xj粗提物的拮抗机制及其抗氧化活性[J]. 农药学学报, 2020, 22(4): 642-651. DOI: 10.16801/j.issn.1008-7303.2020.0084
    引用本文: 赵妗颐, 肖洋, 李祝, 张钦语, 杨佩斯. 黑曲霉xj粗提物的拮抗机制及其抗氧化活性[J]. 农药学学报, 2020, 22(4): 642-651. DOI: 10.16801/j.issn.1008-7303.2020.0084
    ZHAO Jinyi, XIAO Yang, LI Zhu, ZHANG Qinyu, YANG Peisi. Antagonistic mechanism and antioxidant of crude extract of Aspergillus niger xj[J]. Chinese Journal of Pesticide Science, 2020, 22(4): 642-651. DOI: 10.16801/j.issn.1008-7303.2020.0084
    Citation: ZHAO Jinyi, XIAO Yang, LI Zhu, ZHANG Qinyu, YANG Peisi. Antagonistic mechanism and antioxidant of crude extract of Aspergillus niger xj[J]. Chinese Journal of Pesticide Science, 2020, 22(4): 642-651. DOI: 10.16801/j.issn.1008-7303.2020.0084

    黑曲霉xj粗提物的拮抗机制及其抗氧化活性

    Antagonistic mechanism and antioxidant of crude extract of Aspergillus niger xj

    • 摘要: 为探究黑曲霉Aspergillus niger xj 菌株粗提物SE对魔芋白绢病病原菌——齐整小核菌Sclerotium rolfsii R-67的拮抗机制及其抗氧化活性,采用菌丝生长速率法,研究了SE对R-67的抑制作用,进而通过测定处理后菌株的电导率、核酸蛋白泄露、三羧酸循环关键酶琥珀酸脱氢酶 (SDH) 及苹果酸脱氢酶 (MDH)比活力、胞内活性氧 (ROS) 水平及核酸含量,对SE的拮抗机制进行了研究,并通过1,1-二苯基-2-三硝基苯肼 (DPPH) 法及2,2-联氮基-双-(3-乙基苯并噻唑啉-6-磺酸) 二铵盐 (ABTS) 法测定了其抗氧化活性。通过气相色谱-质谱联用 (GC-MS)方法,从粗提物SE中共检测到12种化合物,其中主要成分为5-羟甲基糠醛(质量分数 97.96%)。生物试验结果表明:SE能够抑制R-67菌丝的生长,其IC50值为0.46 mg/mL;SE处理可导致R-67菌液的电导率升高及核酸蛋白含量增加,抑制其三羧酸循环中关键酶SDH与MDH 的活力,增加菌体细胞内ROS含量,使细胞核4',6-二脒基-2-苯基吲哚 (DAPI) 染色的荧光强度减弱,据此初步推测其作用机制是通过影响R-67菌体细胞膜完整性、损伤细胞结构、抑制三羧酸循环及核酸合成等途径而发挥抑菌作用。研究表明,黑曲霉xj粗提物SE在魔芋白绢病的生物防治中具有良好的应用前景,此外SE对DPPH及ABTS自由基有一定的清除能力,有望应用于抗氧化功能性产品中。

       

      Abstract: The aim of this experiment was to investigate the antioxidant activity and antagonistic mechanism of crude extract SE of Aspergillus niger xj against Sclerotium rolfsii R-67 in Amorphophallus konjac K. Koch. The inhibition of SE on R-67 was studied by the growth rate method. Then the bacteriostatic mechanism of SE was studied by measuring the conductivity, nucleic acid and protein leakage, the specific activity of tricarboxylic acid cycle key enzymes (succinate dehydrogenase (SDH) and malate dehydrogenation (MDH)), the level of intracellular reactive oxygen species (ROS) and nucleic acid content. The antioxidant activity was determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods. By gas chromatography-mass spectrometry (GC-MS) method, a total of 12 compounds were detected from the crude extract SE, of which the main component was 5-hydroxymethylfurfural (mass fraction 97.96%).   Biological test results showed that SE could inhibit the growth of R-67, and the IC50 value was 0.46 mg/mL. SE could increase the conductivity and nucleic acid protein content of R-67, inhibit the specific activity of key enzymes in the tricarboxylic acid cycle SDH to MDH, increase the intracellular ROS content, and weaken the fluorescence intensity of (4′,6-diamidine-2-phenylindole) DAPI staining. Furthermore, it was preliminarily speculated that the mechanism of action included the inhibition of the cell membrane integrity of R-67, the damage of the cell structure, and the inhibition of the tricarboxylic acid cycle. The results demonstrated that the crude extraction of A. niger xj had a good application prospect in the biological control of A. konjac K. Koch during storage. And SE had a moderate ability to remove DPPH and ABTS free radicals, which was promising to be used in antioxidant functional products.

       

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