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摘要:橙花醇是一种植物精油，根据目前国内外的研究，植物精油具有抑菌的作用。本实验的目的是探究橙花醇抑菌机理，以黑曲霉和黄曲霉为研究对象，进一步探究橙花醇对黑曲霉和黄曲霉线粒体的作用。本试验探究在不同浓度橙花醇的影响下黑曲霉和黄曲霉细胞膜中甾醇含量的变化，来验证橙花醇抑制细胞膜中甾醇的合成。此外，还验证了在不同浓度的橙花醇影响下，对黑曲霉和黄曲霉细胞膜损伤，线粒体膜电位（MMP），活性氧的积累（ROS）的影响。实验结果表明：橙花醇对真菌的生长有一定的抑制作用，抑制效果与橙花醇的浓度呈正相关；当细胞受到挥发油的作用中时，使真菌线粒体膜电位升高，由此引起真菌线粒体中活性氧的积累，使线粒体功能受损，细胞活力降低，引起细胞死亡，从而发挥抗真菌效应。34458
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毕业论文关键词：橙花醇；细胞膜；线粒体；黑曲霉；黄曲霉；抗真菌活性
Effect of nerol on cell membrane and mitochondrial function of Aspergillus flavus and Aspergillus niger
Abstract: Nerol is a kind of plant essential oil, according to the current research at home and abroad, antimicrobial activity of plant essential oils, The purpose of this study is to explore the antibacterial mechanism of nerol, Aspergillus niger and Aspergillus flavus as the research object, to further explore the nerol of A. niger and A. flavus mitochondrial function. This experiment explore the impact of nerol in different concentrations by A. niger and A. flavus in cell membrane, verifying the synthesis of sterol neryl inhibition in cell membranes. In addition, also verifying the effects on different concentrations of Nerol in A. niger and A. flavus, damaging to the cell membrane, mitochondrial membrane potential (MMP) and The reactive oxygen species (ROS). The experimental results show that: nerol has inhibitory effect on fungal growth and the inhibitory effect was positively correlated with the concentration. When the cells were exposured to Nerol, this will result an elevation of MMP in a dose-dependent. Those caused ROS accumulation in the mitochondria, which impaired the mitochondrial function, decreased the cell viability and caused cell death, so as to achievethe the purpose of bacteriostasis.
Key words: Nerol; Cell Membrane; The mitochondria; A. niger ; A. flavus; Antifungal activity
目录
引言 1
1 材料与方法 3
1.1供试材料 3
1.1.1 原料 3
1.1.2 主要仪器设备 3
1.1.3 培养基配置 3
1.1.4 主要溶液试剂的配制 3
1.1.5 实验试剂 4
1.2实验方法 4
1.2.1 黑曲霉的接种活化 4
1.2.2 黄曲霉的接种活化 4
1.2.3 黄曲霉与黑曲霉孢子悬浮液的制备 5
1.2.4 甾醇含量的分析测定 5
1.2.5 细胞膜损伤的测定 5
1.2.6 线粒体膜电位的测定 6
1.2.7 ROS的测定 6
2 结果与分析 8
2.1 黑曲霉甾醇含量分析 8
2.2 黄曲霉甾醇含量分析 9
2.3 橙花醇对黑曲霉细胞膜损伤的分析 10
2.4 橙花醇对黄曲霉细胞膜损伤的分析 12
2.5 线粒体膜电位测定的结果分析 14
2.5.1黑曲霉线粒体膜电位结果 14
2.5.2黄曲霉线粒体膜电位结果 14
2.6.1橙花醇对黑曲霉ROS测定的影响 16
2.6.2橙花醇对黄曲霉ROS测定的影响 17
3 讨论 18
4 致谢 19