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摘要：紫苏醛是紫苏中的活性成分，是天然紫苏油的特征香气成分，存在于天然紫苏油中。本实验以紫苏醛为研究对象，通过细胞膜损伤实验探究紫苏醛对细胞膜的作用，以线粒体为抗真菌作用靶标，研究紫苏醛对黄曲霉和黑曲霉的抗真菌作用，主要通过两个方面研究：根据荧光染料Rh123观察紫苏醛对黑曲霉、黄曲霉膜电位（MMP）的变化影响；根据荧光探针DCFH-DA观察紫苏醛对黑曲霉、黄曲霉线粒体活性氧（ROS）的变化影响。结果表明，随着紫苏醛浓度增高，PI染色的黄曲霉细胞数从1.1%增加到96.0%，PI染色的黑曲霉细胞数从10.8%增加到61.3%；Rh123染色的黄曲霉细胞数目从2.2%增加到92.4%，Rh123染色的黑曲霉细胞数目从0.2%增加到87.1%；利用DCFH-DA荧光探针黄曲霉的染色数目从2.6%增加到45.7%,黑曲霉的染色数目从0.6%增加到67.0%，紫苏醛浓度的越高，黄曲霉和黑曲霉中被PI染色的细胞数目越多，荧光强度增加，MMP增加，线粒体ROS的积累越多，紫苏醛对黄曲霉和黑曲霉具有抑菌作用，抑制效果与浓度呈正相关。33496
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毕业论文关键词： 紫苏；紫苏醛；黑曲霉；黄曲霉；线粒体
Perillaldehyde effect on Aspergillus niger and
(School of Life Sciences, Jiangsu Normal University, Xuzhou,Jiangsu,221116,China)
Abstract: Perillaldehyde in perilla extract, is the active ingredient in perilla, is characteristic of natural perilla oil aroma components, existing natural perilla oil. This experiment by perillaldehyde as the research object, through the cell membrane damage experiment to explore the effect of perillaldehyde on cell membrane, mitochondria as antifungal effect targets, study of perillaldehyde of A. niger and A. flavus antifungal effect, mainly through two aspects of research: According to the fluorescence dye Rh123 observation perillaldehyde of aspergillus Niger, aspergillus flavus MMP change; according to the fluorescence probe DCFH-DA observation perillaldehyde of A. niger, A. flavus mitochondrial ROS changes . The results show that with perillaldehyde concentration increased, PI staining of A. flavus cells was increased from 1.1% to 96.0%, PI staining of A. niger cells was increased from 10.8% to 10.8%; Rh123 dyeing of A. flavus cell number increased from 2.2% to 92.4%, the number of A. niger cells Rh123 dyeing increased from 0.2% to 87.1%; Using DCFH-DA fluorescent probe dyeing number increased from 2.6% to 45.7% of A. flavus, A. niger dyeing number increased from 0.6% to 67.0%, the higher the concentration of perillaldehyde, A. flavus and A. niger in the number of cells stained by PI, the fluorescence intensity increases, MMP increased, the more accumulation of mitochondrial ROS, perillaldehyde to A. flavus, A. niger has bacteriostatic action, inhibition effect was positively correlated with concentration.
Key words: Perilla; Perillaldehyde; A. niger; A. flavus; Mitochondria.
目 录
前言 1
1 材料与方法 2
1.1供试材料 2
1.1.1 原料 2
1.1.2 实验试剂 3
1.1.3 试剂的制配 3
1.1.4 培养基 4
1.1.5 实验仪器 4
1.2 方法 5
1.2.1 菌种活化 5
1.2.2 孢子悬浮液的制备 5
1.2.3 细胞膜损伤 5
1.2.4 线粒体膜电位(MMP)的测定 5
1.2.5 线粒体活性氧(ROS)的测定 6
2 结果与分析 7
2.1 黄曲霉细胞膜损伤的测定 7
2.2 黑曲霉细胞膜损伤的测定 9
2.3 黄曲霉MMP的测定 11
2.4 黑曲霉MMP的测定 13
2.5 黄曲霉线粒体ROS的测定 15