多孔板-mtt比色法评价植物和微生物代谢产物的抗真菌活性 |
|
|
多孔板-MTT比色法评价植物和微生物代谢产物的抗真菌活性 Evaluation of antifungal activity of plant and microbial metabolites by microplate-MTT colorimetric assay 发布时间：2011-06-30 浏览量：59 收藏数：0 评论数：0 总览 评价 钟灵允1， 单体江2， 刘浩2， 赵江林2， 徐梁2， 周立刚2* （ 1、中国农业大学农学与生物技术学院； 2、中国农业大学农学与生物技术学院，北京 100193； ）
摘要： 多孔板-MTT比色法测定植物和微生物代谢产物对真菌抑制活性的步骤为：在多孔板的每孔中依次加入浓度为100000 孢子/mL 的供试真菌孢子悬液90 μL，不同浓度的药液10 μL。25 oC暗培养48 h，然后每孔中加入8 mg/mL 的MTT 溶液10 μL，继续培养10 h 后，离心去上清，加入DMSO 150 μL，振荡30 min，离心后上清液在510 nm 测定吸光值。采用上述条件测定了白屈菜红碱对稻瘟病菌和西瓜枯萎病菌的MIC值分别为80 μg/mL和1.5 μg/mL，IC50值分别为21.99 μg/mL和0.78 μg/mL；Diepoxin ζ对稻瘟病菌的MIC和IC50值分别为200 μg/mL和96.21 μg/mL。多孔板-MTT比色法为快速有效地筛选和评价植物和微生物抗真菌活性成分创造了条件。
关键词： 多孔板-MTT比色法；植物代谢产物；微生物代谢产物；抗真菌活性 ZHONG Lingyun1， SHAN Tijiang2， LIU Hao2， ZHAO Jianglin2， XU Liang2， ZHOU Ligang2* （ 1、College of Agronomy and Biotechnology, China Agricultural University； 2、College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China； ）
Abstract： An optimal experimental procedure with the microplate-MTT colorimetric assay was obtained, that was: 1) addition of 90 μL of fungal spore suspension at concentration of 100000 spores/mL in each well of the 96-well microplate; 2) then addition of 10 μL of the sample solution with its different concentration; 3) cultivation for 48 h at 25 oC in darkness; 4) 10 μL of MTT solution at 8 mg/mL was added into each well; 5) after incubation for another 10 h at 25 oC, centrifuged and the supernatant was removed, 150 μL of DMSO was added; 6) after shaking for another 30 min at 15 rpm, light absorption values at 510 nm were obtained for the supernatant transferred in a new plate with a microplate spectrophotometer. With the optimal MTT assay mentioned above, the minimum inhibitory concentration (MIC) values of chelerythrine from Macleaya cordata on Fusarium oxysporum f.sp. niveum and Magnaporthe oryzae were 80 μg/mL and 1.5 μg/mL respectively, and the median inhibitory concentration (IC50) values were 21.99 μg/mL and 0.78 μg/mL, correspondingly. The MIC and IC50 values of diepoxin ζ from the endophytic fungus Dzf12 on Magnaporthe oryzae were 200 μg/mL and 96.21 μg/mL. It indicated that the antifungal metabolites from plants and microbes could be evaluated fast and micro-quantitatively by using microplate-MTT colorimetric assay that is favorable for screening antifungal components with a high throughout.
Keywords： microplate-MTT colorimetric assay; plant metabolites; microbial metabolites; antifungal activity
多孔板-mtt比色法评价植物和微生物代谢产物的抗真菌活性 来自: 免费论文网www.paper800.com
远程下载:《多孔板-mtt比色法评价植物和微生物代谢产物的抗真菌活性》论文(PDF全文)
|
|
|
|
|
|
|
