天然产物研究与开发2012，Vol.24Issue(1)：20-24,5.
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
摘要
Abstract
An optimal experimental procedure with the microplate-MTT colorimetric assay was developed,that was as follows :90 μL of fungal spore suspension at concentration of 105 spores/mL was added in each well of the 96-well micro-plate first; 10 μL of the sample solution with its different concentration was then added,and cultivated for 48 h at 25 X, in darkness; 10 μL of MTT solution at 8 mg/mL was added into each well,and incubated for another 10 h at 25 °C and centrifuged ;The supernatant was removed, and 150 μL of DMSO was added; 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 spectro-photometer. With the optimal MTT assay mentioned above,the minimum inhibitory concentration (MIC) values of chel-erythrine from Macleaya cordata on Fusarium oxysporum f. sp. Niveum and Magnaporthe oryzae were 80 and 1.5 μg/mL respectively,and the median inhibitory concentration (IC50) values were 21.99 and 0.78 μg/mL,correspondingly. The MIC and IC50 values of diepoxin ξ from the endophytic fungus Dzfl2 on Magnaporthe oryzae were 200 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.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Key words
microplate-MTT colorimetric assay/ plant metabolites/ microbial metabolites/ antifungal activity分类
农业科技引用本文复制引用
钟灵允,王兰,单体江,刘浩,赵江林,徐梁,周立刚..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[J].天然产物研究与开发,2012,24(1):20-24,5.基金项目
高等学校博士学科点专项科研基金(20060019048) （20060019048）
新疆生产建设兵团塔里木盆地生物资源保护利用重点实验室开放基金(BRZD1102) （BRZD1102）
国家自然科学基金(31071710) （31071710）
