MTT比色法测定烟酰胺类化合物对植物病原细菌的抑菌活性
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摘要
【目的】探究MTT比色法用于合成化合物对植物病原细菌抑菌活性筛选的可行性。【方法】将7个烟酰胺类化合物分别用MTT比色法和平板扩散法对番茄疮痂病菌(Xanthomonas vesicatoria)、烟草青枯病菌(Ralstonia solanacearum)和丁香假单胞杆菌(Pseudomonas syringae)的抑菌活性进行筛选并测定其有效中浓度EC50。【结果】采用平板扩散法进行抑菌活性初筛时,未能筛选出有活性的化合物;采用MTT比色法筛选出N-(3-三氟甲基)-苯基烟酰胺具有抑菌活性,其对番茄疮痂病菌(X. vesicatoria)、烟草青枯病菌(R. solanacearum)和丁香假单胞杆菌(P. syringae)的EC50分别为3.87、2.05、10.33 mg/mL。【结论】MTT比色法能更加准确地评价脂溶性化合物对细菌的抑菌活性,并具有广泛的应用前景。
[Purpose] To investigate the possibility of antibacterial activity of 7 synthetic nicotinamide compounds against 3 kinds of pathogenic bacteria by MTT assay.[Methods]We evaluated bactericidal activity of 7 synthetic nicotinamide compounds against Xanthomonas vesicatoria, Ralstonia solanacearum and Pseudomonas syringae by MTT assay and plate diffusion method. MTT assay performed to identify bactericidal activities of N-(3-(trifluoromethyl) phenyl) nicotinamide.[Result]The EC50 of N-(3-(trifluoromethyl) pheny) nicotinamide against X. vesicatoria, R. solanacearum and P. syringae by MTT assay were 3.87, 2.05 and 10.33 mg/mL.[Conclusion]The results indicated that MTT assay was more effective than plate diffusion method in evaluating bactericidal activity, and has higher value on application.
[Purpose] To investigate the possibility of antibacterial activity of 7 synthetic nicotinamide compounds against 3 kinds of pathogenic bacteria by MTT assay.[Methods]We evaluated bactericidal activity of 7 synthetic nicotinamide compounds against Xanthomonas vesicatoria, Ralstonia solanacearum and Pseudomonas syringae by MTT assay and plate diffusion method. MTT assay performed to identify bactericidal activities of N-(3-(trifluoromethyl) phenyl) nicotinamide.[Result]The EC50 of N-(3-(trifluoromethyl) pheny) nicotinamide against X. vesicatoria, R. solanacearum and P. syringae by MTT assay were 3.87, 2.05 and 10.33 mg/mL.[Conclusion]The results indicated that MTT assay was more effective than plate diffusion method in evaluating bactericidal activity, and has higher value on application.
引文
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[14]HAMID R,ROTSHTEYN Y,RABADI L,et al.Comparison of alamar blue and MTT assays for high throughput screening[J].Toxicology in Vitro An International Journal Published in Association with Bibra,2004,18(5):703.DOI:10.1016/j.tiv.2004.03.012.
[15]吕献海.基于蛋白靶标新型多取代吡唑衍生物的设计合成及生物活性评价[D].合肥:安徽农业大学,2017.
[16]杜士杰,覃兆海.啶酰菌胺类似物的合成及抑菌活性[J].农药学学报,2016,18(4):424.DOI:10.16801/j.issn.1008-7303.2016.0059.
[17]王凯博,普特,罗艳,等.烟酰胺类化合物的合成及其抑菌活性[J].云南农业大学学报(自然科学),2018,33(2):218.DOI:10.12101/j.issn.1004-390X(n).20170403.
[2]陈亮,司乃国.植物细菌性病害与化学防治[C]//第六届中国植物病害化学防治学术研讨会论文集,2010.
[3]陈亮,张力群,刘君丽,等.杀细菌剂高通量筛选研究[J].农药,2015,54(9):678.DOI:10.16820/j.cnki.1006-0413.2015.09.016.
[4]于志同,韩立荣,吴华,等.34种植物丙酮提取物抑菌活性的初步筛选[J].西北农林科技大学学报(自然科学版),2012,40(9):72.DOI:10.13207/j.cnki.jnwafu.2012.09.016.
[5]刘敬,王琼,赵斌.天然抑菌剂的提取工艺及抗芒果病原菌作用研究[J].安徽农业科学,2011,39(13):7729.DOI:10.13989/j.cnki.0517-6611.2011.13.188.
[6]李树正,张素华.杀菌剂筛选方法的研究[J].农药,1997(9):20.DOI:10.16820/j.cnki.1006-0413.1997.09.004.
[7]李上标,裴淑艳,蒋超,等.MTT比色法研究应用进展[J].西北民族大学学报(自然科学版),2013,34(3):68.DOI:10.14084/j.cnki.cn62-1188/n.2013.03.014.
[8]MOSMANN T.Rapid colorimetric assay for cellular growth and survival:application to proliferation and cytotoxicity assays[J].Journal of Immunological Methods,1983,65(1/2):55.DOI:10.1016/0022-1759(83)90303-4.
[9]薛雅蓉,庄重,钟昭昭,等.MTT比色法在细胞生物学实验教学中的实验安排与效果分析[J].高校生物学教学研究(电子版),2015,5(3):45.DOI:10.3868/j.issn2095-1574.2015.01.020.
[10]ABE K,MATSUKI N.Measurement of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)reduction activity and lactate dehydrogenase release using MTT[J].Neuroscience Research,2000,38(4):325.DOI:10.1016/S0168-0102(00)00188-7.
[11]郭以河,张闽峰,孟家榕.MTT法体外药敏试验研究进展及临床应用[J].东南国防医药,2005,7(1):77.DOI:10.3969/j.issn.1672-271X.2005.01.044.
[12]唐静,谈满良,赵江林,等.多孔板-MTT比色法测定植物抗菌成分对细菌的抑制活性[J].天然产物研究与开发,2008,20(6):949.DOI:10.16333/j.1001-6880.2008.06.001.
[13]黄银久,宋宝安,金林红,等.MTT比色法抗肿瘤药物筛选实验条件和数据优化探索[J].激光生物学报,2009,18(3):401.DOI:10.3969/j.issn.1007-7146.2009.03.021.
[14]HAMID R,ROTSHTEYN Y,RABADI L,et al.Comparison of alamar blue and MTT assays for high throughput screening[J].Toxicology in Vitro An International Journal Published in Association with Bibra,2004,18(5):703.DOI:10.1016/j.tiv.2004.03.012.
[15]吕献海.基于蛋白靶标新型多取代吡唑衍生物的设计合成及生物活性评价[D].合肥:安徽农业大学,2017.
[16]杜士杰,覃兆海.啶酰菌胺类似物的合成及抑菌活性[J].农药学学报,2016,18(4):424.DOI:10.16801/j.issn.1008-7303.2016.0059.
[17]王凯博,普特,罗艳,等.烟酰胺类化合物的合成及其抑菌活性[J].云南农业大学学报(自然科学),2018,33(2):218.DOI:10.12101/j.issn.1004-390X(n).20170403.
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