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评估一种简单、无毒的阿拉玛蓝细胞存活测定法以监测番茄细胞活力。

Assessment of a simple, non-toxic Alamar blue cell survival assay to monitor tomato cell viability.

作者信息

Byth H A, Mchunu B I, Dubery I A, Bornman L

机构信息

Biochemistry Division, Department of Chemistry and Biochemistry, Rand Afrikaans University, PO Box 524, Auckland Park 2006, South Africa.

出版信息

Phytochem Anal. 2001 Sep-Oct;12(5):340-6. doi: 10.1002/pca.595.

Abstract

The Alamar Blue (AB) assay, which incorporates a medox indicator that changes colour or fluorescence in response to metabolic activity, is commonly used to assess quantitatively the viability and/or proliferation of mammalian cells and micro-organisms. In this study the AB assay was adapted for the determination of the viability of plant cells. Cell suspension cultures of tomato, Lycopersicon esculentum, L., with differing viabilities, served as the experimental model for a comparison of the AB assay with the conventional 2,3,5-triphenyltetrazolium chloride (TTC) viability assay. The AB assay showed a sigmoidal relationship between cell viability and AB reduction (as quantified by spectrofluorometry or spectrophotometry), which was similar to that obtained using the TTC assay. Both assays detected a significant reduction in cell viability after 48 h exposure to virulent Ralstonia solanacearum (biovar III), while the TTC assay, in addition, revealed cell proliferation in control cells from 24 to 72 h. The TTC assay detected cell proliferation over a wider range of cell densities, while the AB assay was more rapid and versatile whilst being non-toxic and thus allowing subsequent cell analysis.

摘要

阿拉玛蓝(AB)检测法采用一种甲臜指示剂,该指示剂会根据代谢活性改变颜色或荧光,常用于定量评估哺乳动物细胞和微生物的活力及/或增殖情况。在本研究中,AB检测法被用于测定植物细胞的活力。具有不同活力的番茄(Lycopersicon esculentum L.)细胞悬浮培养物,作为将AB检测法与传统的2,3,5-三苯基氯化四氮唑(TTC)活力检测法进行比较的实验模型。AB检测法显示细胞活力与AB还原之间呈S形关系(通过荧光分光光度法或分光光度法定量),这与使用TTC检测法得到的结果相似。两种检测法均检测到在暴露于强致病力的青枯雷尔氏菌(生物变种III)48小时后细胞活力显著降低,而TTC检测法还显示对照细胞在24至72小时内有细胞增殖。TTC检测法在更广泛的细胞密度范围内检测到细胞增殖,而AB检测法更快速、用途更广,且无毒,因此可进行后续的细胞分析。

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