Defez Roberto, Andreozzi Anna, Bianco Carmen
Institute of Biosciences and BioResources, Via P. Castellino 111, Naples, Italy.
Bio Protoc. 2017 Jan 20;7(2):e2115. doi: 10.21769/BioProtoc.2115.
This protocol describes the use of the 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) salt to evaluate the cell redox potential of rhizobia cells. The production of brightly colored and insoluble 1,3,5-Triphenyltetrazolium formazan arising from TTC reduction is irreversible and can be easily quantified using a spectrophotometer. This protocol allows the production of reproducible results in a relatively short time for 1021 cells grown both in exponential and stationary phases. The results here presented show that the cells deriving from exponential-phase cultures had increased cell redox potential as compared to the ones deriving from stationary-phase cultures. This means that under exponential growth conditions the cells generate higher amount of reducing equivalents needed for TTC reduction.
本方案描述了使用2,3,5-三苯基-2H-氯化四氮唑（TTC）盐来评估根瘤菌细胞的细胞氧化还原电位。TTC还原产生的颜色鲜艳且不溶性的1,3,5-三苯基甲臜是不可逆的，并且可以使用分光光度计轻松定量。该方案能够在相对较短的时间内，对处于指数生长期和稳定期的1021个细胞产生可重复的结果。此处呈现的结果表明，与来自稳定期培养物的细胞相比，来自指数期培养物的细胞具有更高的细胞氧化还原电位。这意味着在指数生长条件下，细胞产生更多用于TTC还原所需的还原当量。
