Centre for Ocean and Atmospheric Sciences, School of Environmental Sciences, University of East Anglia, Norwich, Norfolk, United Kingdom.
PLoS One. 2019 Dec 10;14(12):e0225954. doi: 10.1371/journal.pone.0225954. eCollection 2019.
The reduction of 2-para (iodophenyl)-3(nitrophenyl)-5(phenyl) tetrazolium chloride (INT) is increasingly being used as an indirect method to measure plankton respiration. Its greater sensitivity and shorter incubation time compared to the standard method of measuring the decrease in dissolved oxygen concentration, allows the determination of total and size-fractionated plankton respiration with higher precision and temporal resolution. However, there are still concerns as to the method's applicability due to the toxicity of INT and the potential differential effect of plankton cell wall composition on the diffusion of INT into the cell, and therefore on the rate of INT reduction. Working with cultures of 5 marine plankton (Thalassiosira pseudonana CCMP1080/5, Emiliania huxleyi RCC1217, Pleurochrysis carterae PLY-406, Scrippsiella sp. RCC1720 and Oxyrrhis marina CCMP1133/5) which have different cell wall compositions (silica frustule, presence/absence of calcite and cellulose plates), we demonstrate that INT does not have a toxic effect on oxygen consumption at short incubation times. There was no difference in the oxygen consumption of a culture to which INT had been added and that of a replicate culture without INT, for periods of time ranging from 1 to 7 hours. For four of the cultures (T. pseudonana CCMP1080/5, P. carterae PLY-406, E. huxleyi RCC1217, and O. marina CCMP1133/5) the log of the rates of dissolved oxygen consumption were linearly related to the log of the rates of INT reduction, and there was no significant difference between the regression lines for each culture (ANCOVA test, F = 1.696, df = 3, p = 0.18). Thus, INT reduction is not affected by the structure of the plankton cell wall and a single INT reduction to oxygen consumption conversion equation is appropriate for this range of eukaryotic plankton. These results further support the use of the INT technique as a valid proxy for marine plankton respiration.
2-对(碘代苯)-3(硝基苯)-5(苯基)四氮唑氯化物(INT)的还原越来越多地被用作测量浮游生物呼吸的间接方法。与测量溶解氧浓度下降的标准方法相比,它具有更高的灵敏度和更短的孵育时间,允许以更高的精度和时间分辨率测定总浮游生物和大小分级浮游生物呼吸。然而,由于 INT 的毒性以及浮游生物细胞壁组成对 INT 向细胞内扩散的潜在差异影响,以及对 INT 还原速率的影响,人们仍然对该方法的适用性存在担忧。我们使用 5 种海洋浮游生物(Thalassiosira pseudonana CCMP1080/5、Emiliania huxleyi RCC1217、Pleurochrysis carterae PLY-406、Scrippsiella sp. RCC1720 和 Oxyrrhis marina CCMP1133/5)的培养物进行了研究,这些浮游生物具有不同的细胞壁组成(硅质外壳、有无方解石和纤维素板),结果表明 INT 在短孵育时间内对耗氧量没有毒性作用。在添加 INT 和不添加 INT 的培养物中,在 1 至 7 小时的时间段内,耗氧量没有差异。对于四种培养物(T. pseudonana CCMP1080/5、P. carterae PLY-406、E. huxleyi RCC1217 和 O. marina CCMP1133/5),溶解氧消耗率的对数与 INT 还原率的对数呈线性关系,并且对于每种培养物的回归线之间没有显著差异(ANCOVA 检验,F = 1.696,df = 3,p = 0.18)。因此,INT 还原不受浮游生物细胞壁结构的影响,并且适用于该范围内的真核浮游生物的单一 INT 还原到耗氧量转换方程是合适的。这些结果进一步支持将 INT 技术用作海洋浮游生物呼吸的有效替代物。