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A critical comparison of two high-throughput ascorbate analyses methods for plant samples

机译：两种植物样品高通量抗坏血酸分析方法的关键比较

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摘要

Ascorbate (AsA) is an important metabolite involved in stress response and development of plants. Therefore it is necessary to quantify the AsA content in many fields of plant science, including high throughput and critical applications. In this study we compared two different microplate-based AsA assays, which are suitable for high throughput applications: an ascorbate oxidase (AO)-based assay and a dipyridyl (DPD)-based assay. These methods were compared in critical applications, i.e. (i) when AsA concentrations were very low such as in apoplastic extracts, (ii) when plants contained pigments interfering with the spectrometric measurements, and (iii) when plants contained high iron concentration interfering with the color reactions. The precision of measurements was higher with the DPD method, as illustrated by higher recovery rates of internal AsA standards. On the other hand, the AO method was more sensitive to low levels of AsA. This was an advantage in determining apoplastic AsA concentration in rice, which was substantially lower than that of whole tissues. The AO method also had the advantage that plant pigments and high iron concentrations in plants tissues did not interfere with the analysis, as opposed to the DPD assay. In conclusion, both assays had advantages and the choice of a suitable method depends on the specific application.
机译：抗坏血酸（AsAb）是一种重要的代谢产物，参与植物的应激反应和发育。因此，有必要在植物科学的许多领域中量化AsA含量，包括高通量和关键应用。在本研究中，我们比较了两种适用于高通量应用的基于微孔板的AsA分析：基于抗坏血酸氧化酶（AO）的分析和基于二吡啶基（DPD）的分析。在关键应用中比较了这些方法，即（i）当AsA浓度非常低时，例如在质外塑性提取物中;（ii）当植物中含有干扰光谱测定的色素时;（iii）当植物中高铁浓度干扰了光谱分析时颜色反应。 DPD方法的测量精度更高，内部AsA标准品的回收率更高。另一方面，AO方法对低水平的AsA更为敏感。这是测定水稻中质外体AsA浓度的一个优势，该浓度显着低于整个组织。相对于DPD分析，AO方法还具有植物色素和植物组织中高铁浓度不会干扰分析的优势。总之，两种测定法均具有优势，合适方法的选择取决于具体应用。

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