Simple and semi-high throughput determination of total phenolic, anthocyanin, flavonoid content, and total antioxidant capacity of model and crop plants for cell physiological phenotyping

Abstract

Plants biosynthesize a wide range of antioxidants capable of attenuating ROS-induced oxidative damage. There exist several in vitro methods to analyze antioxidants and total antioxidant capacity from different tissues and of various plant species. We have established a single, fast and cost-efficient extraction protocol combined with a semihigh throughput 96-well plate assay methods for determination of the level of the key antioxidants phenolics, anthocyanins and flavonoids in combination with the determination of total antioxidant capacity using ferric reducing antioxidant power (FRAP) and trolox equivalent antioxidant capacity (TEAC). The method was optimized and verified with samples from different strawberry species and cultivars with known differences in the parameters measured. This method proved to be suitable for analyses of eight model and crop plants, and distinct antioxidant signatures were determined for the different tissues and organs analyzed, including leaf, root, fruit, spike, and tuber samples. The method was robust and was shown in two case studies to be a resource-efficient and fast experimental platform also to assess biotic and abiotic stress responses, notably including fungal infection and the impact of a progressive drought regime. Since method was adapted for a semi-high throughput 96-well assay format it is well-suited for integration of cell physiological phenotyping into a holistic phenomics approach for germplasm assessment and plant breeding screening. This analytical platform uses microplate spectrophotometer which proved to be suitable to determine the antioxidant contents and total antioxidant capacity signatures of various plant species and tissues with similar findings as reported in literature.

Keywords: Anthocyanin; Antioxidant Capacity; FRAP; Flavonoid; Phenolic; TEAC.