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A novel panel of yeast assays for the assessment of thiamin and its biosynthetic intermediates in plant tissues

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Title: A novel panel of yeast assays for the assessment of thiamin and its biosynthetic intermediates in plant tissues
Authors: Strobbe, Simon, Verstraete, Jana, Fitzpatrick, Thérésa Bridget, Faustino, Maria, Lourenço, Tiago F, Oliveira, M Margarida, Stove, Christophe, Van Der Straeten, Dominique
Source: New Phytol
NEW PHYTOLOGIST
Publisher Information: Wiley, 2022.
Publication Year: 2022
Subject Terms: Nutritional improvement, 0301 basic medicine, Thiamine / metabolism, Thiamine / chemistry, Arabidopsis, PYROPHOSPHOKINASE, turbidimetry, GENE FAMILY, REDUNDANCY, Saccharomyces cerevisiae, METABOLISM, Vitamin quantification, biofortification, SACCHAROMYCES-CEREVISIAE, 03 medical and health sciences, microbiological assays, Tandem Mass Spectrometry, Saccharomyces cerevisiae / metabolism, STRESS TOLERANCE, Thiamine, Arabidopsis / metabolism, 2. Zero hunger, 0303 health sciences, vitamin quantification, Research, Biology and Life Sciences, RIBOSWITCH, Microbiological assays, Tandem Mass Spectrometry / methods, nutritional improvement, BIOFORTIFICATION, ARABIDOPSIS-THALIANA, VITAMIN-B6 BIOSYNTHESIS, metabolic engineering, Biofortification, Metabolic engineering, Turbidimetry, Chromatography, Liquid
Description: Summary Thiamin (or thiamine), known as vitamin B1, represents an indispensable component of human diets, being pivotal in energy metabolism. Thiamin research depends on adequate vitamin quantification in plant tissues. A recently developed quantitative liquid chromatography–tandem mass spectrometry (LC–MS/MS) method is able to assess the level of thiamin, its phosphorylated entities and its biosynthetic intermediates in the model plant Arabidopsis thaliana, as well as in rice. However, their implementation requires expensive equipment and substantial technical expertise. Microbiological assays can be useful in deter‐mining metabolite levels in plant material and provide an affordable alternative to MS‐based analysis. Here, we evaluate, by comparison to the LC–MS/MS reference method, the potential of a carefully chosen panel of yeast assays to estimate levels of total vitamin B1, as well as its biosynthetic intermediates pyrimidine and thiazole in Arabidopsis samples. The examined panel of Saccharomyces cerevisiae mutants was, when implemented in microbiological assays, capable of correctly assigning a series of wild‐type and thiamin biofortified Arabidopsis plant samples. The assays provide a readily applicable method allowing rapid screening of vitamin B1 (and its biosynthetic intermediates) content in plant material, which is particularly useful in metabolic engineering approaches and in germplasm screening across or within species.
Document Type: Article
Other literature type
File Description: application/pdf
Language: English
ISSN: 1469-8137
0028-646X
DOI: 10.1111/nph.17974
Access URL: https://onlinelibrary.wiley.com/doi/pdfdirect/10.1111/nph.17974
https://pubmed.ncbi.nlm.nih.gov/35037254
https://archive-ouverte.unige.ch/unige:168730
https://doi.org/10.1111/nph.17974
https://biblio.ugent.be/publication/8735809
https://biblio.ugent.be/publication/8735809/file/8743091
http://doi.org/10.1111/nph.17974
http://hdl.handle.net/1854/LU-8735809
Rights: CC BY
Accession Number: edsair.doi.dedup.....829c31c74d6b289e75900b27738bded9
Database: OpenAIRE
Description
Abstract:Summary Thiamin (or thiamine), known as vitamin B1, represents an indispensable component of human diets, being pivotal in energy metabolism. Thiamin research depends on adequate vitamin quantification in plant tissues. A recently developed quantitative liquid chromatography–tandem mass spectrometry (LC–MS/MS) method is able to assess the level of thiamin, its phosphorylated entities and its biosynthetic intermediates in the model plant Arabidopsis thaliana, as well as in rice. However, their implementation requires expensive equipment and substantial technical expertise. Microbiological assays can be useful in deter‐mining metabolite levels in plant material and provide an affordable alternative to MS‐based analysis. Here, we evaluate, by comparison to the LC–MS/MS reference method, the potential of a carefully chosen panel of yeast assays to estimate levels of total vitamin B1, as well as its biosynthetic intermediates pyrimidine and thiazole in Arabidopsis samples. The examined panel of Saccharomyces cerevisiae mutants was, when implemented in microbiological assays, capable of correctly assigning a series of wild‐type and thiamin biofortified Arabidopsis plant samples. The assays provide a readily applicable method allowing rapid screening of vitamin B1 (and its biosynthetic intermediates) content in plant material, which is particularly useful in metabolic engineering approaches and in germplasm screening across or within species.
ISSN:14698137
0028646X
DOI:10.1111/nph.17974