en
Scientific article
Open access
English

A protocol for a turbidimetric assay using a Saccharomyces cerevisiae thiamin biosynthesis mutant to estimate total vitamin B1 content in plant tissue samples

Published inPlant methods, vol. 19, no. 1
Publication date2023-12-13
First online date2023-12-13
Abstract

Understanding thiamin (thiamine; vitamin B 1 ) metabolism in plants is crucial, as it impacts plant nutritional value as well as stress tolerance. Studies aimed at elucidating novel aspects of thiamin in plants rely on adequate assessment of thiamin content. Mass spectrometry-based methods provide reliable quantification of thiamin as well as closely related biomolecules. However, these techniques require expensive equipment and expertise. Microbiological turbidimetric assays can evaluate the presence of thiamin in a given sample, only requiring low-cost, standard lab equipment. Although these microbiological assays do not reach the accuracy provided by mass spectrometry-based methods, the ease with which they can be deployed in an inexpensive and high-throughput manner, makes them a favorable method in many circumstances. However, the thiamin research field could benefit from a detailed step-by-step protocol to perform such assays as well as a further assessment of its potential and limitations.

Results

Here, we show that the Saccharomyces cerevisiae thiamin biosynthesis mutant thi6 is an ideal candidate to be implemented in a turbidimetric assay aimed at assessing the content of thiamin and its phosphorylated equivalents (total vitamer B 1 ). An optimized protocol was generated, adapted from a previously established microbiological assay using the thi4 mutant. A step-by-step guidance for this protocol is presented. Furthermore, the applicability of the assay is illustrated by assessment of different samples, including plant as well as non-plant materials. In doing so, our work provides an extension of the applicability of the microbiological assay on top of providing important considerations upon implementing the protocol.

Conclusions

An inexpensive, user-friendly protocol, including step-by-step guidance, which allows adequate estimation of vitamer B 1 content of samples, is provided. The method is well-suited to screen materials to identify altered vitamer B 1 content, such as in metabolic engineering or screening of germplasm.

eng
Keywords
  • Biofortification
  • Metabolic engineering
  • Microbiological assay
  • Nutrition
  • Nutritional value
  • Plant breeding
  • Protocol
  • Thiamine
  • Turbidimetry
  • Vitamin quantification
Citation (ISO format)
STROBBE, Simon et al. A protocol for a turbidimetric assay using a<i> Saccharomyces cerevisiae</i> thiamin biosynthesis mutant to estimate total vitamin B1 content in plant tissue samples. In: Plant methods, 2023, vol. 19, n° 1. doi: 10.1186/s13007-023-01117-8
Main files (1)
Article (Published version)
Identifiers
ISSN of the journal1746-4811
15views
2downloads

Technical informations

Creation06/05/2024 11:33:40 AM
First validation06/12/2024 1:16:32 PM
Update time06/12/2024 1:16:32 PM
Status update06/12/2024 1:16:32 PM
Last indexation06/12/2024 1:16:41 PM
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack