Metabolic reprogramming in finger millet following ethyl methanesulfonate mutagenesis.
5 affiliations
- 1Department of Agriculture and Animal Health, University of South Africa, Florida Park, Roodepoort, 1710, South Africa. [email protected].
- 2Department of Agriculture and Animal Health, University of South Africa, Florida Park, Roodepoort, 1710, South Africa.
- 3Center of Excellence in Mass Spectrometry for Southern Africa (CEMMSA), University of Venda, University Rd, Thohoyandou, 0950, South Africa.
- 4Agricultural Research Council - Grain Crops Institute, Potchefstroom, South Africa.
- 5Department of Plant and Soil Sciences, Faculty of Science, Engineering and Agriculture, University of Venda, Thohoyandou, South Africa.
Abstract
Background
Finger millet is a climate-resilient cereal crop with strong potential for improving food security in marginal environments; however, its improvement has been constrained by low natural genetic variability. This limitation has prompted the use of mutation breeding to expand genetic diversity in underutilized crops. Chemical mutagenesis, particularly using ethyl methanesulfonate (EMS) has been widely applied for this purpose, yet its concentration-dependent metabolic effects during plant development remain insufficiently understood. This study aimed to characterize metabolic changes in seedlings of four finger millet genotypes subjected to 0% (control), 0.10%, 0.40%, and 0.70% EMS using untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomics. Seeds were treated for one hour and grown in a randomized complete design with three replicates per treatment. Aboveground tissues were harvested six weeks after planting, and metabolic profiles were compared across treatments.Results
Principal component analysis revealed clear concentration-dependent separation, with PC1 and PC2 explaining 63.5% of the variance. Control samples formed a compact cluster, while 0.10% EMS-treated seedlings showed slight but distinct separation along PC2. Supervised models (PLS-DA, sPLS-DA, and OPLS-DA) confirmed strong discrimination among treatments and identified key metabolites driving separation. A total of 2 262 metabolic features were detected, of which 400 were putatively annotated at MSI level 2 based on accurate mass and MS/MS spectral matching. Differential analysis identified 81, 31, and 50 significantly altered metabolites at 0.10%, 0.40% and 0.70% EMS concentrations, respectively, spanning organic acids, phenolics, flavonoids, lipids, alkaloids, amino acids and terpenoids. The lowest concentration (0.10%) induced the broadest response, marked by increased phenolic acids and flavonoids, alongside mixed regulation of organic acids, including elevated malic and succinic acids and reduced oxaloacetic acid. However, 0.40% of EMS resulted in widespread metabolic repression, with mesaconic acid among the few consistently upregulated metabolites. The highest concentration (0.70%) triggered extensive metabolic remodeling, including perturbation of tricarboxylic acid cycle intermediates and activation of secondary metabolites including chlorogenic acid. Pairwise comparisons revealed structured metabolic transitions, including suppression of phenylpropanoid pathways and shifts in central carbon metabolism with increasing EMS levels. Heatmap analysis highlighted genotype and replicate-specific responses, indicating heterogeneity in metabolic adaptation. Certain metabolites, including 2-caffeoyl-L-tartaric acid, showed strong accumulation under moderate and high EMS treatments, suggesting localized metabolic plasticity.Conclusion
These findings demonstrate that EMS induces concentration-dependent and genotype-specific metabolic reprogramming in finger millet, providing insight into early biochemical responses to mutagenesis and supporting the application of metabolomics in optimizing mutation breeding strategies.
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BibTeX
@article{mutanda2026metabolic,
title = {{Metabolic reprogramming in finger millet following ethyl methanesulfonate mutagenesis.}},
author = {Maltase Mutanda and Sandiswa Figlan and Ntakadzeni E Madala and Nemera G Shargie and Eastonce T Gwata},
journal = {BMC genomics},
year = {2026},
month = aug,
doi = {10.1186/s12864-026-13257-x},
url = {https://europepmc.org/article/MED/42967986},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Metabolic reprogramming in finger millet following ethyl methanesulfonate mutagenesis.
AU - Maltase Mutanda
AU - Sandiswa Figlan
AU - Ntakadzeni E Madala
AU - Nemera G Shargie
AU - Eastonce T Gwata
PY - 2026
DA - 2026/08/01
JO - BMC genomics
DO - 10.1186/s12864-026-13257-x
UR - https://europepmc.org/article/MED/42967986
AB - Background Finger millet is a climate-resilient cereal crop with strong potential for improving food security in marginal environments; however, its improvement has been constrained by low natural genetic variability. This limitation has prompted the use of mutation breeding to expand genetic diversity in underutilized crops. Chemical mutagenesis, particularly using ethyl methanesulfonate (EMS) has been widely applied for this purpose, yet its concentration-dependent metabolic effects during plant development remain insufficiently understood. This study aimed to characterize metabolic changes in seedlings of four finger millet genotypes subjected to 0% (control), 0.10%, 0.40%, and 0.70% EMS using untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomics. Seeds were treated for one hour and grown in a randomized complete design with three replicates per treatment. Aboveground tissues were harvested six weeks after planting, and metabolic profiles were compared across treatments.Results Principal component analysis revealed clear concentration-dependent separation, with PC1 and PC2 explaining 63.5% of the variance. Control samples formed a compact cluster, while 0.10% EMS-treated seedlings showed slight but distinct separation along PC2. Supervised models (PLS-DA, sPLS-DA, and OPLS-DA) confirmed strong discrimination among treatments and identified key metabolites driving separation. A total of 2 262 metabolic features were detected, of which 400 were putatively annotated at MSI level 2 based on accurate mass and MS/MS spectral matching. Differential analysis identified 81, 31, and 50 significantly altered metabolites at 0.10%, 0.40% and 0.70% EMS concentrations, respectively, spanning organic acids, phenolics, flavonoids, lipids, alkaloids, amino acids and terpenoids. The lowest concentration (0.10%) induced the broadest response, marked by increased phenolic acids and flavonoids, alongside mixed regulation of organic acids, including elevated malic and succinic acids and reduced oxaloacetic acid. However, 0.40% of EMS resulted in widespread metabolic repression, with mesaconic acid among the few consistently upregulated metabolites. The highest concentration (0.70%) triggered extensive metabolic remodeling, including perturbation of tricarboxylic acid cycle intermediates and activation of secondary metabolites including chlorogenic acid. Pairwise comparisons revealed structured metabolic transitions, including suppression of phenylpropanoid pathways and shifts in central carbon metabolism with increasing EMS levels. Heatmap analysis highlighted genotype and replicate-specific responses, indicating heterogeneity in metabolic adaptation. Certain metabolites, including 2-caffeoyl-L-tartaric acid, showed strong accumulation under moderate and high EMS treatments, suggesting localized metabolic plasticity.Conclusion These findings demonstrate that EMS induces concentration-dependent and genotype-specific metabolic reprogramming in finger millet, providing insight into early biochemical responses to mutagenesis and supporting the application of metabolomics in optimizing mutation breeding strategies.
SN - 1471-2164
ER -