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The R2R3-MYB transcription factor MtMYB134 orchestrates flavonol biosynthesis in Medicago truncatula

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Our results provide insights into the flavonol biosynthesis regulation of M. truncatula. The R2R3-MYB transcription factor MtMYB134 emerged as tool to improve the flavonol biosynthesis.

Abstract

Flavonols are plant specialized metabolites with vital roles in plant development and defense and are known as diet compound beneficial to human health. In leguminous plants, the regulatory proteins involved in flavonol biosynthesis are not well characterized. Using a homology-based approach, three R2R3-MYB transcription factor encoding genes have been identified in the Medicago truncatula reference genome sequence. The gene encoding a protein with highest similarity to known flavonol regulators, MtMYB134, was chosen for further experiments and was characterized as a functional flavonol regulator from M. truncatula. MtMYB134 expression levels are correlated with the expression of MtFLS2, encoding a key enzyme of flavonol biosynthesis, and with flavonol metabolite content. MtMYB134 was shown to activate the promoters of the A. thaliana flavonol biosynthesis genes AtCHS and AtFLS1 in Arabidopsis protoplasts in a transactivation assay and to interact with the Medicago promoters of MtCHS2 and MtFLS2 in yeast 1-hybrid assays. To ascertain the functional aspect of the identified transcription factor, we developed a sextuple mutant, which is defective in anthocyanin and flavonol biosynthesis. Ectopic expression of MtMYB134 in a multiple myb A. thaliana mutant restored flavonol biosynthesis. Furthermore, overexpression of MtMYB134 in hairy roots of M. truncatula enhanced the biosynthesis of various flavonol derivatives. Taken together, our results provide insight into the understanding of flavonol biosynthesis regulation in M. truncatula and provides MtMYB134 as tool for genetic manipulation to improve flavonol synthesis.

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Acknowledgements

We are thankful to Dr. Senjuti Sinha Roy for providing Medicago truncatula seeds. This work was supported by the core grant of National Institute of Plant Genome Research and Department of Science and Technology-SERB for Startup research grant to AP (Grant Number: SRG/2019/000503). JN and Ruchika acknowledge Council of Scientific and Industrial Research, Government of India for Senior Research Fellowships. The authors are thankful to DBT-eLibrary Consortium (DeLCON) for providing access to e-resources. We acknowledge Metabolome facility (BT/ INF/22/SP28268/2018) at NIPGR for phytochemical analysis.

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Authors and Affiliations

  1. National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi, 110067, India

    Jogindra Naik, Ruchika Rajput & Ashutosh Pandey

  2. Chair of Genetics and Genomics of Plants, Bielefeld University, 33615, Bielefeld, Germany

    Boas Pucker & Ralf Stracke

  3. Evolution and Diversity, Department of Plant Sciences, University of Cambridge, CB2 3EA, Cambridge, UK

    Boas Pucker

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  1. Jogindra Naik
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  2. Ruchika Rajput
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  3. Boas Pucker
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  4. Ralf Stracke
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  5. Ashutosh Pandey
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AP conceived the idea and designed the research. JN, RR and BP conducted experiments. JN and AP interpreted the data. JN, RS and AP wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Ashutosh Pandey.

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The nucleotide sequences reported in this article have been submitted to GenBank under the accession numbers [MW176076 to MW176081].

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Naik, J., Rajput, R., Pucker, B. et al. The R2R3-MYB transcription factor MtMYB134 orchestrates flavonol biosynthesis in Medicago truncatula. Plant Mol Biol 106, 157–172 (2021). https://doi.org/10.1007/s11103-021-01135-x

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