# Genome-wide analysis of the R2R3-MYB family reveals potential regulators of lignin and tricin metabolism in the model grass Setaria viridis

**Authors:** Giovanni Victorio Cerruti, Lucas Xavier da Cunha, Alice Cristine Cursino, Marcella Siqueira Simões, Igor Cesarino

PMC · DOI: 10.1007/s00438-026-02355-w · Molecular Genetics and Genomics · 2026-02-07

## TL;DR

This study identifies R2R3-MYB transcription factors in Setaria viridis that may regulate lignin and tricin metabolism, offering insights into grass-specific lignification.

## Contribution

The paper provides the first genome-wide analysis of R2R3-MYB regulators in Setaria viridis, identifying candidates for lignin and tricin metabolism.

## Key findings

- 132 R2R3-MYB genes were identified in Setaria viridis, grouped into 43 subgroups.
- Four candidate SvMYBs showed expression patterns linked to lignin biosynthesis and were highly expressed in tissues undergoing lignification.
- Three SvMYBs activated promoters of lignin and tricin biosynthetic genes in transactivation assays.

## Abstract

R2R3-MYBs constitute one of the largest families of plant transcription factors and several members are involved in the regulation of distinct branches of the phenylpropanoid pathway, acting as activators or repressors of the biosynthesis of a plethora of specialized metabolites. Although R2R3-MYBs have been extensively characterized as regulators of lignin deposition in distinct cellular contexts and species, still little is known about their roles in regulating specific aspects of grass lignification. Here, we report on the genome-wide characterization of the R2R3-MYB family in the model grass Setaria viridis and identification of members potentially involved in the regulation of lignin/tricin metabolism. A total of 132 genes encoding R2R3-MYBs were found in S. viridis, which clustered in 43 well-supported subgroups. Comprehensive in silico expression, co-expression, and RT-qPCR analyses allowed the identification of 4 candidate SvMYBs that showed (i) similar expression profiles to that observed for lignin biosynthetic genes in a set of different organs/conditions of S. viridis; (ii) similar expression patterns to that of lignin biosynthetic genes along the S. viridis elongating internode; (iii) co-expression with several phenylpropanoid- and lignin-related genes in public transcriptomic databases; (iv) high expression levels in the top of the S. viridis elongating internode, a tissue undergoing active lignification. Three of these SvMYBs activated the promoters of lignin and tricin biosynthetic genes in transactivation assays using tobacco protoplasts. Altogether, our results suggest that these three transcription factors control grass-specific aspects of lignin deposition and further studies might confirm their ability to control lignin deposition and tricin metabolism in S. viridis.

The online version contains supplementary material available at 10.1007/s00438-026-02355-w.

## Linked entities

- **Chemicals:** lignin (PubChem CID 175586), tricin (PubChem CID 5281702)
- **Species:** Setaria viridis (taxon 4556), Nicotiana tabacum (taxon 4097)

## Full-text entities

- **Genes:** 4CL1 (4-coumarate:CoA ligase 1) [NCBI Gene 841593] {aka 4-COUMARATE:COA LIGASE, 4-COUMARATE:COA LIGASE 1, 4-coumarate:CoA ligase 1, 4CL.1, ARABIDOPSIS THALIANA 4-COUMARATE:COA LIGASE 1, AT4CL1}, MYB42 (myb domain protein 42) [NCBI Gene 826844] {aka AtMYB42, T4C9.190, T4C9_190, myb domain protein 42}, MYB49 (myb domain protein 49) [NCBI Gene 835511] {aka AtMYB49, MDK4.5, MDK4_5, myb domain protein 49}, MYB99 (myb domain protein 99) [NCBI Gene 836353] {aka ATMYB99, ATMYBCU15, MMI9.18, MMI9_18, myb domain protein 99}, MYB61 (myb domain protein 61) [NCBI Gene 837480] {aka ARABIDOPSIS THALIANA MYB DOMAIN PROTEIN 61, ATMYB61, F14J9.20, F14J9_20, myb domain protein 61}, PAP1 (phosphatidic acid phosphatase 1) [NCBI Gene 814646] {aka ATLPP1, ATPAP1, F10A8.6, F10A8_6, LIPID PHOSPHATE PHOSPHATASE 1, LPP1}, MYB57 (myb domain protein 57) [NCBI Gene 821113] {aka ATMYB57, F4P13.8, F4P13_8, myb domain protein 57}, MYB88 (myb domain protein 88) [NCBI Gene 814812] {aka AtMYB88, T20F6.4, T20F6_4, myb domain protein 88}, MYB40 (myb domain protein 40) [NCBI Gene 831284] {aka AtMYB40, F18O22.130, F18O22_130, myb domain protein 40}, MYB85 (myb domain protein 85) [NCBI Gene 828364] {aka AtMYB85, T12H17.70, T12H17_70, myb domain protein 85}, PAP1 (production of anthocyanin pigment 1) [NCBI Gene 842120] {aka ATMYB75, F25P12.92, F25P12_92, MYB DOMAIN PROTEIN 75, MYB75, MYELOBLASTOSIS PROTEIN 75}, TDF1 (Duplicated homeodomain-like superfamily protein) [NCBI Gene 822477] {aka ATMYB35, DEFECTIVE IN MERISTEM DEVELOPMENT AND FUNCTION 1, MYB DOMAIN PROTEIN 35, MYB35}, MYB26 (myb domain protein 26) [NCBI Gene 820602] {aka ATMYB26, MALE STERILE 35, MS35, myb domain protein 26}, MYB86 (myb domain protein 86) [NCBI Gene 832723] {aka ATMYB4, ATMYB4-old, ATMYB86, myb domain protein 4, myb domain protein 86}, MYB43 (myb domain protein 43) [NCBI Gene 831522] {aka AtMYB43, MTG13.12, MTG13_12, myb domain protein 43}, MYB10 (myb domain protein 10) [NCBI Gene 820464] {aka AtMYB10, myb domain protein 10}, MYB5 (myb domain protein 5) [NCBI Gene 820556] {aka ABC transporter of the mitochondrion 2, ATM2, ATMYB5, myb domain protein 5}, TT4 (Chalcone and stilbene synthase family protein) [NCBI Gene 831241] {aka ATCHS, CHALCONE SYNTHASE, CHALCONE/STILBENE SYNTHASE, CHS, MAC12.28, MAC12_28}, AtMYB103 (myb domain protein 103) [NCBI Gene 842694] {aka MYB103, T12P18.7, T12P18_7, myb domain protein 103}, MYB20 (myb domain protein 20) [NCBI Gene 842938] {aka AtMYB20, T6J19.5, T6J19_5, myb domain protein 20}, MYB89 (myb domain protein 89) [NCBI Gene 833966] {aka AtMYB89, MIJ24.170, MIJ24_170, myb domain protein 89}, MYB46 (myb domain protein 46) [NCBI Gene 831127] {aka ATMYB46, MYB DOMAIN PROTEIN 46, T24H18.40, T24H18_40, myb domain protein 46}, MYB68 (myb domain protein 68) [NCBI Gene 836708] {aka ATMYB68, MPA24.14, MPA24_14, MYB DOMAIN PROTEIN 68, myb domain protein 68}, MYB37 (myb domain protein 37) [NCBI Gene 832364] {aka ATMYB37, RAX1, REGULATOR OF AXILLARY MERISTEMS 1, T20O7.2, T20O7_2, myb domain protein 37}, MYB58 (myb domain protein 58) [NCBI Gene 838219] {aka ATMYB58, F3O9.29, F3O9_29, MYB DOMAIN PROTEIN 58, myb domain protein 58}, MYB82 (myb domain protein 82) [NCBI Gene 835337] {aka AtMYB82, myb domain protein 82}, MYB6 (myb domain protein 6) [NCBI Gene 826530] {aka ATMYB6, AtMYB6, T15G18.120, T15G18_120, myb domain protein 6}, MYB36 (myb domain protein 36) [NCBI Gene 835866] {aka AtMYB36, MUA2.20, MUA2_20, myb domain protein 36}, MYB12 (myb domain protein 12) [NCBI Gene 819359] {aka ATMYB12, MYB DOMAIN PROTEIN 12, PFG1, PRODUCTION OF FLAVONOL GLYCOSIDES 1, T30B22.24, myb domain protein 12}, MYB47 (myb domain protein 47) [NCBI Gene 838453] {aka AtMYB47, F6A14.18, F6A14_18, myb domain protein 47}
- **Diseases:** developmental defects (MESH:D000094602)
- **Chemicals:** stilbene (MESH:D013267), phenolic acid (MESH:C017616), proanthocyanidin (MESH:C013221), chlorogenic acid (MESH:D002726), sinapyl alcohol (MESH:C496130), flavonoid (MESH:D005419), chrysoeriol (MESH:C007054), flavonol (MESH:C041477), phenylalanine (MESH:D010649), nitrogen (MESH:D009584), glucosinolate (MESH:D005961), anthocyanin (MESH:D000872), catechin (MESH:D002392), sugar (MESH:D000073893), apigenin (MESH:D047310), polyphenols (MESH:D059808), tricin (MESH:C017769), RQ1 (-), S-lignin (MESH:D008031), S (MESH:D013455), flavonols (MESH:D044948), luteolin (MESH:D047311)
- **Species:** Brachypodium distachyon (annual false brome, species) [taxon 15368], Limosilactobacillus fermentum (species) [taxon 1613], Nicotiana tabacum (American tobacco, species) [taxon 4097], Eucalyptus grandis (rose gum, species) [taxon 71139], Homo sapiens (human, species) [taxon 9606], Sorghum bicolor (broomcorn, species) [taxon 4558], Populus trichocarpa (black cottonwood, species) [taxon 3694], Vitis vinifera (wine grape, species) [taxon 29760], Arabidopsis thaliana (mouse-ear cress, species) [taxon 3702], Oryza sativa (Asian cultivated rice, species) [taxon 4530], Setaria viridis (species) [taxon 4556], Panicum virgatum (switchgrass, species) [taxon 38727], Setaria italica (foxtail millet, species) [taxon 4555], Cucumis sativus (cucumber, species) [taxon 3659], Setaria viridis subsp. viridis (subspecies) [taxon 1043315], Solanum lycopersicum (tomato, species) [taxon 4081]
- **Cell lines:** -2 — Homo sapiens (Human), Colon carcinoma, Cancer cell line (CVCL_A628)

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Source: https://tomesphere.com/paper/PMC12882868