Conserved C3H/APX bifunctionality coordinates lignin deposition and plant growth in Brachypodium and Populus

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ID: 314815
2026
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Abstract
Cytosolic ascorbate peroxidases (APXs) have been proposed to have bifunctional 4-coumarate 3-hydroxylase (C3H) activity, linking redox regulation to lignin biosynthesis in plants. Although this dual role has been shown in vitro, in vivo validation remains limited. Here, we used CRISPR/Cas9 gene editing to knock out cytosolic 4-coumarate 3-hydroxylase/ascorbate peroxidase (C3H/APX) genes in Brachypodium distachyon and poplar (Populus tremula × P. alba). In Brachypodium, BdC3H/APX1 catalyzed the ascorbate-dependent hydroxylation of 4-coumarate to caffeate in vitro. Loss of BdC3H/APX1 function led to reduced lignin content, altered monomer composition, elevated H2O2 levels, and impaired growth, while double monoallelic knockouts of BdC3H/APX1 combined with a biallelic BdC3H/APX2 mutation (Bdc3h/apx1&2) exhibited severe developmental defects. Exogenous caffeate and ferulate rescued the growth and lignin phenotype of the Bdc3h/apx1 knockout mutants, whereas catalase reduced H2O2 without restoring plant growth. Similarly, CRISPR/Cas9-mediated PtC3H/APX1 knockout in poplar resulted in stunted growth and altered lignin composition, while the double Ptc3h/apx1&2 mutants were unable to regenerate from tissue culture. These results provide in vivo evidence of C3H/APX bifunctionality, suggesting that perturbed lignin biosynthesis is the primary cause of the growth defects typically observed in C3H/APX-deficient plants.
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Authors Weiwei Zhu, Harmeet Singh-Bakala, Bo Liu, William Bewg, Max Bentelspacher, Rachel Weber, María Ángeles Peláez‐Vico, M Chen, Chung‐Jui Tsai, Ron Mittler, Bing Yang, Jaime Barros
Journal Plant physiology and biochemistry : PPB
Year 2026
DOI
10.1093/plphys/kiag293
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