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. 2024 Aug 1;43(18):4092–4109. doi: 10.1038/s44318-024-00181-7

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A) Representative UHPLC-UV chromatograms of 2-month-old wild-type, ΔPpCYP73A48, ΔPpCYP73A49, and ΔPpCYP73A48/CYP73A49 crude extracts showing the loss of UV-absorbing molecules in the double mutants. (B) Relative levels of threonate and shikimate phenolic esters in 2-month-old gametophores. pC2T, p-coumaroyl-2-O-threonate; pC4T, p-coumaroyl-4-O-threonate; C2T, caffeoyl-2-O-threonate; C4T, caffeoyl-4-O-threonate; pC5S, p-coumaroyl-5-O-shikimate. (C) Quantification of hydroxycinnamic acids from saponified extracts of 2-month-old gametophores. (D) Compositional analysis of the cuticular polymer of 2-month-old gametophores. (E) Total amount of cuticular monomers from gametophores. (F) Representative pictures of 2-month-old wild-type, ΔPpCYP73A48 and ΔPpCYP73A49 gametophores after toluidine blue permeability assay. (G) Quantification of permeability to toluidine blue in 2-month-old gametophores. Results in panels (B), (C), (D), (E), and (F) are the mean ± SEM of three independent WT biological replicates and three independent mutant lines. WT versus mutant unpaired t test adjusted P-value: *P < 0.05; **P < 0.01; ***P < 0.001. nd, not detected. Source data are available online for this figure.