8587e6af6ce2a8875acca663bd4f832c 26TV08_PlantPhysiol_Riechmann_pub.pdf e232c73025e88b6da58c6cbf2813ab8cf0656b81 26TV08_PlantPhysiol_Riechmann_pub.pdf f1733e5515f21f4f64ca9094819cc57f4228d6f7d7bb3aed0a762d4e55b56690 26TV08_PlantPhysiol_Riechmann_pub.pdf Title: The non-conventional peptidome of Arabidopsis flower development Subject: DOI: 10.1093/plphys/kiag047; Plant Physiology, 200, 2, 2026-02-20.; Abstract: Unraveling the complexities of genomic data has revealed that the protein coding capacity of eukaryotic genomes has been underestimated. Eukaryotic genomes contain numerous unannotated short open reading frames (sORFs) that, localized in different types of RNA molecules, including long non-coding RNAs, may encode and produce biologically functional peptides. This study focuses on the characterization of the Arabidopsis (Arabidopsis thaliana) sORF-derived flower peptidome, using the floral homeotic mutants apetala1, apetala2, apetala3, pistillata, and agamous in comparison to the wild type. For peptide identification by mass spectrometry (MS), we created an extensive database of hypothetical Arabidopsis peptides, which comprised putative sORF-encoded peptides from intergenic regions, untranslated regions, “non-coding” RNAs and other transcripts. In total, 1,874 hypothetical peptides were detected by MS, of which 132 were selected as higher-confidence peptides for further studies. Sixty of these higher-confidence peptides were predicted to be specifically expressed, or at least enriched, in particular floral organs. Approximately 25% of them belonged to putative gene families in A. thaliana, and 103 had potential homologs in other plant species. Additionally, distinct gene expression patterns were observed, in many cases consisting of specific expression in stamens during flower development. Keywords: Author: Raquel Álvarez-Urdiola Creator: Servigistics Arbortext Advanced Print Publisher 11.1.4546/W-x64 Producer: PDFlib+PDI 9.0.7p3 (C++/Win64); modified using iTextSharp.LGPLv2.Core 3.7.4.0 CreationDate: Fri Feb 20 08:53:33 2026 CET ModDate: Fri Feb 20 22:31:57 2026 CET Custom Metadata: yes Metadata Stream: yes Tagged: yes UserProperties: no Suspects: no Form: none JavaScript: no Pages: 20 Encrypted: no Page size: 595.276 x 782.362 pts Page rot: 0 File size: 2091186 bytes Optimized: no PDF version: 1.6 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- OFGPVT+NotoSans-Italic CID TrueType Identity-H yes yes yes 50 0 BFIJBM+NotoSans-Regular CID TrueType Identity-H yes yes yes 46 0 IYNXPR+NotoSans-Bold CID TrueType Identity-H yes yes yes 130 0 RBWJHL+Oswald-SemiBold CID TrueType Identity-H yes yes yes 2404 0 QTXEMC+Oswald-Light CID TrueType Identity-H yes yes yes 2405 0 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acid sequence features Destination: seckiag047-s2.3 Item: Title: Several SEPs belong to putative peptide families in Arabidopsis Destination: seckiag047-s2.4 Item: Title: Amino acid sequences of SEPs are conserved across species Destination: seckiag047-s2.5 Item: Title: SEPs identified in floral buds show differential gene expression across tissues Destination: seckiag047-s2.6 Item: Title: Expression patterns for selected higher-confidence peptides determined by reporter gene fusions Destination: seckiag047-s2.7 Item: Title: Discussion Destination: seckiag047-s3 Item: Title: Materials and methods Destination: seckiag047-s4 Children: Item: Title: Plant lines, growth conditions, and tissue collection Destination: seckiag047-s4.1 Item: Title: Peptide extraction Destination: seckiag047-s4.2 Children: Item: Title: Ultrafiltration Destination: seckiag047-s4.2.1 Item: Title: Reverse phase chromatography Destination: seckiag047-s4.2.2 Item: Title: Liquid chromatography tandem MS (LC-MS/MS) Destination: seckiag047-s4.3 Children: Item: Title: Sample preparation Destination: seckiag047-s4.3.1 Item: Title: Chromatography and MS analysis Destination: seckiag047-s4.3.2 Item: Title: Data analysis Destination: seckiag047-s4.3.3 Item: Title: Reference database Destination: seckiag047-s4.3.4 Item: Title: Re-annotation of TIS and identification of amino acid sequence features Destination: seckiag047-s4.4 Item: Title: Sequence conservation analyses Destination: seckiag047-s4.5 Children: Item: Title: Analysis of related sequences within the sORF datasets Destination: seckiag047-s4.5.1 Item: Title: Homology analysis Destination: seckiag047-s4.5.2 Item: Title: Gene expression of SEPs in different tissues Destination: seckiag047-s4.6 Children: Item: Title: RNA extraction and cDNA obtention Destination: seckiag047-s4.6.1 Item: Title: qPCR primer design and testing Destination: seckiag047-s4.6.2 Item: Title: Quantitative real time PCR BioMark System Destination: seckiag047-s4.6.3 Item: Title: Generation of reporter lines Destination: seckiag047-s4.7 Children: Item: Title: Gene selection Destination: seckiag047-s4.7.1 Item: Title: Promoter amplification Destination: seckiag047-s4.7.2 Item: Title: Gateway vectors Destination: seckiag047-s4.7.3 Item: Title: Bacterial strains Destination: seckiag047-s4.7.4 Item: Title: Plant transformation Destination: seckiag047-s4.7.5 Item: Title: GUS staining Destination: seckiag047-s4.8 Item: Title: Accession numbers Destination: seckiag047-s4.9 Item: Title: Acknowledgments Destination: acknow Item: Title: Author contributions Destination: seckiag047-s5 Item: Title: Supplementary material Destination: seckiag047-s6 Item: Title: Funding Destination: seckiag047-s7 Item: Title: Conflicts of interest Destination: seckiag047-s8 Item: Title: Data availability Destination: seckiag047-s9 Item: Title: References Destination: reflist Info: Title: The non-conventional peptidome of Arabidopsis flower development Author: Raquel Álvarez-Urdiola Subject: DOI: 10.1093/plphys/kiag047; Plant Physiology, 200, 2, 2026-02-20.; Abstract: Unraveling the complexities of genomic data has revealed that the protein coding capacity of eukaryotic genomes has been underestimated. Eukaryotic genomes contain numerous unannotated short open reading frames (sORFs) that, localized in different types of RNA molecules, including long non-coding RNAs, may encode and produce biologically functional peptides. This study focuses on the characterization of the Arabidopsis (Arabidopsis thaliana) sORF-derived flower peptidome, using the floral homeotic mutants apetala1, apetala2, apetala3, pistillata, and agamous in comparison to the wild type. For peptide identification by mass spectrometry (MS), we created an extensive database of hypothetical Arabidopsis peptides, which comprised putative sORF-encoded peptides from intergenic regions, untranslated regions, “non-coding” RNAs and other transcripts. In total, 1,874 hypothetical peptides were detected by MS, of which 132 were selected as higher-confidence peptides for further studies. Sixty of these higher-confidence peptides were predicted to be specifically expressed, or at least enriched, in particular floral organs. Approximately 25% of them belonged to putative gene families in A. thaliana, and 103 had potential homologs in other plant species. Additionally, distinct gene expression patterns were observed, in many cases consisting of specific expression in stamens during flower development. 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University Press The non-conventional peptidome of Arabidopsis flower development Raquel Álvarez-Urdiola José Tomás Matus Víctor M. González-Miguel Martí Bernardo-Faura José Luis Riechmann DOI: 10.1093/plphys/kiag047; Plant Physiology, 200, 2, 2026-02-20.; Abstract: Unraveling the complexities of genomic data has revealed that the protein coding capacity of eukaryotic genomes has been underestimated. Eukaryotic genomes contain numerous unannotated short open reading frames (sORFs) that, localized in different types of RNA molecules, including long non-coding RNAs, may encode and produce biologically functional peptides. This study focuses on the characterization of the Arabidopsis (Arabidopsis thaliana) sORF-derived flower peptidome, using the floral homeotic mutants apetala1, apetala2, apetala3, pistillata, and agamous in comparison to the wild type. For peptide identification by mass spectrometry (MS), we created an extensive database of hypothetical Arabidopsis peptides, which comprised putative sORF-encoded peptides from intergenic regions, untranslated regions, “non-coding” RNAs and other transcripts. In total, 1,874 hypothetical peptides were detected by MS, of which 132 were selected as higher-confidence peptides for further studies. Sixty of these higher-confidence peptides were predicted to be specifically expressed, or at least enriched, in particular floral organs. Approximately 25% of them belonged to putative gene families in A. thaliana, and 103 had potential homologs in other plant species. Additionally, distinct gene expression patterns were observed, in many cases consisting of specific expression in stamens during flower development. © The Author(s) 2026. Published by Oxford University Press on behalf of American Society of Plant Biologists. Journal Plant Physiology © The Author(s) 2026. 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