c21f80dc4d19f4fc4f4949ac9c86e485 JouExp_a2025v76n15p4340.pdf 203d524a445d303aa7f6a86126b80d5271da642e JouExp_a2025v76n15p4340.pdf 2ea3aa06d0885dd0809eb7c8a28813e60ab399d07ef00d07700dcf2dd1b0fd3b JouExp_a2025v76n15p4340.pdf Title: The CAPE1 peptide confers resistance against bacterial wilt in tomato Subject: DOI: 10.1093/jxb/eraf145 Journal of Experimental Botany, Volume. XX, Issue. XX, 3 April 2025 Abstract Bacterial wilt caused by Ralstonia solanacearum is one of the most destructive bacterial diseases for which no effective treatment exists. There is an urgent need to understand the basis of resistance against this pathogen in order to engineer efficient strategies in the field. We previously demonstrated that resistant tomato plants limit bacterial movement in the apoplast and the xylem. As a first step to dissect the underlying mechanisms, we analysed the apoplast proteome upon challenge with R. solanacearum in the susceptible tomato cultivar Marmande and the resistant cultivar Hawaii 7996. Here, we described the xylem proteome in these same cultivars and compared it with the apoplastic proteome, revealing variety-dependent and infection-dependent changes. This proteomic analysis led to the identification of pathogenesis-related 1 (PR1) proteins as highly induced upon infection. Since PR1b was the most abundant PR1 protein in both the apoplast and the xylem, we concentrated on this family member to study the role of PR1s in the interaction between tomato and R. solanacearum. Surprisingly, lack of PR1b resulted in enhanced resistance to R. solanacearum in tomato, which could be due to an up-regulation of homologous genes in a compensatory effect as has been reported before. PR1 processing by an unknown protease in tomato results in the generation of the CAPE peptide. Treatment of tomato plants with the CAPE1 peptide resulted in restriction of R. solanacearum growth, via defence gene reprogramming. Future work in the lab will help determine which tomato secreted proteases cleave PR1s to generate CAPEs. Author: Weiqi Zhang,Marc Planas-Marquès,Moyan Liang,Qingshan Zhang,Annemarie Vermeulen,Farnusch Kaschani,Markus Kaiser,Frank L. W. Takken,Nuria S. Coll,Marc Valls Creator: Adobe InDesign 15.1 (Windows) Producer: Adobe PDF Library 15.0; modified using iTextSharp.LGPLv2.Core 3.7.4.0 CreationDate: Sat May 10 11:26:47 2025 CEST ModDate: Wed Jul 15 17:46:17 2026 CEST Custom Metadata: no Metadata Stream: yes Tagged: yes UserProperties: no Suspects: no Form: AcroForm JavaScript: no Pages: 19 Encrypted: no Page size: 612.283 x 790.866 pts Page rot: 0 File size: 6567072 bytes Optimized: no PDF version: 1.6 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- HLHWMP+HelveticaNeueLTStd-Lt Type 1C Custom yes yes yes 2100 0 WGYAHF+Helvetica-LightOblique Type 1C Custom yes yes no 2101 0 NHWDKX+HelveticaNeueLTStd-LtIt Type 1C WinAnsi yes yes yes 2102 0 NHWDKX+HelveticaNeueLTStd-LtIt Type 1C Custom yes yes yes 65 0 HLHWMP+HelveticaNeueLTStd-Lt Type 1C Custom yes yes yes 55 0 MIJLMA+HelveticaNeueLTStd-Md Type 1C WinAnsi yes yes yes 2103 0 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and valid SignatureMatches: PDF-hul MIMEtype: application/pdf Profile: Tagged PDF PDFMetadata: Objects: 2157 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: false CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Language: en-US Outlines: Item: Title: The CAPE1 peptide confers resistance against bacterial wilt in tomato Children: Item: Title: Introduction Item: Title: Materials and methods Children: Item: Title: Plant material and growth conditions Item: Title: Bacterial material, plant inoculations, and pathogenicity assays Item: Title: Xylem sap collection, pooling, and concentration Item: Title: Proteomic analyses Children: Item: Title: Sample clean-up for liquid chromatography–mass spectrometry Item: Title: Liquid chromatography–tandem mass spectrometry Item: Title: Peptide and protein identification Item: Title: Sequence comparison and phylogenetic analyses of PR1 proteins Item: Title: Cloning of the PR1 proteins Item: Title: Transient expression PR1 proteins in Nicotiana benthamiana Item: Title: Generation of stable PR1b overexpression tomato plants Item: Title: Purification of PR1b proteins and derivatives Item: Title: Generation of CRISPR/Cas9 pr1b mutant tomato plants Item: Title: Extraction of apoplast and total extract from tomato leaves and N. benthamiana leaves Item: Title: Confocal microscopy Item: Title: Treatment plants with CAPE1 peptide Item: Title: Antimicrobial assay in vitro Item: Title: RNA extraction and quantitative reverse transcription–PCR analysis Item: Title: Statistical analyses Item: Title: Results Children: Item: Title: Xylem sap and apoplast proteomes reveal high accumulation of PR1 proteins upon R. solanacearum infection Item: Title: Characterization of the tomato PR1 family and its conservation Item: Title: PR1b involvement in tomato defence Item: Title: Role of the CAPE1 peptide in response to R. solanacearum Item: Title: Discussion Item: Title: Supplementary data Item: Title: Acknowledgements Item: Title: References Info: Title: The CAPE1 peptide confers resistance against bacterial wilt in tomato Author: Weiqi Zhang,Marc Planas-Marquès,Moyan Liang,Qingshan Zhang,Annemarie Vermeulen,Farnusch Kaschani,Markus Kaiser,Frank L. W. Takken,Nuria S. Coll,Marc Valls Subject: DOI: 10.1093/jxb/eraf145 Journal of Experimental Botany, Volume. XX, Issue. XX, 3 April 2025 Abstract Bacterial wilt caused by Ralstonia solanacearum is one of the most destructive bacterial diseases for which no effective treatment exists. There is an urgent need to understand the basis of resistance against this pathogen in order to engineer efficient strategies in the field. We previously demonstrated that resistant tomato plants limit bacterial movement in the apoplast and the xylem. As a first step to dissect the underlying mechanisms, we analysed the apoplast proteome upon challenge with R. solanacearum in the susceptible tomato cultivar Marmande and the resistant cultivar Hawaii 7996. Here, we described the xylem proteome in these same cultivars and compared it with the apoplastic proteome, revealing variety-dependent and infection-dependent changes. This proteomic analysis led to the identification of pathogenesis-related 1 (PR1) proteins as highly induced upon infection. Since PR1b was the most abundant PR1 protein in both the apoplast and the xylem, we concentrated on this family member to study the role of PR1s in the interaction between tomato and R. solanacearum. Surprisingly, lack of PR1b resulted in enhanced resistance to R. solanacearum in tomato, which could be due to an up-regulation of homologous genes in a compensatory effect as has been reported before. PR1 processing by an unknown protease in tomato results in the generation of the CAPE peptide. Treatment of tomato plants with the CAPE1 peptide resulted in restriction of R. solanacearum growth, via defence gene reprogramming. Future work in the lab will help determine which tomato secreted proteases cleave PR1s to generate CAPEs. 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Adobe Registry: Identity Supplement: 0 FontDescriptor: FontName: WUMKWG+Arimo-Bold Flags: Symbolic FontBBox: -482, -376, 1304, 1033 FontFile2: true XMP: application/pdf The CAPE1 peptide confers resistance against bacterial wilt in tomato DOI: 10.1093/jxb/eraf145 Journal of Experimental Botany, Volume. XX, Issue. XX, 3 April 2025 Abstract Bacterial wilt caused by Ralstonia solanacearum is one of the most destructive bacterial diseases for which no effective treatment exists. There is an urgent need to understand the basis of resistance against this pathogen in order to engineer efficient strategies in the field. We previously demonstrated that resistant tomato plants limit bacterial movement in the apoplast and the xylem. As a first step to dissect the underlying mechanisms, we analysed the apoplast proteome upon challenge with R. solanacearum in the susceptible tomato cultivar Marmande and the resistant cultivar Hawaii 7996. Here, we described the xylem proteome in these same cultivars and compared it with the apoplastic proteome, revealing variety-dependent and infection-dependent changes. This proteomic analysis led to the identification of pathogenesis-related 1 (PR1) proteins as highly induced upon infection. Since PR1b was the most abundant PR1 protein in both the apoplast and the xylem, we concentrated on this family member to study the role of PR1s in the interaction between tomato and R. solanacearum. Surprisingly, lack of PR1b resulted in enhanced resistance to R. solanacearum in tomato, which could be due to an up-regulation of homologous genes in a compensatory effect as has been reported before. PR1 processing by an unknown protease in tomato results in the generation of the CAPE peptide. Treatment of tomato plants with the CAPE1 peptide resulted in restriction of R. solanacearum growth, via defence gene reprogramming. Future work in the lab will help determine which tomato secreted proteases cleave PR1s to generate CAPEs. © The Author(s) 2025. Published by Oxford University Press on behalf of the Society for Experimental Biology. Weiqi Zhang,Marc Planas-Marquès,Moyan Liang,Qingshan Zhang,Annemarie Vermeulen,Farnusch Kaschani,Markus Kaiser,Frank L. W. Takken,Nuria S. Coll,Marc Valls  Bacterial wilt, PR1 proteins, Ralstonia solanacearum, Solanum lycopersicum, tomato disease resistance, xylem proteome 2025-05-10T14:56:47+05:30 Adobe InDesign 15.1 (Windows) 2026-07-15T15:46:17+00:00 2026-07-15T15:46:17+00:00 uuid:8a67f32e-3888-44da-abf7-8839b9eb4003 xmp.did:840A93442E2068118083C1556DCA13C0 xmp.id:18e45c2f-bd2d-ff4b-8a80-e20bb4b01d73 proof:pdf 1 xmp.iid:083f03c7-ca15-1c41-b885-af5bef3feaca xmp.did:0b6d3d89-3aae-be43-86eb-8b1478d77bde default converted from application/x-indesign to application/pdf Adobe InDesign 15.1 (Windows) 2025-05-10T14:56:47+05:30 converted uuid:f7b01c6e-2f07-4162-9e11-00c2b68c9f72 converted to PDF/A-3a Preflight 2025-05-10T14:58:24+05:30 True Adobe PDF Library 15.0; modified using iTextSharp.LGPLv2.Core 3.7.4.0 False http://ns.adobe.com/xap/1.0/mm/ xmpMM XMP Media Management Schema internal UUID based identifier for specific incarnation of a document InstanceID URI internal The common identifier for all versions and renditions of a document. 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Annotation: Subtype: Link Rect: 44, 90, 236, 81 Flags: Print AppearanceDictionary: true Destination: 43 Annotation: Subtype: Link Rect: 313, 693, 443, 684 Flags: Print AppearanceDictionary: true Destination: 67 Annotation: Subtype: Link Rect: 313, 663, 571, 654 Flags: Print AppearanceDictionary: true Destination: 70 Annotation: Subtype: Link Rect: 313, 654, 530, 645 Flags: Print AppearanceDictionary: true Destination: 70 Annotation: Subtype: Link Rect: 313, 624, 571, 615 Flags: Print AppearanceDictionary: true Destination: 186 Annotation: Subtype: Link Rect: 313, 615, 492, 606 Flags: Print AppearanceDictionary: true Destination: 186 Annotation: Subtype: Link Rect: 313, 585, 478, 576 Flags: Print AppearanceDictionary: true Destination: 189 Annotation: Subtype: Link Rect: 313, 555, 570, 546 Flags: Print AppearanceDictionary: true Destination: 43 Annotation: Subtype: Link Rect: 313, 516, 571, 507 Flags: Print AppearanceDictionary: true Destination: 43 Annotation: Subtype: Link Rect: 313, 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