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Although it remains one of the most characterized C4–CAM species, many aspects of this dual system are still unresolved. This expert view summarizes recent advances in understanding C4–CAM integration in Portulaca, highlighting key genomic, transcriptomic, biochemical, and physiological insights, and outlines the most pressing questions about the functioning and evolution of this combined CCM. Future directions for explaining how these convergent pathways emerged in the same lineage include functional studies on key regulatory elements, spatially resolved analyses for protein distribution and metabolic fluxes across leaves, an updated phylogeny of the family, and the integration of molecular and physiological data. So far, it is evident that expressing a weak CAM cycle in a C4 crop might be possible and could entail increased survival, possibly contributing to higher productivity in a future with climate change. Keywords: Abiotic stress; carbon-concentrating mechanism; C4 photosynthesis; Crassulacean acid metabolism; drought stress; facultative CAM; PEPC; phosphoenolpyruvate carboxylase; photosynthesis; Portulaca oleracea Author: Renata Callegari Ferrari 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: Sat Jul 11 12:40:34 2026 CEST ModDate: Thu Oct 22 12:34:59 2026 CEST Custom Metadata: yes Metadata Stream: yes Tagged: yes UserProperties: no Suspects: no Form: AcroForm JavaScript: no Pages: 12 Encrypted: no Page size: 612.283 x 790.866 pts Page rot: 0 File size: 1353474 bytes Optimized: no PDF version: 1.6 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- YQMQXS+MicrosoftSansSerif CID TrueType Identity-H yes yes yes 34 0 JJTJHX+HelveticaNeueLTStd-LtIt Type 1C Custom yes yes yes 39 0 RARYCV+HelveticaNeueLTStd-Lt Type 1C Custom yes yes yes 38 0 QCCRGN+HelveticaNeueLTStd-Md Type 1C Custom yes yes yes 1039 0 AVQYMM+HelveticaNeueLTStd-Bd Type 1C Custom yes yes yes 37 0 NPJOFU+HelveticaNeueLTStd-BdIt Type 1C Custom yes yes yes 60 0 CRTMJA+BemboMTStd-Regular Type 1C Custom yes yes yes 35 0 JCMDBA+STIXGeneral-Regular Type 1C Custom yes yes yes 43 0 ZTFMEU+BemboMTStd-Italic Type 1C Custom yes yes yes 36 0 UCBNYB+ArialMT TrueType WinAnsi yes yes no 1 0 VEAKYP+TimesNewRomanPSMT CID TrueType Identity-H yes yes yes 33 0 LucidaMath-Symbol Type 1 Builtin yes no yes 44 0 DBZOFD+TimesNewRomanPS-ItalicMT CID TrueType Identity-H yes yes yes 57 0 NUTRXZ+HelveticaNeueLTStd-Roman Type 1C Custom yes yes yes 58 0 OIODNP+HelveticaNeueLTStd-It Type 1C Custom yes yes yes 59 0 CFHSWG+SourceSansPro-Regular Type 1C Custom yes yes yes 77 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2026-10-23 04:10:47 CEST RepresentationInformation: jouexpbot_a2026m7v77n14p4316.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2026-10-22 12:38:42 CEST Size: 1353474 Format: PDF Version: 1.6 Status: Well-Formed, but not valid SignatureMatches: PDF-hul ErrorMessage: Invalid indirect destination - referenced object 'corerag161-FM2' cannot be found ID: PDF-HUL-149 ErrorMessage: Invalid indirect destination - referenced object 'tblerag161-box1' cannot be found ID: PDF-HUL-149 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'tblerag161-box1' cannot be found ID: PDF-HUL-122 Offset: 1280341 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'corerag161-FM2' cannot be found ID: PDF-HUL-122 Offset: 1280341 MIMEtype: application/pdf Profile: Tagged PDF PDFMetadata: Objects: 1285 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: false CenterWindow: false DisplayDocTitle: true NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: OneColumn PageMode: UseOutlines Language: en Outlines: Item: Title: Open questions on facultative C4 CAM photosynthesis in Portulaca Destination: head1 Children: Item: Title: Introduction Destination: secerag161-s1 Item: Title: To be or not to be: why plants perform C4 or CAM Destination: secerag161-s2 Item: Title: The best of both worlds: the C4 CAM system Destination: secerag161-s3 Item: Title: How do C4 and CAM pathways co-exist within the same leaf at the biochemical and regulatory levels? Destination: secerag161-s4 Children: Item: Title: What role do regulatory enzymes such as phosphoenolpyruvate carboxylase kinase (PPCK) play in this pathway regulation? Destination: secerag161-s4.1 Item: Title: What transcriptional and post-transcriptional mechanisms regulate the switch between pathways? Destination: secerag161-s4.2 Item: Title: How are metabolic fluxes coordinated? Destination: secerag161-s4.3 Item: Title: What evolutionary processes enabled the convergence of two CCMs within the same genus? Destination: secerag161-s4.4 Item: Title: What selective pressures favoured this convergence? Destination: secerag161-s4.5 Item: Title: What genomic and regulatory elements enabled the integration of these pathways? Destination: secerag161-s4.6 Item: Title: Can this knowledge inform efforts to engineer metabolic flexibility in crops? Destination: secerag161-s5 Children: Item: Title: How might synthetic biology approaches exploit this system for enhanced stress resilience? Destination: secerag161-s5.1 Item: Title: Can aspects of this dual pathway be introduced into other species? Destination: secerag161-s5.2 Item: Title: Conclusion Destination: secerag161-s6 Item: Title: Author contribution Destination: secerag161-s7 Item: Title: Conflict of interest Destination: secerag161-s8 Item: Title: Funding Destination: secerag161-s9 Item: Title: References Destination: reflist Info: Title: Open questions on facultative C4 CAM photosynthesis in Portulaca Author: Renata Callegari Ferrari Subject: DOI: 10.1093/jxb/erag161; Journal of Experimental Botany, 77, 14, 2026-04-10.; Abstract: Plants have evolved carbon-concentrating mechanisms (CCMs) to optimize carbon fixation under environmental conditions that increase photorespiratory and transpiration rates. C4 and Crassulacean acid metabolism (CAM) photosynthesis are the two most widespread CCM examples across land plants and were typically considered to be incompatible due to biochemical, anatomical, evolutionary, and regulatory characteristics. However, Portulaca oleracea (purslane) was the first reported case to challenge this assumption >40 years ago. Although it remains one of the most characterized C4 CAM species, many aspects of this dual system are still unresolved. This expert view summarizes recent advances in understanding C4 CAM integration in Portulaca, highlighting key genomic, transcriptomic, biochemical, and physiological insights, and outlines the most pressing questions about the functioning and evolution of this combined CCM. Future directions for explaining how these convergent pathways emerged in the same lineage include functional studies on key regulatory elements, spatially resolved analyses for protein distribution and metabolic fluxes across leaves, an updated phylogeny of the family, and the integration of molecular and physiological data. So far, it is evident that expressing a weak CAM cycle in a C4 crop might be possible and could entail increased survival, possibly contributing to higher productivity in a future with climate change. Keywords: Abiotic stress; carbon-concentrating mechanism; C4 photosynthesis; Crassulacean acid metabolism; drought stress; facultative CAM; PEPC; phosphoenolpyruvate carboxylase; photosynthesis; Portulaca oleracea 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: Sat Jul 11 13:10:34 CEST 2026 ModDate: Thu Oct 22 12:34:59 CEST 2026 ID: 0x2b00168d5318bf85d0619c0b8804e909, 0x635d1e935f4d60e1f147870e6f3c58d4 Filters: FilterPipeline: FlateDecode FilterPipeline: DCTDecode Images: Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 1456 ImageHeight: 966 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Fonts: Type0: Font: BaseFont: VEAKYP+TimesNewRomanPSMT Encoding: Identity-H ToUnicode: true Font: BaseFont: YQMQXS+MicrosoftSansSerif Encoding: Identity-H ToUnicode: true Font: BaseFont: DBZOFD+TimesNewRomanPS-ItalicMT Encoding: 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Reyna-Llorens DOI: 10.1093/jxb/erag161; Journal of Experimental Botany, 77, 14, 2026-04-10.; Abstract: Plants have evolved carbon-concentrating mechanisms (CCMs) to optimize carbon fixation under environmental conditions that increase photorespiratory and transpiration rates. C4 and Crassulacean acid metabolism (CAM) photosynthesis are the two most widespread CCM examples across land plants and were typically considered to be incompatible due to biochemical, anatomical, evolutionary, and regulatory characteristics. However, Portulaca oleracea (purslane) was the first reported case to challenge this assumption >40 years ago. Although it remains one of the most characterized C4–CAM species, many aspects of this dual system are still unresolved. This expert view summarizes recent advances in understanding C4–CAM integration in Portulaca, highlighting key genomic, transcriptomic, biochemical, and physiological insights, and outlines the most pressing questions about the functioning and evolution of this combined CCM. Future directions for explaining how these convergent pathways emerged in the same lineage include functional studies on key regulatory elements, spatially resolved analyses for protein distribution and metabolic fluxes across leaves, an updated phylogeny of the family, and the integration of molecular and physiological data. So far, it is evident that expressing a weak CAM cycle in a C4 crop might be possible and could entail increased survival, possibly contributing to higher productivity in a future with climate change. Abiotic stress carbon-concentrating mechanism C4 photosynthesis Crassulacean acid metabolism drought stress facultative CAM PEPC phosphoenolpyruvate carboxylase photosynthesis Portulaca oleracea © The Author(s) 2026. Published by Oxford University Press on behalf of the Society for Experimental Biology. Journal Journal of Experimental Botany © The Author(s) 2026. 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