The Q‐Warg Pipeline: A Robust and Versatile Workflow for Quantitative Analysis of Protoplast Culture Conditions
ABSTRACT Single cells offer a simplified model for investigating complex mechanisms such as cell–cell adhesion. Protoplasts, plant cells without cell walls (CWs), have been instrumental in plant research, industrial applications, and breeding. However, because of the absence of a CW, protoplasts are...
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| Format: | Article |
| Language: | English |
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Wiley
2025-07-01
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| Series: | Plant Direct |
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| Online Access: | https://doi.org/10.1002/pld3.70090 |
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| author | Léa Bogdziewiez Rik Froeling Patricia Schöppl Jeanne Juquel Ioanna Antoniadi Vladimìr Skalický Ambroise Mathey Jacques Fattaccioli Joris Sprakel Stéphane Verger |
| author_facet | Léa Bogdziewiez Rik Froeling Patricia Schöppl Jeanne Juquel Ioanna Antoniadi Vladimìr Skalický Ambroise Mathey Jacques Fattaccioli Joris Sprakel Stéphane Verger |
| author_sort | Léa Bogdziewiez |
| collection | DOAJ |
| description | ABSTRACT Single cells offer a simplified model for investigating complex mechanisms such as cell–cell adhesion. Protoplasts, plant cells without cell walls (CWs), have been instrumental in plant research, industrial applications, and breeding. However, because of the absence of a CW, protoplasts are not considered “true” plant cells, making them less relevant for biophysical studies. Current protocols for CW recovery in protoplasts vary widely among laboratories and starting materials, requiring lab‐specific optimizations that often depend on expert knowledge and qualitative assessments. To address this, we have developed a user‐friendly streamlined workflow, the Q‐Warg pipeline, which enables quantitative comparison of various conditions for CW recovery post‐protoplasting. This pipeline employs fluorescence imaging and tailored processing to measure parameters such as morphometry, cell viability, and CW staining intensity. Using this approach, we optimized culture conditions to obtain single plant cells (SPCs) with recovered CWs. Additionally, we demonstrated the robustness and versatility of the workflow by quantifying different fluorescent signals in protoplast suspensions. Overall, the Q‐Warg pipeline provides a widely accessible and user‐friendly solution for robust and unbiased characterization of protoplasts culture. The quantitative data generated by the pipeline will be useful in the future to decipher the mechanisms regulating protoplast viability and regeneration. |
| format | Article |
| id | doaj-art-d8f9131000c24a8eb95e7f65a7e515b5 |
| institution | Kabale University |
| issn | 2475-4455 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Wiley |
| record_format | Article |
| series | Plant Direct |
| spelling | doaj-art-d8f9131000c24a8eb95e7f65a7e515b52025-08-20T03:31:37ZengWileyPlant Direct2475-44552025-07-0197n/an/a10.1002/pld3.70090The Q‐Warg Pipeline: A Robust and Versatile Workflow for Quantitative Analysis of Protoplast Culture ConditionsLéa Bogdziewiez0Rik Froeling1Patricia Schöppl2Jeanne Juquel3Ioanna Antoniadi4Vladimìr Skalický5Ambroise Mathey6Jacques Fattaccioli7Joris Sprakel8Stéphane Verger9Umeå Plant Science Centre (UPSC), Department of Forest Genetics and Plant Physiology Swedish University of Agricultural Sciences Umeå SwedenLaboratory of Biochemistry, Wageningen University and Research Wageningen the NetherlandsLaboratory of Biochemistry, Wageningen University and Research Wageningen the NetherlandsUmeå Plant Science Centre (UPSC), Department of Plant Physiology Umeå University Umeå SwedenUmeå Plant Science Centre (UPSC), Department of Forest Genetics and Plant Physiology Swedish University of Agricultural Sciences Umeå SwedenUmeå Plant Science Centre (UPSC), Department of Forest Genetics and Plant Physiology Swedish University of Agricultural Sciences Umeå SwedenUmeå Plant Science Centre (UPSC), Department of Forest Genetics and Plant Physiology Swedish University of Agricultural Sciences Umeå SwedenPasteur, Département de Chimie Ecole Normale Supérieure, PSL Université, Sorbonne université, CNRS Paris FranceLaboratory of Biochemistry, Wageningen University and Research Wageningen the NetherlandsUmeå Plant Science Centre (UPSC), Department of Forest Genetics and Plant Physiology Swedish University of Agricultural Sciences Umeå SwedenABSTRACT Single cells offer a simplified model for investigating complex mechanisms such as cell–cell adhesion. Protoplasts, plant cells without cell walls (CWs), have been instrumental in plant research, industrial applications, and breeding. However, because of the absence of a CW, protoplasts are not considered “true” plant cells, making them less relevant for biophysical studies. Current protocols for CW recovery in protoplasts vary widely among laboratories and starting materials, requiring lab‐specific optimizations that often depend on expert knowledge and qualitative assessments. To address this, we have developed a user‐friendly streamlined workflow, the Q‐Warg pipeline, which enables quantitative comparison of various conditions for CW recovery post‐protoplasting. This pipeline employs fluorescence imaging and tailored processing to measure parameters such as morphometry, cell viability, and CW staining intensity. Using this approach, we optimized culture conditions to obtain single plant cells (SPCs) with recovered CWs. Additionally, we demonstrated the robustness and versatility of the workflow by quantifying different fluorescent signals in protoplast suspensions. Overall, the Q‐Warg pipeline provides a widely accessible and user‐friendly solution for robust and unbiased characterization of protoplasts culture. The quantitative data generated by the pipeline will be useful in the future to decipher the mechanisms regulating protoplast viability and regeneration.https://doi.org/10.1002/pld3.70090Arabidopsis thalianacell wallfluorescenceprotoplastsquantificationrecovery |
| spellingShingle | Léa Bogdziewiez Rik Froeling Patricia Schöppl Jeanne Juquel Ioanna Antoniadi Vladimìr Skalický Ambroise Mathey Jacques Fattaccioli Joris Sprakel Stéphane Verger The Q‐Warg Pipeline: A Robust and Versatile Workflow for Quantitative Analysis of Protoplast Culture Conditions Plant Direct Arabidopsis thaliana cell wall fluorescence protoplasts quantification recovery |
| title | The Q‐Warg Pipeline: A Robust and Versatile Workflow for Quantitative Analysis of Protoplast Culture Conditions |
| title_full | The Q‐Warg Pipeline: A Robust and Versatile Workflow for Quantitative Analysis of Protoplast Culture Conditions |
| title_fullStr | The Q‐Warg Pipeline: A Robust and Versatile Workflow for Quantitative Analysis of Protoplast Culture Conditions |
| title_full_unstemmed | The Q‐Warg Pipeline: A Robust and Versatile Workflow for Quantitative Analysis of Protoplast Culture Conditions |
| title_short | The Q‐Warg Pipeline: A Robust and Versatile Workflow for Quantitative Analysis of Protoplast Culture Conditions |
| title_sort | q warg pipeline a robust and versatile workflow for quantitative analysis of protoplast culture conditions |
| topic | Arabidopsis thaliana cell wall fluorescence protoplasts quantification recovery |
| url | https://doi.org/10.1002/pld3.70090 |
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