A simple and highly efficient protocol for 13C-labeling of plant cell wall for structural and quantitative analyses via solid-state nuclear magnetic resonance
Abstract Background Plant cell walls are made of a complex network of interacting polymers that play a critical role in plant development and responses to environmental changes. Thus, improving plant biomass and fitness requires the elucidation of the structural organization of plant cell walls in t...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2025-01-01
|
| Series: | Plant Methods |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13007-024-01310-3 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832594731187568640 |
|---|---|
| author | Tasleem Javaid Akshayaa Venkataraghavan Matrika Bhattarai Debkumar Debnath Wancheng Zhao Tuo Wang Ahmed Faik |
| author_facet | Tasleem Javaid Akshayaa Venkataraghavan Matrika Bhattarai Debkumar Debnath Wancheng Zhao Tuo Wang Ahmed Faik |
| author_sort | Tasleem Javaid |
| collection | DOAJ |
| description | Abstract Background Plant cell walls are made of a complex network of interacting polymers that play a critical role in plant development and responses to environmental changes. Thus, improving plant biomass and fitness requires the elucidation of the structural organization of plant cell walls in their native environment. The 13C-based multi-dimensional solid-state nuclear magnetic resonance (ssNMR) has been instrumental in revealing the structural information of plant cell walls through 2D and 3D correlation spectral analyses. However, the requirement of enriching plants with 13C limits the applicability of this method. To our knowledge, there is only a very limited set of methods currently available that achieve high levels of 13C-labeling of plant materials using 13CO2, and most of them require large amounts of 13CO2 in larger growth chambers. Results In this study, a simplified protocol for 13C-labeling of plant materials is introduced that allows ca 60% labeling of the cell walls, as quantified by comparison with commercially labeled samples. This level of 13C-enrichment is sufficient for all conventional 2D and 3D correlation ssNMR experiments for detailed analysis of plant cell wall structure. The protocol is based on a convenient and easy setup to supply both 13C-labeled glucose and 13CO2 using a vacuum-desiccator. The protocol does not require large amounts of 13CO2. Conclusion This study shows that our 13C-labeling of plant materials can make the accessibility to ssNMR technique easy and affordable. The derived high-resolution 2D and 3D correlation spectra are used to extract structural information of plant cell walls. This helps to better understand the influence of polysaccharide-polysaccharide interaction on plant performance and allows for a more precise parametrization of plant cell wall models. |
| format | Article |
| id | doaj-art-f1a5389ca7c24788950a32a925eafe33 |
| institution | Kabale University |
| issn | 1746-4811 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Plant Methods |
| spelling | doaj-art-f1a5389ca7c24788950a32a925eafe332025-01-19T12:25:13ZengBMCPlant Methods1746-48112025-01-012111810.1186/s13007-024-01310-3A simple and highly efficient protocol for 13C-labeling of plant cell wall for structural and quantitative analyses via solid-state nuclear magnetic resonanceTasleem Javaid0Akshayaa Venkataraghavan1Matrika Bhattarai2Debkumar Debnath3Wancheng Zhao4Tuo Wang5Ahmed Faik6Department of Environmental and Plant Biology, Ohio UniversityDepartment of Environmental and Plant Biology, Ohio UniversityDepartment of Environmental and Plant Biology, Ohio UniversityThe Department of Chemistry, Michigan State UniversityThe Department of Chemistry, Michigan State UniversityThe Department of Chemistry, Michigan State UniversityDepartment of Environmental and Plant Biology, Ohio UniversityAbstract Background Plant cell walls are made of a complex network of interacting polymers that play a critical role in plant development and responses to environmental changes. Thus, improving plant biomass and fitness requires the elucidation of the structural organization of plant cell walls in their native environment. The 13C-based multi-dimensional solid-state nuclear magnetic resonance (ssNMR) has been instrumental in revealing the structural information of plant cell walls through 2D and 3D correlation spectral analyses. However, the requirement of enriching plants with 13C limits the applicability of this method. To our knowledge, there is only a very limited set of methods currently available that achieve high levels of 13C-labeling of plant materials using 13CO2, and most of them require large amounts of 13CO2 in larger growth chambers. Results In this study, a simplified protocol for 13C-labeling of plant materials is introduced that allows ca 60% labeling of the cell walls, as quantified by comparison with commercially labeled samples. This level of 13C-enrichment is sufficient for all conventional 2D and 3D correlation ssNMR experiments for detailed analysis of plant cell wall structure. The protocol is based on a convenient and easy setup to supply both 13C-labeled glucose and 13CO2 using a vacuum-desiccator. The protocol does not require large amounts of 13CO2. Conclusion This study shows that our 13C-labeling of plant materials can make the accessibility to ssNMR technique easy and affordable. The derived high-resolution 2D and 3D correlation spectra are used to extract structural information of plant cell walls. This helps to better understand the influence of polysaccharide-polysaccharide interaction on plant performance and allows for a more precise parametrization of plant cell wall models.https://doi.org/10.1186/s13007-024-01310-3Stable isotope labelingPlant cell wallSolid-state NMR (ssNMR)RicePolysaccharides |
| spellingShingle | Tasleem Javaid Akshayaa Venkataraghavan Matrika Bhattarai Debkumar Debnath Wancheng Zhao Tuo Wang Ahmed Faik A simple and highly efficient protocol for 13C-labeling of plant cell wall for structural and quantitative analyses via solid-state nuclear magnetic resonance Plant Methods Stable isotope labeling Plant cell wall Solid-state NMR (ssNMR) Rice Polysaccharides |
| title | A simple and highly efficient protocol for 13C-labeling of plant cell wall for structural and quantitative analyses via solid-state nuclear magnetic resonance |
| title_full | A simple and highly efficient protocol for 13C-labeling of plant cell wall for structural and quantitative analyses via solid-state nuclear magnetic resonance |
| title_fullStr | A simple and highly efficient protocol for 13C-labeling of plant cell wall for structural and quantitative analyses via solid-state nuclear magnetic resonance |
| title_full_unstemmed | A simple and highly efficient protocol for 13C-labeling of plant cell wall for structural and quantitative analyses via solid-state nuclear magnetic resonance |
| title_short | A simple and highly efficient protocol for 13C-labeling of plant cell wall for structural and quantitative analyses via solid-state nuclear magnetic resonance |
| title_sort | simple and highly efficient protocol for 13c labeling of plant cell wall for structural and quantitative analyses via solid state nuclear magnetic resonance |
| topic | Stable isotope labeling Plant cell wall Solid-state NMR (ssNMR) Rice Polysaccharides |
| url | https://doi.org/10.1186/s13007-024-01310-3 |
| work_keys_str_mv | AT tasleemjavaid asimpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance AT akshayaavenkataraghavan asimpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance AT matrikabhattarai asimpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance AT debkumardebnath asimpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance AT wanchengzhao asimpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance AT tuowang asimpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance AT ahmedfaik asimpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance AT tasleemjavaid simpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance AT akshayaavenkataraghavan simpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance AT matrikabhattarai simpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance AT debkumardebnath simpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance AT wanchengzhao simpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance AT tuowang simpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance AT ahmedfaik simpleandhighlyefficientprotocolfor13clabelingofplantcellwallforstructuralandquantitativeanalysesviasolidstatenuclearmagneticresonance |