A comet assay protocol for analysis of DNA damage in cryopreserved nuclei from Arabidopsis seeds, seedlings, and leaves
The comet assay is classically applied to study DNA damage at the single-cell level. In plants, it is commonly used for the analysis of seedlings, leaves, and seeds. The assay is time-consuming and runs continuously, making it difficult to process large volumes of samples or samples derived from kin...
| Main Authors: | , , |
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| Format: | Artículo |
| Language: | Inglés |
| Published: |
Elsevier
2025
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| Subjects: | |
| Online Access: | http://hdl.handle.net/20.500.12123/24551 https://www.sciencedirect.com/science/article/abs/pii/S0168945225004169?via%3Dihub https://doi.org/10.1016/j.plantsci.2025.112798 |
| _version_ | 1855487345063624704 |
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| author | Torres, José Roberto Lescano Lopez, Carlos Ignacio Alvarez, María Elena |
| author_browse | Alvarez, María Elena Lescano Lopez, Carlos Ignacio Torres, José Roberto |
| author_facet | Torres, José Roberto Lescano Lopez, Carlos Ignacio Alvarez, María Elena |
| author_sort | Torres, José Roberto |
| collection | INTA Digital |
| description | The comet assay is classically applied to study DNA damage at the single-cell level. In plants, it is commonly used for the analysis of seedlings, leaves, and seeds. The assay is time-consuming and runs continuously, making it difficult to process large volumes of samples or samples derived from kinetic studies in parallel. To overcome these limitations, we present an optimized protocol of neutral comet assay for analysis of Arabidopsis seeds, seedlings and leaves. It includes the cryopreservation of nuclei under conditions that do not affect DNA integrity. This option is not present in other available protocols for plants and allows samples collected at different times to be preserved and then processed simultaneously. We provide details on all experimental steps, including slide preparation, to obtain high-quality samples without the need of commercial kits. Moreover, we demonstrated its effectiveness in quantifying DNA damage induced by abiotic and biotic stresses. This versatile approach will help to optimize sample processing and will contribute to improving the studies of DNA damage in plants. |
| format | Artículo |
| id | INTA24551 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA245512025-11-13T12:37:44Z A comet assay protocol for analysis of DNA damage in cryopreserved nuclei from Arabidopsis seeds, seedlings, and leaves Torres, José Roberto Lescano Lopez, Carlos Ignacio Alvarez, María Elena Cryopreservation Seeds Seedling Leaves DNA Repair Criopreservación Semillas Plántulas Hojas Reparación de ADN Arabidopsis Comet Assay Frozen Nuclei Nuclear DNA Damage DNA Strand Breaks The comet assay is classically applied to study DNA damage at the single-cell level. In plants, it is commonly used for the analysis of seedlings, leaves, and seeds. The assay is time-consuming and runs continuously, making it difficult to process large volumes of samples or samples derived from kinetic studies in parallel. To overcome these limitations, we present an optimized protocol of neutral comet assay for analysis of Arabidopsis seeds, seedlings and leaves. It includes the cryopreservation of nuclei under conditions that do not affect DNA integrity. This option is not present in other available protocols for plants and allows samples collected at different times to be preserved and then processed simultaneously. We provide details on all experimental steps, including slide preparation, to obtain high-quality samples without the need of commercial kits. Moreover, we demonstrated its effectiveness in quantifying DNA damage induced by abiotic and biotic stresses. This versatile approach will help to optimize sample processing and will contribute to improving the studies of DNA damage in plants. Instituto de Fisiología y Recursos Genéticos Vegetales Fil: Torres, José Roberto. Universidad Nacional de Córdoba (UNC). Facultad de Ciencias Químicas. Departamento de Química Biológica Ranwel Caputto. Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC); Argentina Fil: Torres, José Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC); Argentina Fil: Lescano Lopez, Carlos Ignacio. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Fisiología y Recursos Genéticos Vegetales; Argentina Fil: Lescano Lopez, Carlos Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Estudios Agropecuarios (UDEA); Argentina Fil: Alvarez, María Elena. Universidad Nacional de Córdoba (UNC). Facultad de Ciencias Químicas. Departamento de Química Biológica Ranwel Caputto. Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC); Argentina Fil: Alvarez, María Elena. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC); Argentina 2025-11-13T12:20:33Z 2025-11-13T12:20:33Z 2025-12 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/24551 https://www.sciencedirect.com/science/article/abs/pii/S0168945225004169?via%3Dihub 0168-9452 https://doi.org/10.1016/j.plantsci.2025.112798 eng info:eu-repo/semantics/restrictedAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Elsevier Plant Science 361 : 112798 (December 2025) |
| spellingShingle | Cryopreservation Seeds Seedling Leaves DNA Repair Criopreservación Semillas Plántulas Hojas Reparación de ADN Arabidopsis Comet Assay Frozen Nuclei Nuclear DNA Damage DNA Strand Breaks Torres, José Roberto Lescano Lopez, Carlos Ignacio Alvarez, María Elena A comet assay protocol for analysis of DNA damage in cryopreserved nuclei from Arabidopsis seeds, seedlings, and leaves |
| title | A comet assay protocol for analysis of DNA damage in cryopreserved nuclei from Arabidopsis seeds, seedlings, and leaves |
| title_full | A comet assay protocol for analysis of DNA damage in cryopreserved nuclei from Arabidopsis seeds, seedlings, and leaves |
| title_fullStr | A comet assay protocol for analysis of DNA damage in cryopreserved nuclei from Arabidopsis seeds, seedlings, and leaves |
| title_full_unstemmed | A comet assay protocol for analysis of DNA damage in cryopreserved nuclei from Arabidopsis seeds, seedlings, and leaves |
| title_short | A comet assay protocol for analysis of DNA damage in cryopreserved nuclei from Arabidopsis seeds, seedlings, and leaves |
| title_sort | comet assay protocol for analysis of dna damage in cryopreserved nuclei from arabidopsis seeds seedlings and leaves |
| topic | Cryopreservation Seeds Seedling Leaves DNA Repair Criopreservación Semillas Plántulas Hojas Reparación de ADN Arabidopsis Comet Assay Frozen Nuclei Nuclear DNA Damage DNA Strand Breaks |
| url | http://hdl.handle.net/20.500.12123/24551 https://www.sciencedirect.com/science/article/abs/pii/S0168945225004169?via%3Dihub https://doi.org/10.1016/j.plantsci.2025.112798 |
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