Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei
| Autor: | Katarzyna Juzoń, Veronika Koláčková, Michaela Sedlářová, Ondřej Novák, Martin Kubeš, Vladimír Skalický, Jan Vrána, Tereza Vojtková, Aleš Pěnčík |
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| Rok vydání: | 2021 |
| Předmět: |
Indoles
Arabidopsis Cell Culture Techniques Centrifugation Mass Spectrometry Plant Growth Regulators Homeostasis heterocyclic compounds Biology (General) Spectroscopy chemistry.chemical_classification biology Chemistry Protoplasts food and beverages General Medicine subcellular fractionation Computer Science Applications Cell biology medicine.anatomical_structure Cell fractionation QH301-705.5 Cell Fractionation Article Catalysis Inorganic Chemistry Auxin Plant Cells Tobacco Metabolome medicine Physical and Theoretical Chemistry auxin metabolism QD1-999 Molecular Biology Cell Nucleus Indoleacetic Acids Auxin homeostasis flow cytometry QH Endoplasmic reticulum nucleus QK fungi Organic Chemistry biology.organism_classification Cytosol auxin Nucleus |
| Zdroj: | International Journal of Molecular Sciences Volume 22 Issue 22 International Journal of Molecular Sciences, Vol 22, Iss 12369, p 12369 (2021) |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms222212369 |
| Popis: | The plant nucleus plays an irreplaceable role in cellular control and regulation by auxin (indole-3-acetic acid, IAA) mainly because canonical auxin signaling takes place here. Auxin can enter the nucleus from either the endoplasmic reticulum or cytosol. Therefore, new information about the auxin metabolome (auxinome) in the nucleus can illuminate our understanding of subcellular auxin homeostasis. Different methods of nuclear isolation from various plant tissues have been described previously, but information about auxin metabolite levels in nuclei is still fragmented and insufficient. Herein, we tested several published nucleus isolation protocols based on differential centrifugation or flow cytometry. The optimized sorting protocol leading to promising yield, intactness, and purity was then combined with an ultra-sensitive mass spectrometry analysis. Using this approach, we can present the first complex report on the auxinome of isolated nuclei from cell cultures of Arabidopsis and tobacco. Moreover, our results show dynamic changes in auxin homeostasis at the intranuclear level after treatment of protoplasts with free IAA, or indole as a precursor of auxin biosynthesis. Finally, we can conclude that the methodological procedure combining flow cytometry and mass spectrometry offers new horizons for the study of auxin homeostasis at the subcellular level. |
| Databáze: | OpenAIRE |
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