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Auxin metabolite profiling in isolated and intact plant nuclei
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Skalický, Vladimír, Vojtková, Tereza, Pěnčík, Aleš, Vrána, Jan, Juzoń, Katarzyna, Koláčková, Veronika, Sedlářová, Michaela, Kubeš, Martin F. and Novák, Ondřej (2021) Auxin metabolite profiling in isolated and intact plant nuclei. International Journal of Molecular Sciences, 22 (22). e12369. doi:10.3390/ijms222212369 ISSN 1422-0067.
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WRAP-Auxin-metabolite-profiling-isolated-intact-plant-nuclei-Kubes-2021.pdf - Published Version - Requires a PDF viewer. Available under License Creative Commons Attribution 4.0. Download (2375Kb) | Preview |
Official URL: https://doi.org/10.3390/ijms222212369
Abstract
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.
Item Type: | Journal Article | |||||||||||||||||||||
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Subjects: | Q Science > QH Natural history Q Science > QK Botany |
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | |||||||||||||||||||||
SWORD Depositor: | Library Publications Router | |||||||||||||||||||||
Library of Congress Subject Headings (LCSH): | Subcellular fractionation , Flow cytometry , Plant cells and tissues , Auxin , Auxin -- Molecular aspects | |||||||||||||||||||||
Journal or Publication Title: | International Journal of Molecular Sciences | |||||||||||||||||||||
Publisher: | M D P I AG | |||||||||||||||||||||
ISSN: | 1422-0067 | |||||||||||||||||||||
Official Date: | 16 November 2021 | |||||||||||||||||||||
Dates: |
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Volume: | 22 | |||||||||||||||||||||
Number: | 22 | |||||||||||||||||||||
Article Number: | e12369 | |||||||||||||||||||||
DOI: | 10.3390/ijms222212369 | |||||||||||||||||||||
Status: | Peer Reviewed | |||||||||||||||||||||
Publication Status: | Published | |||||||||||||||||||||
Access rights to Published version: | Open Access (Creative Commons) | |||||||||||||||||||||
Date of first compliant deposit: | 29 November 2021 | |||||||||||||||||||||
Date of first compliant Open Access: | 2 December 2021 | |||||||||||||||||||||
RIOXX Funder/Project Grant: |
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