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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

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Official URL: https://doi.org/10.3390/ijms222212369

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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
Subjects: Q Science > QH Natural history
Q Science > QK Botany
Divisions: Faculty of 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:
DateEvent
16 November 2021Published
12 November 2021Accepted
Volume: 22
Number: 22
Article Number: e12369
DOI: 10.3390/ijms222212369
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Open Access
RIOXX Funder/Project Grant:
Project/Grant IDRIOXX Funder NameFunder ID
CZ.02.1.01/0.0/0.0/16_019/0000827Czech Republic.‏ ‎Ministerstvo školství, mládeže a tělovýchovyhttp://viaf.org/viaf/312885350
IGA_PrF_2021_011 Univerzita Palackého v Olomoucihttp://dx.doi.org/10.13039/501100007059
IGA_PrF_2021_016)Univerzita Palackého v Olomoucihttp://dx.doi.org/10.13039/501100007059
Endowment fund Univerzita Palackého v Olomoucihttp://dx.doi.org/10.13039/501100007059
MSCA-IF project CrysPINs (792329)European Commissionhttp://dx.doi.org/10.13039/501100000780
17-21581Y Grantová Agentura České Republiky UNSPECIFIED

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