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Article

Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei

1
Laboratory of Growth Regulators, Institute of Experimental Botany of the Czech Academy of Sciences and Faculty of Science, Palacký University, Šlechtitelů 27, CZ-78371 Olomouc, Czech Republic
2
Institute of Experimental Botany of the Czech Academy of Sciences, Centre of the Region Haná for Biotechnological and Agricultural Research, Šlechtitelů 31, CZ-77900 Olomouc, Czech Republic
3
Department of Biotechnology, The Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Niezapominajek 21, 30-239 Krakow, Poland
4
Department of Botany, Faculty of Science, Palacký University, Šlechtitelů 27, CZ-78371 Olomouc, Czech Republic
5
School of Life Sciences, University of Warwick, Coventry CV4 7AL, UK
*
Authors to whom correspondence should be addressed.
Current Address: Department of Biology, Faculty of Science, University of Hradec Králové, Rokitanského 62, CZ-50003 Hradec Králové, Czech Republic.
Academic Editor: Massimo Maffei
Int. J. Mol. Sci. 2021, 22(22), 12369; https://doi.org/10.3390/ijms222212369
Received: 8 October 2021 / Revised: 11 November 2021 / Accepted: 12 November 2021 / Published: 16 November 2021
(This article belongs to the Special Issue Growth Regulators in Plants)
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. View Full-Text
Keywords: subcellular fractionation; flow cytometry; nucleus; auxin; auxin metabolism subcellular fractionation; flow cytometry; nucleus; auxin; auxin metabolism
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MDPI and ACS Style

Skalický, V.; Vojtková, T.; Pěnčík, A.; Vrána, J.; Juzoń, K.; Koláčková, V.; Sedlářová, M.; Kubeš, M.F.; Novák, O. Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei. Int. J. Mol. Sci. 2021, 22, 12369. https://doi.org/10.3390/ijms222212369

AMA Style

Skalický V, Vojtková T, Pěnčík A, Vrána J, Juzoń K, Koláčková V, Sedlářová M, Kubeš MF, Novák O. Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei. International Journal of Molecular Sciences. 2021; 22(22):12369. https://doi.org/10.3390/ijms222212369

Chicago/Turabian Style

Skalický, Vladimír, Tereza Vojtková, Aleš Pěnčík, Jan Vrána, Katarzyna Juzoń, Veronika Koláčková, Michaela Sedlářová, Martin F. Kubeš, and Ondřej Novák. 2021. "Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei" International Journal of Molecular Sciences 22, no. 22: 12369. https://doi.org/10.3390/ijms222212369

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