Cargando…

Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei

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 me...

Descripción completa

Detalles Bibliográficos
Autores principales: Skalický, Vladimír, Vojtková, Tereza, Pěnčík, Aleš, Vrána, Jan, Juzoń, Katarzyna, Koláčková, Veronika, Sedlářová, Michaela, Kubeš, Martin F., Novák, Ondřej
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8620152/
https://www.ncbi.nlm.nih.gov/pubmed/34830250
http://dx.doi.org/10.3390/ijms222212369
_version_ 1784605153353007104
author Skalický, Vladimír
Vojtková, Tereza
Pěnčík, Aleš
Vrána, Jan
Juzoń, Katarzyna
Koláčková, Veronika
Sedlářová, Michaela
Kubeš, Martin F.
Novák, Ondřej
author_facet Skalický, Vladimír
Vojtková, Tereza
Pěnčík, Aleš
Vrána, Jan
Juzoń, Katarzyna
Koláčková, Veronika
Sedlářová, Michaela
Kubeš, Martin F.
Novák, Ondřej
author_sort Skalický, Vladimír
collection PubMed
description 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.
format Online
Article
Text
id pubmed-8620152
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-86201522021-11-27 Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei Skalický, Vladimír Vojtková, Tereza Pěnčík, Aleš Vrána, Jan Juzoń, Katarzyna Koláčková, Veronika Sedlářová, Michaela Kubeš, Martin F. Novák, Ondřej Int J Mol Sci Article 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. MDPI 2021-11-16 /pmc/articles/PMC8620152/ /pubmed/34830250 http://dx.doi.org/10.3390/ijms222212369 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Skalický, Vladimír
Vojtková, Tereza
Pěnčík, Aleš
Vrána, Jan
Juzoń, Katarzyna
Koláčková, Veronika
Sedlářová, Michaela
Kubeš, Martin F.
Novák, Ondřej
Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei
title Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei
title_full Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei
title_fullStr Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei
title_full_unstemmed Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei
title_short Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei
title_sort auxin metabolite profiling in isolated and intact plant nuclei
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8620152/
https://www.ncbi.nlm.nih.gov/pubmed/34830250
http://dx.doi.org/10.3390/ijms222212369
work_keys_str_mv AT skalickyvladimir auxinmetaboliteprofilinginisolatedandintactplantnuclei
AT vojtkovatereza auxinmetaboliteprofilinginisolatedandintactplantnuclei
AT pencikales auxinmetaboliteprofilinginisolatedandintactplantnuclei
AT vranajan auxinmetaboliteprofilinginisolatedandintactplantnuclei
AT juzonkatarzyna auxinmetaboliteprofilinginisolatedandintactplantnuclei
AT kolackovaveronika auxinmetaboliteprofilinginisolatedandintactplantnuclei
AT sedlarovamichaela auxinmetaboliteprofilinginisolatedandintactplantnuclei
AT kubesmartinf auxinmetaboliteprofilinginisolatedandintactplantnuclei
AT novakondrej auxinmetaboliteprofilinginisolatedandintactplantnuclei