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Hunting monolignol transporters: membrane proteomics and biochemical transport assays with membrane vesicles of Norway spruce

Both the mechanisms of monolignol transport and the transported form of monolignols in developing xylem of trees are unknown. We tested the hypothesis of an active, plasma membrane-localized transport of monolignol monomers, dimers, and/or glucosidic forms with membrane vesicles prepared from develo...

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Autores principales: Väisänen, Enni, Takahashi, Junko, Obudulu, Ogonna, Bygdell, Joakim, Karhunen, Pirkko, Blokhina, Olga, Laitinen, Teresa, Teeri, Teemu H, Wingsle, Gunnar, Fagerstedt, Kurt V, Kärkönen, Anna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7586744/
https://www.ncbi.nlm.nih.gov/pubmed/32777074
http://dx.doi.org/10.1093/jxb/eraa368
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author Väisänen, Enni
Takahashi, Junko
Obudulu, Ogonna
Bygdell, Joakim
Karhunen, Pirkko
Blokhina, Olga
Laitinen, Teresa
Teeri, Teemu H
Wingsle, Gunnar
Fagerstedt, Kurt V
Kärkönen, Anna
author_facet Väisänen, Enni
Takahashi, Junko
Obudulu, Ogonna
Bygdell, Joakim
Karhunen, Pirkko
Blokhina, Olga
Laitinen, Teresa
Teeri, Teemu H
Wingsle, Gunnar
Fagerstedt, Kurt V
Kärkönen, Anna
author_sort Väisänen, Enni
collection PubMed
description Both the mechanisms of monolignol transport and the transported form of monolignols in developing xylem of trees are unknown. We tested the hypothesis of an active, plasma membrane-localized transport of monolignol monomers, dimers, and/or glucosidic forms with membrane vesicles prepared from developing xylem and lignin-forming tissue-cultured cells of Norway spruce (Picea abies L. Karst.), as well as from control materials, comprising non-lignifying Norway spruce phloem and tobacco (Nicotiana tabacum L.) BY-2 cells. Xylem and BY-2 vesicles transported both coniferin and p-coumaryl alcohol glucoside, but inhibitor assays suggested that this transport was through the tonoplast. Membrane vesicles prepared from lignin-forming spruce cells showed coniferin transport, but the K(m) value for coniferin was much higher than those of xylem and BY-2 cells. Liquid chromatography-mass spectrometry analysis of membrane proteins isolated from spruce developing xylem, phloem, and lignin-forming cultured cells revealed multiple transporters. These were compared with a transporter gene set obtained by a correlation analysis with a selected set of spruce monolignol biosynthesis genes. Biochemical membrane vesicle assays showed no support for ABC-transporter-mediated monolignol transport but point to a role for secondary active transporters (such as MFS or MATE transporters). In contrast, proteomic and co-expression analyses suggested a role for ABC transporters and MFS transporters.
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spelling pubmed-75867442020-10-30 Hunting monolignol transporters: membrane proteomics and biochemical transport assays with membrane vesicles of Norway spruce Väisänen, Enni Takahashi, Junko Obudulu, Ogonna Bygdell, Joakim Karhunen, Pirkko Blokhina, Olga Laitinen, Teresa Teeri, Teemu H Wingsle, Gunnar Fagerstedt, Kurt V Kärkönen, Anna J Exp Bot Research Papers Both the mechanisms of monolignol transport and the transported form of monolignols in developing xylem of trees are unknown. We tested the hypothesis of an active, plasma membrane-localized transport of monolignol monomers, dimers, and/or glucosidic forms with membrane vesicles prepared from developing xylem and lignin-forming tissue-cultured cells of Norway spruce (Picea abies L. Karst.), as well as from control materials, comprising non-lignifying Norway spruce phloem and tobacco (Nicotiana tabacum L.) BY-2 cells. Xylem and BY-2 vesicles transported both coniferin and p-coumaryl alcohol glucoside, but inhibitor assays suggested that this transport was through the tonoplast. Membrane vesicles prepared from lignin-forming spruce cells showed coniferin transport, but the K(m) value for coniferin was much higher than those of xylem and BY-2 cells. Liquid chromatography-mass spectrometry analysis of membrane proteins isolated from spruce developing xylem, phloem, and lignin-forming cultured cells revealed multiple transporters. These were compared with a transporter gene set obtained by a correlation analysis with a selected set of spruce monolignol biosynthesis genes. Biochemical membrane vesicle assays showed no support for ABC-transporter-mediated monolignol transport but point to a role for secondary active transporters (such as MFS or MATE transporters). In contrast, proteomic and co-expression analyses suggested a role for ABC transporters and MFS transporters. Oxford University Press 2020-08-10 /pmc/articles/PMC7586744/ /pubmed/32777074 http://dx.doi.org/10.1093/jxb/eraa368 Text en © The Author(s) 2020. Published by Oxford University Press on behalf of the Society for Experimental Biology. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Papers
Väisänen, Enni
Takahashi, Junko
Obudulu, Ogonna
Bygdell, Joakim
Karhunen, Pirkko
Blokhina, Olga
Laitinen, Teresa
Teeri, Teemu H
Wingsle, Gunnar
Fagerstedt, Kurt V
Kärkönen, Anna
Hunting monolignol transporters: membrane proteomics and biochemical transport assays with membrane vesicles of Norway spruce
title Hunting monolignol transporters: membrane proteomics and biochemical transport assays with membrane vesicles of Norway spruce
title_full Hunting monolignol transporters: membrane proteomics and biochemical transport assays with membrane vesicles of Norway spruce
title_fullStr Hunting monolignol transporters: membrane proteomics and biochemical transport assays with membrane vesicles of Norway spruce
title_full_unstemmed Hunting monolignol transporters: membrane proteomics and biochemical transport assays with membrane vesicles of Norway spruce
title_short Hunting monolignol transporters: membrane proteomics and biochemical transport assays with membrane vesicles of Norway spruce
title_sort hunting monolignol transporters: membrane proteomics and biochemical transport assays with membrane vesicles of norway spruce
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7586744/
https://www.ncbi.nlm.nih.gov/pubmed/32777074
http://dx.doi.org/10.1093/jxb/eraa368
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