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      Phellem Cell-Wall Components Are Discriminants of Cork Quality in Quercus suber

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          Abstract

          Cork is a renewable, non-wood high valued forest product, with relevant ecological and economic impact in the Mediterranean-type ecosystems. Currently, cork is ranked according to its commercial quality. The most valuable planks are chosen for cork stoppers production. Cork planks with adequate thickness and porosity are classified as stoppable quality cork (SQC). The chemical composition of cork is known, but the regulation of metabolic pathways responsible of cork production and composition, hence of cork quality, is largely unknown. Here, we tested the hypothesis that post-genomic events may be responsible for the development of SQC and N-SQC (non-stoppable quality cork). Here, we show that combined proteomics and targeted metabolomics (namely soluble and cell wall bound phenolics) analyzed on recently formed phellem allows discriminate cork planks of different quality. Phellem cells of SQC and N-SQC displayed different reducing capacity, with consequential impact on both enzymatic pathways (e.g., glycolysis) and other cellular functions, including cell wall assembly and suberization. Glycolysis and respiration related proteins were abundant in both cork quality groups, whereas the level of several proteins associated to mitochondrial metabolism was higher in N-SQC. The soluble and cell wall-bound phenolics in recently formed phellem clearly discriminated SQC from N-SCQ. In our study, SQC was characterized by a high incorporation of aromatic components of the phenylpropanoid pathway in the cell wall, together with a lower content of hydrolysable tannins. Here, we propose that the level of hydrolysable tannins may represent a valuable diagnostic tool for screening recently formed phellem, and used as a proxy for the quality grade of cork plank produced by each tree.

          Graphical Abstract

          The shikimate and the core phenylpropanoid pathways are involved in the synthesis of the polyphenols identified in cork producing cells. Stoppable cork quality producing cells are characterized by higher incorporation of aromatic components of the phenylpropanoid pathway in the cell wall, together with a lower content of hydrolysable tannins.

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          Most cited references36

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          Techniques for Analysis of Plant Phenolic Compounds

          Phenolic compounds are well-known phytochemicals found in all plants. They consist of simple phenols, benzoic and cinnamic acid, coumarins, tannins, lignins, lignans and flavonoids. Substantial developments in research focused on the extraction, identification and quantification of phenolic compounds as medicinal and/or dietary molecules have occurred over the last 25 years. Organic solvent extraction is the main method used to extract phenolics. Chemical procedures are used to detect the presence of total phenolics, while spectrophotometric and chromatographic techniques are utilized to identify and quantify individual phenolic compounds. This review addresses the application of different methodologies utilized in the analysis of phenolic compounds in plant-based products, including recent technical developments in the quantification of phenolics.
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            Sample extraction techniques for enhanced proteomic analysis of plant tissues.

            Major improvements in proteomic techniques in recent years have led to an increase in their application in all biological fields, including plant sciences. For all proteomic approaches, protein extraction and sample preparation are of utmost importance for optimal results; however, extraction of proteins from plant tissues represents a great challenge. Plant tissues usually contain relatively low amounts of proteins and high concentrations of proteases and compounds that potentially can limit tissue disintegration and interfere with subsequent protein separation and identification. An effective protein extraction protocol must also be adaptable to the great variation in the sets of secondary metabolites and potentially contaminating compounds that occurs between tissues (e.g., leaves, roots, fruit, seeds and stems) and between species. Here we present two basic protein extraction protocols that have successfully been used with diverse plant tissues, including recalcitrant tissues. The first method is based on phenol extraction coupled with ammonium acetate precipitation, and the second is based on trichloroacetic acid (TCA) precipitation. Both extraction protocols can be completed within 2 d.
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              Phenylpropanoid biosynthesis and its regulation.

              In the past year progress has been made in the manipulation of phenylpropanoid metabolism but several studies highlight gaps in our understanding of the biochemistry of these pathways. New components involved in transcriptional regulation of phenylpropanoid genes have been identified, including transcription factors and novel proteins that function upstream of DNA-binding proteins.
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                Author and article information

                Contributors
                Journal
                Front Plant Sci
                Front Plant Sci
                Front. Plant Sci.
                Frontiers in Plant Science
                Frontiers Media S.A.
                1664-462X
                30 July 2019
                2019
                : 10
                : 944
                Affiliations
                [1] 1Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa , Lisbon, Portugal
                [2] 2Instituto de Tecnologia Química e Biológica, Universidade NOVA de Lisboa , Lisbon, Portugal
                [3] 3Department of Ecogenomics and Systems Biology, University of Vienna , Vienna, Austria
                [4] 4UCIBIO – REQUIMTE, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa , Caparica, Portugal
                [5] 5National Research Council of Italy, Trees and Timber Institute , Florence, Italy
                [6] 6Department of Agri-Food Production and Environmental Sciences, University of Florence , Florence, Italy
                [7] 7Luxembourg Institute of Science and Technology , Belvaux, Luxembourg
                [8] 8Institute for Sustainable Plant Protection, National Research Council of Italy , Florence, Italy
                [9] 9Instituto Superior de Agronomia, Universidade de Lisboa , Lisbon, Portugal
                Author notes

                Edited by: Soren K. Rasmussen, University of Copenhagen, Denmark

                Reviewed by: Marco Landi, University of Pisa, Italy; Patricia C. Ferreira Dos Santos, University of Nevada, Reno, United States

                Present address: Rita Teresa Teixeira, Faculdade de Ciências de Lisboa, BioISI, Universidade de Lisboa, Lisbon, Portugal

                This article was submitted to Plant Breeding, a section of the journal Frontiers in Plant Science

                Article
                10.3389/fpls.2019.00944
                6682605
                d54cf498-235c-406b-a3db-760b8290d48e
                Copyright © 2019 Pinheiro, Wienkoop, de Almeida, Brunetti, Zarrouk, Planchon, Gori, Tattini, Ricardo, Renaut and Teixeira.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 02 April 2019
                : 08 July 2019
                Page count
                Figures: 7, Tables: 2, Equations: 0, References: 56, Pages: 15, Words: 0
                Funding
                Funded by: Fundação para a Ciência e a Tecnologia 10.13039/501100001871
                Award ID: PTDC/AGRAAM/100465/2008
                Funded by: Fonds National de la Recherche Luxembourg 10.13039/501100001866
                Categories
                Plant Science
                Original Research

                Plant science & Botany
                hydrolysable tannins,proteomics,targeted metabolomics,soluble phenolics,cell-wall immobilized phenolics

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