Immunolocalization of cyclotides in plant cells, tissues and organ supports their role in host defense
Identifieur interne : 000134 ( Pmc/Checkpoint ); précédent : 000133; suivant : 000135Immunolocalization of cyclotides in plant cells, tissues and organ supports their role in host defense
Auteurs : Blazej Slazak [Pologne, Suède] ; Małgorzata Kapusta [Pologne] ; Sohaib Malik [Suède] ; Jerzy Bohdanowicz [Pologne] ; El Bieta Kuta [Pologne] ; Przemysław Malec [Pologne] ; Ulf Göransson [Suède]Source :
- Planta [ 0032-0935 ] ; 2016.
Abstract
The cyclotide family of plant peptides is characterized by the cyclic cystine knot motif and its diverse biological activities. Given their insecticidal and antimicrobial properties, the role of cyclotides
The online version of this article (doi:10.1007/s00425-016-2562-y) contains supplementary material, which is available to authorized users.
Url:
DOI: 10.1007/s00425-016-2562-y
PubMed: 27394154
PubMed Central: 5052299
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Szafer Institute of Botany, Polish Academy of Science, 46 Lubicz St, 31-512 Cracow</wicri:regionArea><wicri:noRegion>31-512 Cracow</wicri:noRegion></affiliation><affiliation wicri:level="4"><nlm:aff id="Aff2">Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Center, Box 574, 751 23 Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Center, Box 574, 751 23 Uppsala</wicri:regionArea><orgName type="university">Université d'Uppsala</orgName><placeName><settlement type="city">Uppsala</settlement><region nuts="1">Svealand</region><region nuts="1">East Middle Sweden</region></placeName></affiliation></author><author><name sortKey="Kapusta, Malgorzata" sort="Kapusta, Malgorzata" uniqKey="Kapusta M" first="Małgorzata" last="Kapusta">Małgorzata Kapusta</name><affiliation wicri:level="1"><nlm:aff id="Aff3">Department of Plant Cytology and Embryology, Faculty of Biology, University of Gdańsk, 59 Wita Stwosza St, 80-308 Gdańsk, Poland</nlm:aff><country xml:lang="fr">Pologne</country><wicri:regionArea>Department of Plant Cytology and Embryology, Faculty of Biology, University of Gdańsk, 59 Wita Stwosza St, 80-308 Gdańsk</wicri:regionArea><wicri:noRegion>80-308 Gdańsk</wicri:noRegion></affiliation></author><author><name sortKey="Malik, Sohaib" sort="Malik, Sohaib" uniqKey="Malik S" first="Sohaib" last="Malik">Sohaib Malik</name><affiliation wicri:level="4"><nlm:aff id="Aff2">Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Center, Box 574, 751 23 Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Center, Box 574, 751 23 Uppsala</wicri:regionArea><orgName type="university">Université d'Uppsala</orgName><placeName><settlement type="city">Uppsala</settlement><region nuts="1">Svealand</region><region nuts="1">East Middle Sweden</region></placeName></affiliation></author><author><name sortKey="Bohdanowicz, Jerzy" sort="Bohdanowicz, Jerzy" uniqKey="Bohdanowicz J" first="Jerzy" last="Bohdanowicz">Jerzy Bohdanowicz</name><affiliation wicri:level="1"><nlm:aff id="Aff3">Department of Plant Cytology and Embryology, Faculty of Biology, University of Gdańsk, 59 Wita Stwosza St, 80-308 Gdańsk, Poland</nlm:aff><country xml:lang="fr">Pologne</country><wicri:regionArea>Department of Plant Cytology and Embryology, Faculty of Biology, University of Gdańsk, 59 Wita Stwosza St, 80-308 Gdańsk</wicri:regionArea><wicri:noRegion>80-308 Gdańsk</wicri:noRegion></affiliation></author><author><name sortKey="Kuta, El Bieta" sort="Kuta, El Bieta" uniqKey="Kuta E" first="El Bieta" last="Kuta">El Bieta Kuta</name><affiliation wicri:level="1"><nlm:aff id="Aff4">Department of Plant Cytology and Embryology, Institute of Botany, Jagiellonian University, 9 Gronostajowa St, 30-387 Cracow, Poland</nlm:aff><country xml:lang="fr">Pologne</country><wicri:regionArea>Department of Plant Cytology and Embryology, Institute of Botany, Jagiellonian University, 9 Gronostajowa St, 30-387 Cracow</wicri:regionArea><wicri:noRegion>30-387 Cracow</wicri:noRegion></affiliation></author><author><name sortKey="Malec, Przemyslaw" sort="Malec, Przemyslaw" uniqKey="Malec P" first="Przemysław" last="Malec">Przemysław Malec</name><affiliation wicri:level="1"><nlm:aff id="Aff5">Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 7 Gronostajowa St, 30-387 Cracow, Poland</nlm:aff><country xml:lang="fr">Pologne</country><wicri:regionArea>Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 7 Gronostajowa St, 30-387 Cracow</wicri:regionArea><wicri:noRegion>30-387 Cracow</wicri:noRegion></affiliation></author><author><name sortKey="Goransson, Ulf" sort="Goransson, Ulf" uniqKey="Goransson U" first="Ulf" last="Göransson">Ulf Göransson</name><affiliation wicri:level="4"><nlm:aff id="Aff2">Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Center, Box 574, 751 23 Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Center, Box 574, 751 23 Uppsala</wicri:regionArea><orgName type="university">Université d'Uppsala</orgName><placeName><settlement type="city">Uppsala</settlement><region nuts="1">Svealand</region><region nuts="1">East Middle Sweden</region></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">27394154</idno><idno type="pmc">5052299</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5052299</idno><idno type="RBID">PMC:5052299</idno><idno type="doi">10.1007/s00425-016-2562-y</idno><date when="2016">2016</date><idno type="wicri:Area/Pmc/Corpus">000693</idno><idno type="wicri:Area/Pmc/Curation">000692</idno><idno type="wicri:Area/Pmc/Checkpoint">000134</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Immunolocalization of cyclotides in plant cells, tissues and organ supports their role in host defense</title><author><name sortKey="Slazak, Blazej" sort="Slazak, Blazej" uniqKey="Slazak B" first="Blazej" last="Slazak">Blazej Slazak</name><affiliation wicri:level="1"><nlm:aff id="Aff1">W. Szafer Institute of Botany, Polish Academy of Science, 46 Lubicz St, 31-512 Cracow, Poland</nlm:aff><country xml:lang="fr">Pologne</country><wicri:regionArea>W. Szafer Institute of Botany, Polish Academy of Science, 46 Lubicz St, 31-512 Cracow</wicri:regionArea><wicri:noRegion>31-512 Cracow</wicri:noRegion></affiliation><affiliation wicri:level="4"><nlm:aff id="Aff2">Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Center, Box 574, 751 23 Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Center, Box 574, 751 23 Uppsala</wicri:regionArea><orgName type="university">Université d'Uppsala</orgName><placeName><settlement type="city">Uppsala</settlement><region nuts="1">Svealand</region><region nuts="1">East Middle Sweden</region></placeName></affiliation></author><author><name sortKey="Kapusta, Malgorzata" sort="Kapusta, Malgorzata" uniqKey="Kapusta M" first="Małgorzata" last="Kapusta">Małgorzata Kapusta</name><affiliation wicri:level="1"><nlm:aff id="Aff3">Department of Plant Cytology and Embryology, Faculty of Biology, University of Gdańsk, 59 Wita Stwosza St, 80-308 Gdańsk, Poland</nlm:aff><country xml:lang="fr">Pologne</country><wicri:regionArea>Department of Plant Cytology and Embryology, Faculty of Biology, University of Gdańsk, 59 Wita Stwosza St, 80-308 Gdańsk</wicri:regionArea><wicri:noRegion>80-308 Gdańsk</wicri:noRegion></affiliation></author><author><name sortKey="Malik, Sohaib" sort="Malik, Sohaib" uniqKey="Malik S" first="Sohaib" last="Malik">Sohaib Malik</name><affiliation wicri:level="4"><nlm:aff id="Aff2">Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Center, Box 574, 751 23 Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Center, Box 574, 751 23 Uppsala</wicri:regionArea><orgName type="university">Université d'Uppsala</orgName><placeName><settlement type="city">Uppsala</settlement><region nuts="1">Svealand</region><region nuts="1">East Middle Sweden</region></placeName></affiliation></author><author><name sortKey="Bohdanowicz, Jerzy" sort="Bohdanowicz, Jerzy" uniqKey="Bohdanowicz J" first="Jerzy" last="Bohdanowicz">Jerzy Bohdanowicz</name><affiliation wicri:level="1"><nlm:aff id="Aff3">Department of Plant Cytology and Embryology, Faculty of Biology, University of Gdańsk, 59 Wita Stwosza St, 80-308 Gdańsk, Poland</nlm:aff><country xml:lang="fr">Pologne</country><wicri:regionArea>Department of Plant Cytology and Embryology, Faculty of Biology, University of Gdańsk, 59 Wita Stwosza St, 80-308 Gdańsk</wicri:regionArea><wicri:noRegion>80-308 Gdańsk</wicri:noRegion></affiliation></author><author><name sortKey="Kuta, El Bieta" sort="Kuta, El Bieta" uniqKey="Kuta E" first="El Bieta" last="Kuta">El Bieta Kuta</name><affiliation wicri:level="1"><nlm:aff id="Aff4">Department of Plant Cytology and Embryology, Institute of Botany, Jagiellonian University, 9 Gronostajowa St, 30-387 Cracow, Poland</nlm:aff><country xml:lang="fr">Pologne</country><wicri:regionArea>Department of Plant Cytology and Embryology, Institute of Botany, Jagiellonian University, 9 Gronostajowa St, 30-387 Cracow</wicri:regionArea><wicri:noRegion>30-387 Cracow</wicri:noRegion></affiliation></author><author><name sortKey="Malec, Przemyslaw" sort="Malec, Przemyslaw" uniqKey="Malec P" first="Przemysław" last="Malec">Przemysław Malec</name><affiliation wicri:level="1"><nlm:aff id="Aff5">Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 7 Gronostajowa St, 30-387 Cracow, Poland</nlm:aff><country xml:lang="fr">Pologne</country><wicri:regionArea>Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 7 Gronostajowa St, 30-387 Cracow</wicri:regionArea><wicri:noRegion>30-387 Cracow</wicri:noRegion></affiliation></author><author><name sortKey="Goransson, Ulf" sort="Goransson, Ulf" uniqKey="Goransson U" first="Ulf" last="Göransson">Ulf Göransson</name><affiliation wicri:level="4"><nlm:aff id="Aff2">Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Center, Box 574, 751 23 Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Center, Box 574, 751 23 Uppsala</wicri:regionArea><orgName type="university">Université d'Uppsala</orgName><placeName><settlement type="city">Uppsala</settlement><region nuts="1">Svealand</region><region nuts="1">East Middle Sweden</region></placeName></affiliation></author></analytic><series><title level="j">Planta</title><idno type="ISSN">0032-0935</idno><idno type="eISSN">1432-2048</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><title><bold><italic>Main conclusion</italic></bold></title><p><bold>The distribution of cyclotides was visualized in plant cells, tissues and organs using immunohistochemistry. Finding of cyclotides in tissues potentially vulnerable to pathogen attacks supports their role as defense molecules.</bold></p><p>The cyclotide family of plant peptides is characterized by the cyclic cystine knot motif and its diverse biological activities. Given their insecticidal and antimicrobial properties, the role of cyclotides <italic>in planta</italic> is probably associated with host defense. Our current understanding of the cellular compartmentalization of cyclotides in the vacuole is based on indirect studies on transgenic model plants that do not express cyclotides naturally. Matrix-assisted laser desorption ionization (MALDI) imaging has also been used to study the distribution of cyclotides, but the technique’s resolution was insufficient to determine their tissue or cell distribution. To avoid the limitations of these approaches, immunohistochemical visualization methods were used. Antibodies were raised in rabbits using cycloviolacin O2 (cyO2), and their specificity was determined by Western and dot blot experiments. Slides for immunohistochemical analysis were prepared from leaf, petiole and root fragments of <italic>Viola odorata</italic> and <italic>Viola uliginosa</italic>, and specimens were visualized using indirect epifluorescence microscopy. The antibodies against cyclotides were specific against selected bracelet cyclotides with high similarity (cyO2, cyO3, cyO8, cyO13) and suitable for immunohistochemistry. The tissue distribution of the cyclotides visualized in this way is consistent with their proposed role in host defense—relatively large quantities were observed in the leaf and petiole epidermis in both <italic>Viola</italic> species. Cyclotides were also found in vascular tissue in all the assessed plant organs. The vacuole storage of cyclotides was directly shown.</p><sec><title>Electronic supplementary material</title><p>The online version of this article (doi:10.1007/s00425-016-2562-y) contains supplementary material, which is available to authorized users.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Aichler, M" uniqKey="Aichler M">M Aichler</name></author><author><name sortKey="Walch, A" uniqKey="Walch A">A Walch</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Atkins, C" uniqKey="Atkins C">C Atkins</name></author><author><name sortKey="Smith, Pmc" uniqKey="Smith P">PMC Smith</name></author><author><name sortKey="Rodriguez Medina, C" uniqKey="Rodriguez Medina C">C Rodriguez-Medina</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Burman, R" uniqKey="Burman R">R Burman</name></author><author><name sortKey="Gunasekera, S" uniqKey="Gunasekera S">S 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Planta</journal-id><journal-id journal-id-type="iso-abbrev">Planta</journal-id><journal-title-group><journal-title>Planta</journal-title></journal-title-group><issn pub-type="ppub">0032-0935</issn><issn pub-type="epub">1432-2048</issn><publisher><publisher-name>Springer Berlin Heidelberg</publisher-name><publisher-loc>Berlin/Heidelberg</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">27394154</article-id><article-id pub-id-type="pmc">5052299</article-id><article-id pub-id-type="publisher-id">2562</article-id><article-id pub-id-type="doi">10.1007/s00425-016-2562-y</article-id><article-categories><subj-group subj-group-type="heading"><subject>Original Article</subject></subj-group></article-categories><title-group><article-title>Immunolocalization of cyclotides in plant cells, tissues and organ supports their role in host defense</article-title></title-group><contrib-group><contrib contrib-type="author" corresp="yes"><name><surname>Slazak</surname><given-names>Blazej</given-names></name><address><phone>+48 660 450 799</phone><email>b.slazak@botany.pl</email></address><xref ref-type="aff" rid="Aff1">1</xref><xref ref-type="aff" rid="Aff2">2</xref></contrib><contrib contrib-type="author"><name><surname>Kapusta</surname><given-names>Małgorzata</given-names></name><xref ref-type="aff" rid="Aff3">3</xref></contrib><contrib contrib-type="author"><name><surname>Malik</surname><given-names>Sohaib</given-names></name><xref ref-type="aff" rid="Aff2">2</xref></contrib><contrib contrib-type="author"><name><surname>Bohdanowicz</surname><given-names>Jerzy</given-names></name><xref ref-type="aff" rid="Aff3">3</xref></contrib><contrib contrib-type="author"><name><surname>Kuta</surname><given-names>Elżbieta</given-names></name><xref ref-type="aff" rid="Aff4">4</xref></contrib><contrib contrib-type="author"><name><surname>Malec</surname><given-names>Przemysław</given-names></name><xref ref-type="aff" rid="Aff5">5</xref></contrib><contrib contrib-type="author"><name><surname>Göransson</surname><given-names>Ulf</given-names></name><xref ref-type="aff" rid="Aff2">2</xref></contrib><aff id="Aff1"><label>1</label>W. Szafer Institute of Botany, Polish Academy of Science, 46 Lubicz St, 31-512 Cracow, Poland</aff><aff id="Aff2"><label>2</label>Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Center, Box 574, 751 23 Uppsala, Sweden</aff><aff id="Aff3"><label>3</label>Department of Plant Cytology and Embryology, Faculty of Biology, University of Gdańsk, 59 Wita Stwosza St, 80-308 Gdańsk, Poland</aff><aff id="Aff4"><label>4</label>Department of Plant Cytology and Embryology, Institute of Botany, Jagiellonian University, 9 Gronostajowa St, 30-387 Cracow, Poland</aff><aff id="Aff5"><label>5</label>Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 7 Gronostajowa St, 30-387 Cracow, Poland</aff></contrib-group><pub-date pub-type="epub"><day>9</day><month>7</month><year>2016</year></pub-date><pub-date pub-type="pmc-release"><day>9</day><month>7</month><year>2016</year></pub-date><pub-date pub-type="ppub"><year>2016</year></pub-date><volume>244</volume><issue>5</issue><fpage>1029</fpage><lpage>1040</lpage><history><date date-type="received"><day>12</day><month>2</month><year>2016</year></date><date date-type="accepted"><day>20</day><month>6</month><year>2016</year></date></history><permissions><copyright-statement>© The Author(s) 2016</copyright-statement><license license-type="OpenAccess"><license-p><bold>Open Access</bold>This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link>), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.</license-p></license></permissions><abstract id="Abs1"><title><bold><italic>Main conclusion</italic></bold></title><p><bold>The distribution of cyclotides was visualized in plant cells, tissues and organs using immunohistochemistry. Finding of cyclotides in tissues potentially vulnerable to pathogen attacks supports their role as defense molecules.</bold></p><p>The cyclotide family of plant peptides is characterized by the cyclic cystine knot motif and its diverse biological activities. Given their insecticidal and antimicrobial properties, the role of cyclotides <italic>in planta</italic> is probably associated with host defense. Our current understanding of the cellular compartmentalization of cyclotides in the vacuole is based on indirect studies on transgenic model plants that do not express cyclotides naturally. Matrix-assisted laser desorption ionization (MALDI) imaging has also been used to study the distribution of cyclotides, but the technique’s resolution was insufficient to determine their tissue or cell distribution. To avoid the limitations of these approaches, immunohistochemical visualization methods were used. Antibodies were raised in rabbits using cycloviolacin O2 (cyO2), and their specificity was determined by Western and dot blot experiments. Slides for immunohistochemical analysis were prepared from leaf, petiole and root fragments of <italic>Viola odorata</italic> and <italic>Viola uliginosa</italic>, and specimens were visualized using indirect epifluorescence microscopy. The antibodies against cyclotides were specific against selected bracelet cyclotides with high similarity (cyO2, cyO3, cyO8, cyO13) and suitable for immunohistochemistry. The tissue distribution of the cyclotides visualized in this way is consistent with their proposed role in host defense—relatively large quantities were observed in the leaf and petiole epidermis in both <italic>Viola</italic> species. Cyclotides were also found in vascular tissue in all the assessed plant organs. The vacuole storage of cyclotides was directly shown.</p><sec><title>Electronic supplementary material</title><p>The online version of this article (doi:10.1007/s00425-016-2562-y) contains supplementary material, which is available to authorized users.</p></sec></abstract><kwd-group xml:lang="en"><title>Keywords</title><kwd>Cyclotides</kwd><kwd>Immunohistochemistry</kwd><kwd>Host defense peptides</kwd><kwd><italic>Viola</italic></kwd></kwd-group><funding-group><award-group><funding-source><institution-wrap><institution-id institution-id-type="FundRef">http://dx.doi.org/10.13039/501100001862</institution-id><institution>Svenska Forskningsrådet Formas</institution></institution-wrap></funding-source><award-id>621-2007-5167</award-id><principal-award-recipient><name><surname>Göransson</surname><given-names>Ulf</given-names></name></principal-award-recipient></award-group><award-group><funding-source><institution>Stiftelsen för Strategisk Forskning (SE)</institution></funding-source><award-id>F06-0058</award-id><principal-award-recipient><name><surname>Göransson</surname><given-names>Ulf</given-names></name></principal-award-recipient></award-group><award-group><funding-source><institution>Polish Botany Foundation</institution></funding-source></award-group><award-group><funding-source><institution-wrap><institution-id institution-id-type="FundRef">http://dx.doi.org/10.13039/501100004382</institution-id><institution>Polska Akademia Nauk</institution></institution-wrap></funding-source></award-group><award-group><funding-source><institution>Ahlquists Stiftelse (Ahlquist Foundation)</institution></funding-source></award-group></funding-group><custom-meta-group><custom-meta><meta-name>issue-copyright-statement</meta-name><meta-value>© Springer-Verlag Berlin Heidelberg 2016</meta-value></custom-meta></custom-meta-group></article-meta></front></pmc><affiliations><list><country><li>Pologne</li><li>Suède</li></country><region><li>East Middle Sweden</li><li>Svealand</li></region><settlement><li>Uppsala</li></settlement><orgName><li>Université d'Uppsala</li></orgName></list><tree><country name="Pologne"><noRegion><name sortKey="Slazak, Blazej" sort="Slazak, Blazej" uniqKey="Slazak B" first="Blazej" last="Slazak">Blazej Slazak</name></noRegion><name sortKey="Bohdanowicz, Jerzy" sort="Bohdanowicz, Jerzy" uniqKey="Bohdanowicz J" first="Jerzy" last="Bohdanowicz">Jerzy Bohdanowicz</name><name sortKey="Kapusta, Malgorzata" sort="Kapusta, Malgorzata" uniqKey="Kapusta M" first="Małgorzata" last="Kapusta">Małgorzata Kapusta</name><name sortKey="Kuta, El Bieta" sort="Kuta, El Bieta" uniqKey="Kuta E" first="El Bieta" last="Kuta">El Bieta Kuta</name><name sortKey="Malec, Przemyslaw" sort="Malec, Przemyslaw" uniqKey="Malec P" first="Przemysław" last="Malec">Przemysław Malec</name></country><country name="Suède"><region name="Svealand"><name sortKey="Slazak, Blazej" sort="Slazak, Blazej" uniqKey="Slazak B" first="Blazej" last="Slazak">Blazej Slazak</name></region><name sortKey="Goransson, Ulf" sort="Goransson, Ulf" uniqKey="Goransson U" first="Ulf" last="Göransson">Ulf Göransson</name><name sortKey="Malik, Sohaib" sort="Malik, Sohaib" uniqKey="Malik S" first="Sohaib" last="Malik">Sohaib Malik</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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