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Hyper-accumulation of starch and oil in a Chlamydomonas mutant affected in a plant-specific DYRK kinase

Identifieur interne : 001D40 ( Main/Exploration ); précédent : 001D39; suivant : 001D41

Hyper-accumulation of starch and oil in a Chlamydomonas mutant affected in a plant-specific DYRK kinase

Auteurs : Miriam Schulz-Raffelt [France, Allemagne] ; Vincent Chochois [France] ; Pascaline Auroy [France] ; Stéphan Cuiné [France] ; Emmanuelle Billon [France] ; David Dauvillée [France] ; Yonghua Li-Beisson [France] ; Gilles Peltier [France]

Source :

RBID : PMC:4782384

Abstract

Background

Because of their high biomass productivity and their ability to accumulate high levels of energy-rich reserve compounds such as oils or starch, microalgae represent a promising feedstock for the production of biofuel. Accumulation of reserve compounds takes place when microalgae face adverse situations such as nutrient shortage, conditions which also provoke a stop in cell division, and down-regulation of photosynthesis. Despite growing interest in microalgal biofuels, little is known about molecular mechanisms controlling carbon reserve formation. In order to discover new regulatory mechanisms, and identify genes of interest to boost the potential of microalgae for biofuel production, we developed a forward genetic approach in the model microalga Chlamydomonas reinhardtii.

Results

By screening an insertional mutant library on the ability of mutants to accumulate and re-mobilize reserve compounds, we isolated a Chlamydomonas mutant (starch degradation 1, std1) deficient for a dual-specificity tyrosine-phosphorylation-regulated kinase (DYRK). The std1 mutant accumulates higher levels of starch and oil than wild-type and maintains a higher photosynthetic activity under nitrogen starvation. Phylogenetic analysis revealed that this kinase (named DYRKP) belongs to a plant-specific subgroup of the evolutionarily conserved DYRK kinase family. Furthermore, hyper-accumulation of storage compounds occurs in std1 mostly under low light in photoautotrophic condition, suggesting that the kinase normally acts under conditions of low energy status to limit reserve accumulation.

Conclusions

The DYRKP kinase is proposed to act as a negative regulator of the sink capacity of photosynthetic cells that integrates nutrient and energy signals. Inactivation of the kinase strongly boosts accumulation of reserve compounds under photoautotrophic nitrogen deprivation and allows maintaining high photosynthetic activity. The DYRKP kinase therefore represents an attractive target for improving the energy density of microalgae or crop plants.

Electronic supplementary material

The online version of this article (doi:10.1186/s13068-016-0469-2) contains supplementary material, which is available to authorized users.


Url:
DOI: 10.1186/s13068-016-0469-2
PubMed: 26958078
PubMed Central: 4782384


Affiliations:


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<nlm:aff id="Aff3">Aix Marseille Université, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13284 Marseille, France</nlm:aff>
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<settlement type="city">Marseille</settlement>
</placeName>
<orgName type="university">Université d'Aix-Marseille</orgName>
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<nlm:aff id="Aff5">Molecular Biotechnology and Systems Biology, TU Kaiserslautern, Paul-Ehrlich-Straße 23, 67663 Kaiserslautern, Germany</nlm:aff>
<country xml:lang="fr">Allemagne</country>
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</affiliation>
</author>
<author>
<name sortKey="Chochois, Vincent" sort="Chochois, Vincent" uniqKey="Chochois V" first="Vincent" last="Chochois">Vincent Chochois</name>
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<country xml:lang="fr">France</country>
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<orgName type="university">Université d'Aix-Marseille</orgName>
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<settlement type="city">Marseille</settlement>
<region type="region" nuts="2">Provence-Alpes-Côte d'Azur</region>
</placeName>
</affiliation>
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<nlm:aff id="Aff2">CNRS, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13108 Saint-Paul-lez-Durance, France</nlm:aff>
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<wicri:noRegion>13108 Saint-Paul-lez-Durance</wicri:noRegion>
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<settlement type="city">Marseille</settlement>
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<settlement type="city">Marseille</settlement>
</placeName>
<orgName type="university">Université d'Aix-Marseille</orgName>
</affiliation>
<affiliation>
<nlm:aff id="Aff6">Research School of Biology College of Medicine, Biology and Environment, Linneaus Building 134, The Australian National University, Canberra, ACT 2601 Australia</nlm:aff>
<wicri:noCountry code="subfield">ACT 2601 Australia</wicri:noCountry>
</affiliation>
</author>
<author>
<name sortKey="Auroy, Pascaline" sort="Auroy, Pascaline" uniqKey="Auroy P" first="Pascaline" last="Auroy">Pascaline Auroy</name>
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<nlm:aff id="Aff1">CEA, CNRS, Aix-Marseille Université, Institut de Biosciences et Biotechnologies Aix Marseille, Laboratoire de Bioénergétique et Biotechnologie des Bactéries et Microalgues, CEA Cadarache, 13108 Saint-Paul-lez-Durance, France</nlm:aff>
<country xml:lang="fr">France</country>
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<orgName type="university">Université d'Aix-Marseille</orgName>
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<settlement type="city">Marseille</settlement>
<region type="region" nuts="2">Provence-Alpes-Côte d'Azur</region>
</placeName>
</affiliation>
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<nlm:aff id="Aff2">CNRS, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13108 Saint-Paul-lez-Durance, France</nlm:aff>
<country xml:lang="fr">France</country>
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<wicri:noRegion>13108 Saint-Paul-lez-Durance</wicri:noRegion>
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<affiliation wicri:level="4">
<nlm:aff id="Aff3">Aix Marseille Université, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13284 Marseille, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>Aix Marseille Université, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13284 Marseille</wicri:regionArea>
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<settlement type="city">Marseille</settlement>
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<settlement type="city">Marseille</settlement>
</placeName>
<orgName type="university">Université d'Aix-Marseille</orgName>
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</author>
<author>
<name sortKey="Cuine, Stephan" sort="Cuine, Stephan" uniqKey="Cuine S" first="Stéphan" last="Cuiné">Stéphan Cuiné</name>
<affiliation wicri:level="4">
<nlm:aff id="Aff1">CEA, CNRS, Aix-Marseille Université, Institut de Biosciences et Biotechnologies Aix Marseille, Laboratoire de Bioénergétique et Biotechnologie des Bactéries et Microalgues, CEA Cadarache, 13108 Saint-Paul-lez-Durance, France</nlm:aff>
<country xml:lang="fr">France</country>
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<wicri:noRegion>13108 Saint-Paul-lez-Durance</wicri:noRegion>
<orgName type="university">Université d'Aix-Marseille</orgName>
<placeName>
<settlement type="city">Marseille</settlement>
<region type="region" nuts="2">Provence-Alpes-Côte d'Azur</region>
</placeName>
</affiliation>
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<nlm:aff id="Aff2">CNRS, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13108 Saint-Paul-lez-Durance, France</nlm:aff>
<country xml:lang="fr">France</country>
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<wicri:noRegion>13108 Saint-Paul-lez-Durance</wicri:noRegion>
</affiliation>
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<nlm:aff id="Aff3">Aix Marseille Université, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13284 Marseille, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>Aix Marseille Université, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13284 Marseille</wicri:regionArea>
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<placeName>
<settlement type="city">Marseille</settlement>
<region type="région" nuts="2">Provence-Alpes-Côte d'Azur</region>
<settlement type="city">Marseille</settlement>
</placeName>
<orgName type="university">Université d'Aix-Marseille</orgName>
</affiliation>
</author>
<author>
<name sortKey="Billon, Emmanuelle" sort="Billon, Emmanuelle" uniqKey="Billon E" first="Emmanuelle" last="Billon">Emmanuelle Billon</name>
<affiliation wicri:level="4">
<nlm:aff id="Aff1">CEA, CNRS, Aix-Marseille Université, Institut de Biosciences et Biotechnologies Aix Marseille, Laboratoire de Bioénergétique et Biotechnologie des Bactéries et Microalgues, CEA Cadarache, 13108 Saint-Paul-lez-Durance, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>CEA, CNRS, Aix-Marseille Université, Institut de Biosciences et Biotechnologies Aix Marseille, Laboratoire de Bioénergétique et Biotechnologie des Bactéries et Microalgues, CEA Cadarache, 13108 Saint-Paul-lez-Durance</wicri:regionArea>
<wicri:noRegion>13108 Saint-Paul-lez-Durance</wicri:noRegion>
<orgName type="university">Université d'Aix-Marseille</orgName>
<placeName>
<settlement type="city">Marseille</settlement>
<region type="region" nuts="2">Provence-Alpes-Côte d'Azur</region>
</placeName>
</affiliation>
<affiliation wicri:level="1">
<nlm:aff id="Aff2">CNRS, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13108 Saint-Paul-lez-Durance, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>CNRS, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13108 Saint-Paul-lez-Durance</wicri:regionArea>
<wicri:noRegion>13108 Saint-Paul-lez-Durance</wicri:noRegion>
<wicri:noRegion>13108 Saint-Paul-lez-Durance</wicri:noRegion>
</affiliation>
<affiliation wicri:level="4">
<nlm:aff id="Aff3">Aix Marseille Université, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13284 Marseille, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>Aix Marseille Université, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13284 Marseille</wicri:regionArea>
<wicri:noRegion>13284 Marseille</wicri:noRegion>
<placeName>
<settlement type="city">Marseille</settlement>
<region type="région" nuts="2">Provence-Alpes-Côte d'Azur</region>
<settlement type="city">Marseille</settlement>
</placeName>
<orgName type="university">Université d'Aix-Marseille</orgName>
</affiliation>
</author>
<author>
<name sortKey="Dauvillee, David" sort="Dauvillee, David" uniqKey="Dauvillee D" first="David" last="Dauvillée">David Dauvillée</name>
<affiliation wicri:level="1">
<nlm:aff id="Aff4">UMR8576, CNRS, Université des Sciences et Technologies de Lille, 59655 Villeneuve d’Ascq, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>UMR8576, CNRS, Université des Sciences et Technologies de Lille, 59655 Villeneuve d’Ascq</wicri:regionArea>
<wicri:noRegion>59655 Villeneuve d’Ascq</wicri:noRegion>
<wicri:noRegion>59655 Villeneuve d’Ascq</wicri:noRegion>
</affiliation>
</author>
<author>
<name sortKey="Li Beisson, Yonghua" sort="Li Beisson, Yonghua" uniqKey="Li Beisson Y" first="Yonghua" last="Li-Beisson">Yonghua Li-Beisson</name>
<affiliation wicri:level="4">
<nlm:aff id="Aff1">CEA, CNRS, Aix-Marseille Université, Institut de Biosciences et Biotechnologies Aix Marseille, Laboratoire de Bioénergétique et Biotechnologie des Bactéries et Microalgues, CEA Cadarache, 13108 Saint-Paul-lez-Durance, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>CEA, CNRS, Aix-Marseille Université, Institut de Biosciences et Biotechnologies Aix Marseille, Laboratoire de Bioénergétique et Biotechnologie des Bactéries et Microalgues, CEA Cadarache, 13108 Saint-Paul-lez-Durance</wicri:regionArea>
<wicri:noRegion>13108 Saint-Paul-lez-Durance</wicri:noRegion>
<orgName type="university">Université d'Aix-Marseille</orgName>
<placeName>
<settlement type="city">Marseille</settlement>
<region type="region" nuts="2">Provence-Alpes-Côte d'Azur</region>
</placeName>
</affiliation>
<affiliation wicri:level="1">
<nlm:aff id="Aff2">CNRS, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13108 Saint-Paul-lez-Durance, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>CNRS, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13108 Saint-Paul-lez-Durance</wicri:regionArea>
<wicri:noRegion>13108 Saint-Paul-lez-Durance</wicri:noRegion>
<wicri:noRegion>13108 Saint-Paul-lez-Durance</wicri:noRegion>
</affiliation>
<affiliation wicri:level="4">
<nlm:aff id="Aff3">Aix Marseille Université, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13284 Marseille, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>Aix Marseille Université, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13284 Marseille</wicri:regionArea>
<wicri:noRegion>13284 Marseille</wicri:noRegion>
<placeName>
<settlement type="city">Marseille</settlement>
<region type="région" nuts="2">Provence-Alpes-Côte d'Azur</region>
<settlement type="city">Marseille</settlement>
</placeName>
<orgName type="university">Université d'Aix-Marseille</orgName>
</affiliation>
</author>
<author>
<name sortKey="Peltier, Gilles" sort="Peltier, Gilles" uniqKey="Peltier G" first="Gilles" last="Peltier">Gilles Peltier</name>
<affiliation wicri:level="4">
<nlm:aff id="Aff1">CEA, CNRS, Aix-Marseille Université, Institut de Biosciences et Biotechnologies Aix Marseille, Laboratoire de Bioénergétique et Biotechnologie des Bactéries et Microalgues, CEA Cadarache, 13108 Saint-Paul-lez-Durance, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>CEA, CNRS, Aix-Marseille Université, Institut de Biosciences et Biotechnologies Aix Marseille, Laboratoire de Bioénergétique et Biotechnologie des Bactéries et Microalgues, CEA Cadarache, 13108 Saint-Paul-lez-Durance</wicri:regionArea>
<wicri:noRegion>13108 Saint-Paul-lez-Durance</wicri:noRegion>
<orgName type="university">Université d'Aix-Marseille</orgName>
<placeName>
<settlement type="city">Marseille</settlement>
<region type="region" nuts="2">Provence-Alpes-Côte d'Azur</region>
</placeName>
</affiliation>
<affiliation wicri:level="1">
<nlm:aff id="Aff2">CNRS, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13108 Saint-Paul-lez-Durance, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>CNRS, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13108 Saint-Paul-lez-Durance</wicri:regionArea>
<wicri:noRegion>13108 Saint-Paul-lez-Durance</wicri:noRegion>
<wicri:noRegion>13108 Saint-Paul-lez-Durance</wicri:noRegion>
</affiliation>
<affiliation wicri:level="4">
<nlm:aff id="Aff3">Aix Marseille Université, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13284 Marseille, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>Aix Marseille Université, Biologie Végétale et Microbiologie Environnementale, UMR7265, 13284 Marseille</wicri:regionArea>
<wicri:noRegion>13284 Marseille</wicri:noRegion>
<placeName>
<settlement type="city">Marseille</settlement>
<region type="région" nuts="2">Provence-Alpes-Côte d'Azur</region>
<settlement type="city">Marseille</settlement>
</placeName>
<orgName type="university">Université d'Aix-Marseille</orgName>
</affiliation>
</author>
</analytic>
<series>
<title level="j">Biotechnology for Biofuels</title>
<idno type="eISSN">1754-6834</idno>
<imprint>
<date when="2016">2016</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc>
<textClass></textClass>
</profileDesc>
</teiHeader>
<front>
<div type="abstract" xml:lang="en">
<sec>
<title>Background</title>
<p>Because of their high biomass productivity and their ability to accumulate high levels of energy-rich reserve compounds such as oils or starch, microalgae represent a promising feedstock for the production of biofuel. Accumulation of reserve compounds takes place when microalgae face adverse situations such as nutrient shortage, conditions which also provoke a stop in cell division, and down-regulation of photosynthesis. Despite growing interest in microalgal biofuels, little is known about molecular mechanisms controlling carbon reserve formation. In order to discover new regulatory mechanisms, and identify genes of interest to boost the potential of microalgae for biofuel production, we developed a forward genetic approach in the model microalga
<italic>Chlamydomonas reinhardtii</italic>
.</p>
</sec>
<sec>
<title>Results</title>
<p>By screening an insertional mutant library on the ability of mutants to accumulate and re-mobilize reserve compounds, we isolated a
<italic>Chlamydomonas</italic>
mutant (
<italic>starch degradation 1, std1</italic>
) deficient for a dual-specificity tyrosine-phosphorylation-regulated kinase (DYRK). The
<italic>std1</italic>
mutant accumulates higher levels of starch and oil than wild-type and maintains a higher photosynthetic activity under nitrogen starvation. Phylogenetic analysis revealed that this kinase (named DYRKP) belongs to a plant-specific subgroup of the evolutionarily conserved DYRK kinase family. Furthermore, hyper-accumulation of storage compounds occurs in
<italic>std1</italic>
mostly under low light in photoautotrophic condition, suggesting that the kinase normally acts under conditions of low energy status to limit reserve accumulation.</p>
</sec>
<sec>
<title>Conclusions</title>
<p>The DYRKP kinase is proposed to act as a negative regulator of the sink capacity of photosynthetic cells that integrates nutrient and energy signals. Inactivation of the kinase strongly boosts accumulation of reserve compounds under photoautotrophic nitrogen deprivation and allows maintaining high photosynthetic activity. The DYRKP kinase therefore represents an attractive target for improving the energy density of microalgae or crop plants.</p>
</sec>
<sec>
<title>Electronic supplementary material</title>
<p>The online version of this article (doi:10.1186/s13068-016-0469-2) contains supplementary material, which is available to authorized users.</p>
</sec>
</div>
</front>
<back>
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<name sortKey="Stecher, G" uniqKey="Stecher G">G Stecher</name>
</author>
<author>
<name sortKey="Nei, M" uniqKey="Nei M">M Nei</name>
</author>
<author>
<name sortKey="Kumar, S" uniqKey="Kumar S">S Kumar</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Liu, Cm" uniqKey="Liu C">CM Liu</name>
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<author>
<name sortKey="Willmund, F" uniqKey="Willmund F">F Willmund</name>
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<author>
<name sortKey="Whitelegge, Jp" uniqKey="Whitelegge J">JP Whitelegge</name>
</author>
<author>
<name sortKey="Hawat, S" uniqKey="Hawat S">S Hawat</name>
</author>
<author>
<name sortKey="Knapp, B" uniqKey="Knapp B">B Knapp</name>
</author>
<author>
<name sortKey="Lodha, M" uniqKey="Lodha M">M Lodha</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Li Beisson, Y" uniqKey="Li Beisson Y">Y Li-Beisson</name>
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<author>
<name sortKey="Shorrosh, B" uniqKey="Shorrosh B">B Shorrosh</name>
</author>
<author>
<name sortKey="Beisson, F" uniqKey="Beisson F">F Beisson</name>
</author>
<author>
<name sortKey="Andersson, Mx" uniqKey="Andersson M">MX Andersson</name>
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<author>
<name sortKey="Arondel, V" uniqKey="Arondel V">V Arondel</name>
</author>
<author>
<name sortKey="Bates, Pd" uniqKey="Bates P">PD Bates</name>
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</analytic>
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<biblStruct>
<analytic>
<author>
<name sortKey="Schreiber, U" uniqKey="Schreiber U">U Schreiber</name>
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<name sortKey="Schliwa, U" uniqKey="Schliwa U">U Schliwa</name>
</author>
<author>
<name sortKey="Bilger, W" uniqKey="Bilger W">W Bilger</name>
</author>
</analytic>
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<country>
<li>Allemagne</li>
<li>France</li>
</country>
<region>
<li>Provence-Alpes-Côte d'Azur</li>
</region>
<settlement>
<li>Marseille</li>
<li>Saint-Paul-lez-Durance</li>
</settlement>
<orgName>
<li>Université d'Aix-Marseille</li>
</orgName>
</list>
<tree>
<country name="France">
<region name="Provence-Alpes-Côte d'Azur">
<name sortKey="Schulz Raffelt, Miriam" sort="Schulz Raffelt, Miriam" uniqKey="Schulz Raffelt M" first="Miriam" last="Schulz-Raffelt">Miriam Schulz-Raffelt</name>
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<name sortKey="Auroy, Pascaline" sort="Auroy, Pascaline" uniqKey="Auroy P" first="Pascaline" last="Auroy">Pascaline Auroy</name>
<name sortKey="Auroy, Pascaline" sort="Auroy, Pascaline" uniqKey="Auroy P" first="Pascaline" last="Auroy">Pascaline Auroy</name>
<name sortKey="Auroy, Pascaline" sort="Auroy, Pascaline" uniqKey="Auroy P" first="Pascaline" last="Auroy">Pascaline Auroy</name>
<name sortKey="Billon, Emmanuelle" sort="Billon, Emmanuelle" uniqKey="Billon E" first="Emmanuelle" last="Billon">Emmanuelle Billon</name>
<name sortKey="Billon, Emmanuelle" sort="Billon, Emmanuelle" uniqKey="Billon E" first="Emmanuelle" last="Billon">Emmanuelle Billon</name>
<name sortKey="Billon, Emmanuelle" sort="Billon, Emmanuelle" uniqKey="Billon E" first="Emmanuelle" last="Billon">Emmanuelle Billon</name>
<name sortKey="Chochois, Vincent" sort="Chochois, Vincent" uniqKey="Chochois V" first="Vincent" last="Chochois">Vincent Chochois</name>
<name sortKey="Chochois, Vincent" sort="Chochois, Vincent" uniqKey="Chochois V" first="Vincent" last="Chochois">Vincent Chochois</name>
<name sortKey="Chochois, Vincent" sort="Chochois, Vincent" uniqKey="Chochois V" first="Vincent" last="Chochois">Vincent Chochois</name>
<name sortKey="Cuine, Stephan" sort="Cuine, Stephan" uniqKey="Cuine S" first="Stéphan" last="Cuiné">Stéphan Cuiné</name>
<name sortKey="Cuine, Stephan" sort="Cuine, Stephan" uniqKey="Cuine S" first="Stéphan" last="Cuiné">Stéphan Cuiné</name>
<name sortKey="Cuine, Stephan" sort="Cuine, Stephan" uniqKey="Cuine S" first="Stéphan" last="Cuiné">Stéphan Cuiné</name>
<name sortKey="Dauvillee, David" sort="Dauvillee, David" uniqKey="Dauvillee D" first="David" last="Dauvillée">David Dauvillée</name>
<name sortKey="Li Beisson, Yonghua" sort="Li Beisson, Yonghua" uniqKey="Li Beisson Y" first="Yonghua" last="Li-Beisson">Yonghua Li-Beisson</name>
<name sortKey="Li Beisson, Yonghua" sort="Li Beisson, Yonghua" uniqKey="Li Beisson Y" first="Yonghua" last="Li-Beisson">Yonghua Li-Beisson</name>
<name sortKey="Li Beisson, Yonghua" sort="Li Beisson, Yonghua" uniqKey="Li Beisson Y" first="Yonghua" last="Li-Beisson">Yonghua Li-Beisson</name>
<name sortKey="Peltier, Gilles" sort="Peltier, Gilles" uniqKey="Peltier G" first="Gilles" last="Peltier">Gilles Peltier</name>
<name sortKey="Peltier, Gilles" sort="Peltier, Gilles" uniqKey="Peltier G" first="Gilles" last="Peltier">Gilles Peltier</name>
<name sortKey="Peltier, Gilles" sort="Peltier, Gilles" uniqKey="Peltier G" first="Gilles" last="Peltier">Gilles Peltier</name>
<name sortKey="Schulz Raffelt, Miriam" sort="Schulz Raffelt, Miriam" uniqKey="Schulz Raffelt M" first="Miriam" last="Schulz-Raffelt">Miriam Schulz-Raffelt</name>
<name sortKey="Schulz Raffelt, Miriam" sort="Schulz Raffelt, Miriam" uniqKey="Schulz Raffelt M" first="Miriam" last="Schulz-Raffelt">Miriam Schulz-Raffelt</name>
</country>
<country name="Allemagne">
<noRegion>
<name sortKey="Schulz Raffelt, Miriam" sort="Schulz Raffelt, Miriam" uniqKey="Schulz Raffelt M" first="Miriam" last="Schulz-Raffelt">Miriam Schulz-Raffelt</name>
</noRegion>
</country>
</tree>
</affiliations>
</record>

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