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<document id="07E841C0FCFA30404EA13281547D35D2" ID-DOI="10.1016/j.phytochem.2014.09.021" ID-ISSN="1873-3700" ID-Zenodo-Dep="10490604" IM.bibliography_approvedBy="felipe" IM.illustrations_approvedBy="diego" IM.materialsCitations_approvedBy="diego" IM.metadata_approvedBy="felipe" IM.taxonomicNames_approvedBy="diego" IM.treatments_approvedBy="diego" checkinTime="1704963693688" checkinUser="felipe" docAuthor="Tian, Yuan, Amand, Séverine, Buisson, Didier, Kunz, Caroline, Hachette, François, Dupont, Joëlle, Nay, Bastien &amp; Prado, Soizic" docDate="2014" docId="0388C53DFF942E76FFC9FF276A5FFE8F" docLanguage="en" docName="Phytochemistry.108.95-101.pdf" docOrigin="Phytochemistry 108" docSource="http://dx.doi.org/10.1016/j.phytochem.2014.09.021" docStyle="DocumentStyle:9E596C34F4E94307D29315B03ACE1007.6:Phytochemistry.2014-2019.journal_article" docStyleId="9E596C34F4E94307D29315B03ACE1007" docStyleName="Phytochemistry.2014-2019.journal_article" docStyleVersion="6" docTitle="Cephalotaxus harringtonia subsp. crude" docType="treatment" docVersion="1" lastPageNumber="97" masterDocId="FFB1BD45FF952E75FF9EF8676825FFDA" masterDocTitle="The fungal leaf endophyte Paraconiothyrium variabile specifically metabolizes the host-plant metabolome for its own benefit" masterLastPageNumber="101" masterPageNumber="95" pageNumber="96" updateTime="1705416565815" updateUser="diego">
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<mods:title id="9F3AF2BAB37F0C6ADD26C91B4B741E57">The fungal leaf endophyte Paraconiothyrium variabile specifically metabolizes the host-plant metabolome for its own benefit</mods:title>
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<mods:namePart id="07282F496500110F892648B391967E00">Tian, Yuan</mods:namePart>
<mods:affiliation id="7B36BE4B3D5F9048AE1342D84F8F8334">Molécules de Communication et Adaptation des Microorganismes (MCAM), UMR 7245 CNRS / Muséum National d’Histoire Naturelle, Paris, France</mods:affiliation>
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<mods:namePart id="1E2D85A8051E7895F67CE74E7FFF74FC">Amand, Séverine</mods:namePart>
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<mods:namePart id="906BAC731F862E1B0EC3728518F64660">Buisson, Didier</mods:namePart>
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<mods:namePart id="54698026D23537E80B05FD1AFDDB2F23">Kunz, Caroline</mods:namePart>
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<mods:name id="32BBADDAEC0CDEB5D9C8F1405935D3CE" type="personal">
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<mods:namePart id="8761CBDDB20D9F0ABB5265765D94099C">Hachette, François</mods:namePart>
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<mods:namePart id="A39741488ADA1A8501291D5A32420458">Dupont, Joëlle</mods:namePart>
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<mods:namePart id="D3B29E8DBFEEC053C5E38468CA0928AD">Nay, Bastien</mods:namePart>
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<mods:namePart id="570A1137A5CABB5A12284FADD8CD3659">Prado, Soizic</mods:namePart>
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<mods:title id="76FB561E54BB8664C65E7E2A92F65418">Phytochemistry</mods:title>
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<mods:date id="FCBB116AFE6F18B81499B214F642B318">2014</mods:date>
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<mods:identifier id="188B261F19350E68F39EFEDC3A7BD541" type="DOI">10.1016/j.phytochem.2014.09.021</mods:identifier>
<mods:identifier id="790E0D7BE93025EDD9AC53D201BCEEED" type="ISSN">1873-3700</mods:identifier>
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<treatment id="0388C53DFF942E76FFC9FF276A5FFE8F" LSID="urn:lsid:plazi:treatment:0388C53DFF942E76FFC9FF276A5FFE8F" httpUri="http://treatment.plazi.org/id/0388C53DFF942E76FFC9FF276A5FFE8F" lastPageId="3" lastPageNumber="97" pageId="1" pageNumber="96">
<subSubSection id="C33B27A0FF942E74FFC9FF2769CBF8AA" pageId="1" pageNumber="96" type="nomenclature">
<paragraph id="8B9E742BFF942E74FFC9FF2769CBF8AA" blockId="1.[87,695,1856,1904]" pageId="1" pageNumber="96">
<heading id="D0D6C347FF942E74FFC9FF2769CBF8AA" fontSize="8" level="3" pageId="1" pageNumber="96" reason="8">
<emphasis id="B955A839FF942E74FFC9FF2769CBF8AA" italics="true" pageId="1" pageNumber="96">
2.1. Metabolomics analysis of the
<taxonomicName id="4C210FA8FF942E74FE01FF276A15F88E" box="[415,560,1856,1876]" class="Pinopsida" family="Cephalotaxaceae" genus="Cephalotaxus" kingdom="Plantae" order="Pinales" pageId="1" pageNumber="96" phylum="Tracheophyta" rank="subSpecies" species="harringtonia" subSpecies="crude">C. harringtonia</taxonomicName>
crude extract transformed by the endophyte
<taxonomicName id="4C210FA8FF942E74FE1DFF3B69CBF8AA" box="[387,494,1884,1904]" class="Dothideomycetes" family="Didymosphaeriaceae" genus="Paraconiothyrium" kingdom="Fungi" order="Pleosporales" pageId="1" pageNumber="96" phylum="Ascomycota" rank="species" species="variabile">P. variabile</taxonomicName>
</emphasis>
</heading>
</paragraph>
</subSubSection>
<subSubSection id="C33B27A0FF942E76FFE8FFF26A5FFE8F" lastPageId="3" lastPageNumber="98" pageId="1" pageNumber="96" type="description">
<paragraph id="8B9E742BFF942E74FFE8FFF26C71FEC7" blockId="1.[87,757,1940,1988]" lastBlockId="1.[805,1475,182,592]" pageId="1" pageNumber="96">
Biotransformations of the crude extract of
<taxonomicName id="4C210FA8FF942E74FDBFFFF36A95F872" box="[545,688,1940,1960]" class="Pinopsida" family="Cephalotaxaceae" genus="Cephalotaxus" kingdom="Plantae" order="Pinales" pageId="1" pageNumber="96" phylum="Tracheophyta" rank="species" species="harringtonia">
<emphasis id="B955A839FF942E74FDBFFFF36A95F872" box="[545,688,1940,1960]" italics="true" pageId="1" pageNumber="96">C. harringtonia</emphasis>
</taxonomicName>
by the endophyte
<taxonomicName id="4C210FA8FF942E74FF50FFD7691AF81E" box="[206,319,1968,1988]" class="Dothideomycetes" family="Didymosphaeriaceae" genus="Paraconiothyrium" kingdom="Fungi" order="Pleosporales" pageId="1" pageNumber="96" phylum="Ascomycota" rank="species" species="variabile">
<emphasis id="B955A839FF942E74FF50FFD7691AF81E" box="[206,319,1968,1988]" italics="true" pageId="1" pageNumber="96">P. variabile</emphasis>
</taxonomicName>
were monitored by standardized LC–
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fingerprints at 16, 20, 40, and 60 h post inoculation (hpi). Using the
<collectionCode id="ED30ECEEFF942E74FCCFF8B56BA8FF3F" box="[849,909,210,229]" pageId="1" pageNumber="96">XCMS</collectionCode>
software, the protonated ions of the biotransformed metabolites at the different hpi were compared to the control crude extract sample (
<emphasis id="B955A839FF942E74FB97F96E6C34FEC7" box="[1033,1041,265,285]" italics="true" pageId="1" pageNumber="96">t</emphasis>
= 0 h).
</paragraph>
<paragraph id="8B9E742BFF942E74FCDAF9416D20FD8A" blockId="1.[805,1475,182,592]" pageId="1" pageNumber="96">
From these analyses more than one hundred protonated ions showed a decreasing intensity in the course of the biotransformation. To analyze the most abundant and relevant ions, we selected only those with an intensity superior to 10,000 and a relative fold change, as defined by
<collectionCode id="ED30ECEEFF942E74FB94F9F16C62FE73" box="[1034,1095,406,425]" pageId="1" pageNumber="96">XCMS</collectionCode>
, superior to 100. With these criteria, five metabolites appeared to be transformed predominantly by the fungus. Indeed, the pseudomolecular ions [
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+H]
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at
<emphasis id="B955A839FF942E74FA01F9AB6DE7FE3A" box="[1439,1474,460,480]" italics="true" pageId="1" pageNumber="96">m/z</emphasis>
371.21, 373.22, 579.14, 621.18 and 651.19 underwent a significant decrease of intensity 40 h after incubation with the mycelium of
<taxonomicName id="4C210FA8FF942E74FA2FFA636B5CFDEE" class="Dothideomycetes" family="Didymosphaeriaceae" genus="Paraconiothyrium" kingdom="Fungi" order="Pleosporales" pageId="1" pageNumber="96" phylum="Ascomycota" rank="species" species="variabile">
<emphasis id="B955A839FF942E74FA2FFA636B5CFDEE" italics="true" pageId="1" pageNumber="96">P. variabile</emphasis>
</taxonomicName>
(
<figureCitation id="131A68AEFF942E74FC14FA466BE6FDEE" box="[906,963,545,564]" captionStart="Fig" captionStartId="2.[113,139,742,756]" captionTargetBox="[272,1334,181,712]" captionTargetId="figure-190@2.[272,1334,181,712]" captionTargetPageId="2" captionText="Fig. 1. Decrease of the relative intensity of the protonated ions [M+H] + at m/z 651.19, 621.18, 579.14, 373.22 and 371.21 of the metabolites mainly biotransformed by P. variabile in the crude extract of C. harringtonia at 16, 20, 40 and 60 h post-inoculation by comparison to control (t = 0) generated by XCMS software. Stars indicate significant differences calculated by XCMS by comparison to control (t = 0) at t = 40 h and t = 60 h. ⁄⁄p &lt;0.01 (n = 3)." figureDoi="http://doi.org/10.5281/zenodo.10490606" httpUri="https://zenodo.org/record/10490606/files/figure.png" pageId="1" pageNumber="96">Fig. 1</figureCitation>
). Among these metabolites, those at
<emphasis id="B955A839FF942E74FAD5FA476D4AFDEE" box="[1355,1391,544,564]" italics="true" pageId="1" pageNumber="96">m/z</emphasis>
579.14, 621.18 and 651.19 were the most transformed.
</paragraph>
<paragraph id="8B9E742BFF942E74FCBBFAF66C0FFD1B" blockId="1.[805,1370,657,705]" pageId="1" pageNumber="96">
<emphasis id="B955A839FF942E74FCBBFAF66C0FFD1B" italics="true" pageId="1" pageNumber="96">
2.2. Isolation and characterization of the main compounds
<heading id="D0D6C347FF942E74FCBBFACA6C0FFD1B" box="[805,1066,685,705]" fontSize="8" level="3" pageId="1" pageNumber="96" reason="8">
metabolized by
<taxonomicName id="4C210FA8FF942E74FC20FACA6C0FFD1B" box="[958,1066,685,705]" class="Dothideomycetes" family="Didymosphaeriaceae" genus="Paraconiothyrium" kingdom="Fungi" order="Pleosporales" pageId="1" pageNumber="96" phylum="Ascomycota" rank="species" species="variabile">P. variabile</taxonomicName>
</heading>
</emphasis>
</paragraph>
<paragraph id="8B9E742BFF942E74FCDAFA816CECFB92" blockId="1.[805,1475,741,1988]" pageId="1" pageNumber="96">
We started by analyzing the compounds that had been most transformed generating ions at
<emphasis id="B955A839FF942E74FBEBFB676CBCFCCE" box="[1141,1177,768,788]" italics="true" pageId="1" pageNumber="96">m/z</emphasis>
579.14, 621.18 and 651.19. We isolated them from the crude extract by semi-preparative
<collectionCode id="ED30ECEEFF942E74FA13FB7A6DE7FCEA" box="[1421,1474,797,816]" pageId="1" pageNumber="96">HPLC</collectionCode>
guided by
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detection, giving the tree main metabolites at
<emphasis id="B955A839FF942E74FA01FB5F6DE7FC96" box="[1439,1474,824,844]" italics="true" pageId="1" pageNumber="96">m/z</emphasis>
579.14, 621.18 and 651.19. The analysis of 1D and 2D NMR spectra, along with the
<collectionCode id="ED30ECEEFF942E74FC4CFB166BD7FC5E" box="[978,1010,881,900]" country="Italy" lsid="urn:lsid:biocol.org:col:14396" name="Herbarium Messanaensis, Università di Messina" pageId="1" pageNumber="96" type="Herbarium">MS</collectionCode>
/
<collectionCode id="ED30ECEEFF942E74FC67FB166C3CFC5E" box="[1017,1049,881,900]" country="Italy" lsid="urn:lsid:biocol.org:col:14396" name="Herbarium Messanaensis, Università di Messina" pageId="1" pageNumber="96" type="Herbarium">MS</collectionCode>
fragmentations and bibliographic data, allowed unambiguous characterization of these compounds. The collision-induced dissociation (CID) experiments on compound (
<emphasis id="B955A839FF942E74FCB3FBA26B1FFC02" bold="true" box="[813,826,965,984]" pageId="1" pageNumber="96">1</emphasis>
), which displays a pseudomolecular protonated ion [
<collectionCode id="ED30ECEEFF942E74FAFCFBA26D5DFC02" box="[1378,1400,965,984]" country="Germany" lsid="urn:lsid:biocol.org:col:15637" name="Botanische Staatssammlung München" pageId="1" pageNumber="96" type="Herbarium">M</collectionCode>
+H]
<superScript id="7C54D963FF942E74FA03FBA66D83FC17" attach="left" box="[1437,1446,961,973]" fontSize="5" pageId="1" pageNumber="96">+</superScript>
at
<emphasis id="B955A839FF942E74FCBBFB876B6DFC2E" box="[805,840,992,1012]" italics="true" pageId="1" pageNumber="96">m/z</emphasis>
: 579.14, led to two main fragments at
<emphasis id="B955A839FF942E74FB4CFB876CD3FC2E" box="[1234,1270,992,1012]" italics="true" pageId="1" pageNumber="96">m/z</emphasis>
433 and 271, corresponding to the successive loss of 146 u and 162 u. These were attributed to the loss of a deoxyhexose and a hexose suggesting that compound
<emphasis id="B955A839FF942E74FC59FC536BF1FB9D" bold="true" box="[967,980,1076,1095]" pageId="1" pageNumber="96">1</emphasis>
is a diglycoside (
<figureCitation id="131A68AEFF942E74FB1BFC536C99FB92" box="[1157,1212,1076,1096]" captionStart="Fig" captionStartId="2.[113,139,1581,1595]" captionTargetBox="[348,1265,861,1501]" captionTargetPageId="2" captionText="Fig. 2. Structures of the main compounds (1–3) isolated from the leaves of C. harringtonia (A) biotransformed by the endophytic fungus P. variabile and their fragments at m/z provided by fragmentation of their respective protonated ion [M+H] + at m/z 579.14, 621.18, 651.19 from CID experiments. Key NMR correlations (HMBC) are also represented for localization of the acetate group of compound (2) and are identical for compound (3)." figureDoi="http://doi.org/10.5281/zenodo.10490608" httpUri="https://zenodo.org/record/10490608/files/figure.png" pageId="1" pageNumber="96">Fig. 2</figureCitation>
).
</paragraph>
<paragraph id="8B9E742BFF942E74FCDAFC376D07F913" blockId="1.[805,1475,741,1988]" pageId="1" pageNumber="96">
The
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H MNR spectrum of compound
<quantity id="4CD9D9CEFF942E74FB5DFC376CC9FBB9" box="[1219,1260,1104,1123]" metricMagnitude="-2" metricUnit="m" metricValue="2.54" pageId="1" pageNumber="96" unit="in" value="1.0">
<emphasis id="B955A839FF942E74FB5DFC376CF5FBB9" bold="true" box="[1219,1232,1104,1123]" pageId="1" pageNumber="96">1</emphasis>
in
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CD
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OD displayed an olefinic signal at
<subScript id="17A5766EFF942E74FC45FC0B6BD7FB5B" attach="left" box="[987,1010,1132,1153]" fontSize="5" pageId="1" pageNumber="96">dH</subScript>
6.51 (1H, s, H-3) characteristic of a flavone structure. This compound appeared to be dihydroxylated at position 4
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and 7, as indicated by signals at
<subScript id="17A5766EFF942E74FB5CFCC36CFCFB63" attach="left" box="[1218,1241,1188,1209]" fontSize="5" pageId="1" pageNumber="96">dH</subScript>
7.81 (2H, d,
<emphasis id="B955A839FF942E74FAFFFCC46D4DFB6D" box="[1377,1384,1187,1207]" italics="true" pageId="1" pageNumber="96">J</emphasis>
= 8.7 Hz, H-2
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</emphasis>
, H-6
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</emphasis>
),
<subScript id="17A5766EFF942E74FC04FCA76B94FB0F" attach="left" box="[922,945,1216,1237]" fontSize="5" pageId="1" pageNumber="96">dH</subScript>
6.92 (2H, d,
<emphasis id="B955A839FF942E74FBAAFCD86C1EFB09" box="[1076,1083,1215,1235]" italics="true" pageId="1" pageNumber="96">J</emphasis>
= 8.7 Hz, H-3
<emphasis id="B955A839FF942E74FB5CFCDA6CE3FB10" box="[1218,1222,1213,1226]" italics="true" pageId="1" pageNumber="96">
<superScript id="7C54D963FF942E74FB5CFCDA6CE3FB10" attach="none" box="[1218,1222,1213,1226]" fontSize="5" pageId="1" pageNumber="96">0</superScript>
</emphasis>
, H-5
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<superScript id="7C54D963FF942E74FB67FCDA6CD8FB10" attach="none" box="[1273,1277,1213,1226]" fontSize="5" pageId="1" pageNumber="96">0</superScript>
</emphasis>
) which are characteristic of a
<emphasis id="B955A839FF942E74FC02FCBC6B8DFB35" box="[924,936,1243,1263]" italics="true" pageId="1" pageNumber="96">p</emphasis>
-hydroxyphenyl ring, and at
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6.63 (1H, d,
<emphasis id="B955A839FF942E74FAFFFCBC6D4DFB35" box="[1377,1384,1243,1263]" italics="true" pageId="1" pageNumber="96">J</emphasis>
= 2.2 Hz, H-8) and
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6.60 (1H, d,
<emphasis id="B955A839FF942E74FB83FC916C01FAD0" box="[1053,1060,1270,1290]" italics="true" pageId="1" pageNumber="96">J</emphasis>
= 2.2 Hz, H-6) which reveal the presence of a phloroglucinol ring. Two anomeric protons at
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5.37 (1H, d,
<emphasis id="B955A839FF942E74FCBBFD496B09FA98" box="[805,812,1326,1346]" italics="true" pageId="1" pageNumber="96">J</emphasis>
= 1.5 Hz, H-1
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<superScript id="7C54D963FF942E74FC2FFD4B6B99FAE3" attach="none" box="[945,956,1324,1337]" fontSize="5" pageId="1" pageNumber="96">000</superScript>
</emphasis>
) and
<subScript id="17A5766EFF942E74FC6BFD486C29FA9F" attach="left" box="[1013,1036,1327,1349]" fontSize="5" pageId="1" pageNumber="96">dH</subScript>
5.30 were also (1H, d,
<emphasis id="B955A839FF942E74FB72FD496CD6FA98" box="[1260,1267,1326,1346]" italics="true" pageId="1" pageNumber="96">J</emphasis>
= 6.6 Hz, H-1
<emphasis id="B955A839FF942E74FAE6FD4B6DA5FAE3" box="[1400,1408,1324,1337]" italics="true" pageId="1" pageNumber="96">
<superScript id="7C54D963FF942E74FAE6FD4B6DA5FAE3" attach="none" box="[1400,1408,1324,1337]" fontSize="5" pageId="1" pageNumber="96">00</superScript>
</emphasis>
) present, characteristic of the diglycoside part, and a methyl proton at
<subScript id="17A5766EFF942E74FCBBFD006B1EFAA6" attach="left" box="[805,827,1383,1404]" fontSize="5" pageId="1" pageNumber="96">dH</subScript>
1.12 (3H, d,
<emphasis id="B955A839FF942E74FC5DFD016BEFFAA0" box="[963,970,1382,1402]" italics="true" pageId="1" pageNumber="96">J</emphasis>
= 7.0 Hz, H-6
<emphasis id="B955A839FF942E74FBCCFD036C78FAAB" box="[1106,1117,1380,1393]" italics="true" pageId="1" pageNumber="96">
<superScript id="7C54D963FF942E74FBCCFD036C78FAAB" attach="left" box="[1106,1117,1380,1393]" fontSize="5" pageId="1" pageNumber="96">000</superScript>
</emphasis>
) showing the deoxyhexose nature of one of the sugar unit. The interglycosidic linkage as well as the attachment position of the diglycoside to the aglycone was confirmed by rigorous analysis of the
<superScript id="7C54D963FF942E74FB27FDD06CE7FA19" attach="right" box="[1209,1218,1463,1475]" fontSize="5" pageId="1" pageNumber="96">1</superScript>
H
<emphasis id="B955A839FF942E74FB4CFDDA6CC1FA14" bold="true" box="[1234,1252,1469,1486]" pageId="1" pageNumber="96">
<collectionCode id="ED30ECEEFF942E74FB4CFDDA6CC1FA14" box="[1234,1252,1469,1486]" country="USA" lsid="urn:lsid:biocol.org:col:15406" name="Harvard University - Arnold Arboretum" pageId="1" pageNumber="96" type="Herbarium">A</collectionCode>
</emphasis>
<superScript id="7C54D963FF942E74FB7AFDD06CC8FA19" attach="both" box="[1252,1261,1463,1475]" fontSize="5" pageId="1" pageNumber="96">1</superScript>
H COSY and
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H
<emphasis id="B955A839FF942E74FA0FFDDA6D86FA14" bold="true" box="[1425,1443,1469,1486]" pageId="1" pageNumber="96">
<collectionCode id="ED30ECEEFF942E74FA0FFDDA6D86FA14" box="[1425,1443,1469,1486]" country="USA" lsid="urn:lsid:biocol.org:col:15406" name="Harvard University - Arnold Arboretum" pageId="1" pageNumber="96" type="Herbarium">A</collectionCode>
</emphasis>
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<collectionCode id="ED30ECEEFF942E74FA2BFDDC6DE7FA14" box="[1461,1474,1467,1486]" country="Denmark" name="University of Copenhagen" pageId="1" pageNumber="96" type="Herbarium">C</collectionCode>
HMBC correlations, both showing 2-bond and 3-bond scalar couplings. The configuration of the glycosidic linkage of the glucopyranoside and rhamnopyranoside were determined to be
<emphasis id="B955A839FF942E74FAF0FE686D5FF9F8" bold="true" box="[1390,1402,1551,1570]" italics="true" pageId="1" pageNumber="96">β</emphasis>
and
<emphasis id="B955A839FF942E74FA2EFE6B6D98F9F8" bold="true" box="[1456,1469,1548,1570]" italics="true" pageId="1" pageNumber="96">oi</emphasis>
, respectively, on the basis of the
<superScript id="7C54D963FF942E74FB3AFE416C88F9E8" attach="right" box="[1188,1197,1574,1586]" fontSize="5" pageId="1" pageNumber="96">3</superScript>
<emphasis id="B955A839FF942E74FB33FE4E6C91F9E7" box="[1197,1204,1577,1597]" italics="true" pageId="1" pageNumber="96">J</emphasis>
<subScript id="17A5766EFF942E74FB2AFE546CF8F9E3" attach="left" box="[1204,1245,1585,1599]" fontSize="3" pageId="1" pageNumber="96">
1
<emphasis id="B955A839FF942E74FB23FE566CE6F9E3" box="[1213,1219,1585,1593]" italics="true" pageId="1" pageNumber="96">00</emphasis>
–2
<emphasis id="B955A839FF942E74FB49FE566CF8F9E3" box="[1239,1245,1585,1593]" italics="true" pageId="1" pageNumber="96">00</emphasis>
</subScript>
(6.6 Hz) and
<superScript id="7C54D963FF942E74FA1DFE416DA9F9E8" attach="right" box="[1411,1420,1574,1586]" fontSize="5" pageId="1" pageNumber="96">3</superScript>
<emphasis id="B955A839FF942E74FA12FE4E6DB6F9E7" box="[1420,1427,1577,1597]" italics="true" pageId="1" pageNumber="96">J</emphasis>
<subScript id="17A5766EFF942E74FA0DFE546DE5F9E3" attach="left" box="[1427,1472,1585,1599]" fontSize="3" pageId="1" pageNumber="96">
1
<emphasis id="B955A839FF942E74FA02FE566D81F9E3" box="[1436,1444,1585,1593]" italics="true" pageId="1" pageNumber="96">000</emphasis>
–2
<emphasis id="B955A839FF942E74FA26FE566DE5F9E3" box="[1464,1472,1585,1593]" italics="true" pageId="1" pageNumber="96">000</emphasis>
</subScript>
(1.5 Hz) values of the anomeric protons. Thus, from these results and from comparison with bibliographic data, the structure of this compound was assigned as apigenin 5-
<emphasis id="B955A839FF942E74FB56FE1A6CF2F94B" box="[1224,1239,1661,1681]" italics="true" pageId="1" pageNumber="96">
<collectionCode id="ED30ECEEFF942E74FB56FE1A6CF2F94B" box="[1224,1239,1661,1681]" country="Norway" lsid="urn:lsid:biocol.org:col:13093" name="Botanical Museum - University of Oslo" pageId="1" pageNumber="96" type="Herbarium">O</collectionCode>
</emphasis>
-a- L- rhamnopyranosyl- (1
<emphasis id="B955A839FF942E74FCA7FEFC6B6AF977" box="[825,847,1691,1709]" italics="true" pageId="1" pageNumber="96">+</emphasis>
3)-
<emphasis id="B955A839FF942E74FCF3FEFD6B5CF977" bold="true" box="[877,889,1690,1709]" italics="true" pageId="1" pageNumber="96">β</emphasis>
-D- glucopyranoside (
<emphasis id="B955A839FF942E74FBCCFEFD6C7AF977" bold="true" box="[1106,1119,1690,1709]" pageId="1" pageNumber="96">1</emphasis>
) which was previously isolated from a
<taxonomicName id="4C210FA8FF942E74FCF1FED26BCAF913" authorityName="Siebold &amp; Zuccarini ex Endlicher" authorityYear="1842" box="[879,1007,1717,1737]" class="Pinopsida" family="Cephalotaxaceae" genus="Cephalotaxus" kingdom="Plantae" order="Pinales" pageId="1" pageNumber="96" phylum="Tracheophyta" rank="genus">
<emphasis id="B955A839FF942E74FCF1FED26BCAF913" box="[879,1007,1717,1737]" italics="true" pageId="1" pageNumber="96">Cephalotaxus</emphasis>
</taxonomicName>
tree (
<bibRefCitation id="EFB009DAFF942E74FBB2FED16CE2F913" author="Bae, K. &amp; Jin, W. &amp; Thuong, P. T. &amp; Min, B. S. &amp; Na, M. &amp; Lee, Y. M. &amp; Kang, S. S." box="[1068,1223,1718,1737]" pageId="1" pageNumber="96" pagination="409 - 413" refId="ref6749" refString="Bae, K., Jin, W., Thuong, P. T., Min, B. S., Na, M., Lee, Y. M., Kang, S. S., 2007. A new flavonoid glycoside from the leaf of Cephalotaxus koreana. Fitoterapia 78, 409 - 413." type="journal article" year="2007">Bae et al., 2007</bibRefCitation>
) (
<figureCitation id="131A68AEFF942E74FB40FED16D30F913" box="[1246,1301,1718,1737]" captionStart="Fig" captionStartId="2.[113,139,1581,1595]" captionTargetBox="[348,1265,861,1501]" captionTargetPageId="2" captionText="Fig. 2. Structures of the main compounds (1–3) isolated from the leaves of C. harringtonia (A) biotransformed by the endophytic fungus P. variabile and their fragments at m/z provided by fragmentation of their respective protonated ion [M+H] + at m/z 579.14, 621.18, 651.19 from CID experiments. Key NMR correlations (HMBC) are also represented for localization of the acetate group of compound (2) and are identical for compound (3)." figureDoi="http://doi.org/10.5281/zenodo.10490608" httpUri="https://zenodo.org/record/10490608/files/figure.png" pageId="1" pageNumber="96">Fig. 2</figureCitation>
).
</paragraph>
<paragraph id="8B9E742BFF942E74FCDAFEB56C42F81E" blockId="1.[805,1475,741,1988]" pageId="1" pageNumber="96">
In the same way, we identified by one-and two-dimensional NMR as well as
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/
<collectionCode id="ED30ECEEFF942E74FC75FE896C2EF8DB" box="[1003,1035,1774,1793]" country="Italy" lsid="urn:lsid:biocol.org:col:14396" name="Herbarium Messanaensis, Università di Messina" pageId="1" pageNumber="96" type="Herbarium">MS</collectionCode>
experiments, compound
<emphasis id="B955A839FF942E74FA95FE896D3DF8DB" bold="true" box="[1291,1304,1774,1793]" pageId="1" pageNumber="96">2</emphasis>
which displayed a pseudomolecular protonated ion [
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+H]
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at
<emphasis id="B955A839FF942E74FB73FF6E6D35F8C7" box="[1261,1296,1801,1821]" italics="true" pageId="1" pageNumber="96">m/z</emphasis>
: 621.18 as apigenin 5-
<emphasis id="B955A839FF942E74FCEAFF436BA6F8E2" box="[884,899,1828,1848]" italics="true" pageId="1" pageNumber="96">
<collectionCode id="ED30ECEEFF942E74FCEAFF436BA6F8E2" box="[884,899,1828,1848]" country="Norway" lsid="urn:lsid:biocol.org:col:13093" name="Botanical Museum - University of Oslo" pageId="1" pageNumber="96" type="Herbarium">O</collectionCode>
</emphasis>
-a- L- rhamnopyranosyl-(1
<emphasis id="B955A839FF942E74FB1DFF416CBCF8E2" box="[1155,1177,1830,1848]" italics="true" pageId="1" pageNumber="96">+</emphasis>
2)-(6
<emphasis id="B955A839FF942E74FB55FF456CC9F8E2" box="[1227,1260,1826,1849]" italics="true" pageId="1" pageNumber="96">
<superScript id="7C54D963FF942E74FB55FF456CF6F8F5" attach="none" box="[1227,1235,1826,1839]" fontSize="5" pageId="1" pageNumber="96">00</superScript>
-
<collectionCode id="ED30ECEEFF942E74FB43FF436CC9F8E2" box="[1245,1260,1828,1848]" country="Norway" lsid="urn:lsid:biocol.org:col:13093" name="Botanical Museum - University of Oslo" pageId="1" pageNumber="96" type="Herbarium">O</collectionCode>
</emphasis>
-acetyl)-
<emphasis id="B955A839FF942E74FADFFF416D68F8E3" bold="true" box="[1345,1357,1830,1849]" italics="true" pageId="1" pageNumber="96">β</emphasis>
-D- glucopyranoside, also known as euryanoside (
<emphasis id="B955A839FF942E74FB56FF266CF0F88E" bold="true" box="[1224,1237,1857,1876]" pageId="1" pageNumber="96">2</emphasis>
) (
<bibRefCitation id="EFB009DAFF942E74FB6CFF266D90F88F" author="Inada, A. &amp; Fujiwara, M. &amp; Kakimoto, L. &amp; Kitamura, F. &amp; Toya, H. &amp; Konishi, M. &amp; Nakanishi, T. &amp; Murata, H." box="[1266,1461,1857,1877]" pageId="1" pageNumber="96" pagination="2819 - 2821" refId="ref7137" refString="Inada, A., Fujiwara, M., Kakimoto, L., Kitamura, F., Toya, H., Konishi, M., Nakanishi, T., Murata, H., 1989. Structure of a new acylated flavonoid glycoside, euryanoside, from flowers of Eurya-Japonica Thunb. Chem. Pharm. Bull. 37, 2819 - 2821." type="journal article" year="1989">Inada et al., 1989</bibRefCitation>
). Compound
<emphasis id="B955A839FF942E74FC07FF3A6B83F8AA" bold="true" box="[921,934,1885,1904]" pageId="1" pageNumber="96">3</emphasis>
with an
<emphasis id="B955A839FF942E74FB9FFF3B6C00F8AA" box="[1025,1061,1884,1904]" italics="true" pageId="1" pageNumber="96">m/z</emphasis>
651.19 was assigned as chrysoeriol 5-
<emphasis id="B955A839FF942E74FA34FF3B6D9CF8AA" box="[1450,1465,1884,1904]" italics="true" pageId="1" pageNumber="96">
<collectionCode id="ED30ECEEFF942E74FA34FF3B6D9CF8AA" box="[1450,1465,1884,1904]" country="Norway" lsid="urn:lsid:biocol.org:col:13093" name="Botanical Museum - University of Oslo" pageId="1" pageNumber="96" type="Herbarium">O</collectionCode>
</emphasis>
- a- L- rhamnopyranosyl-(1
<emphasis id="B955A839FF942E74FBBDFF1D6C1CF856" box="[1059,1081,1914,1932]" italics="true" pageId="1" pageNumber="96">+</emphasis>
4)-(6
<emphasis id="B955A839FF942E74FBF0FF116CAAF856" box="[1134,1167,1910,1933]" italics="true" pageId="1" pageNumber="96">
<superScript id="7C54D963FF942E74FBF0FF116C53F859" attach="none" box="[1134,1142,1910,1923]" fontSize="5" pageId="1" pageNumber="96">00</superScript>
-
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</emphasis>
-acetyl)-
<emphasis id="B955A839FF942E74FB76FF1E6CD1F856" bold="true" box="[1256,1268,1913,1932]" italics="true" pageId="1" pageNumber="96">β</emphasis>
-D- glucopyranoside, another flavone glycoside previously isolated from a
<taxonomicName id="4C210FA8FF942E74FADDFFF36DE7F872" authorityName="Siebold &amp; Zuccarini ex Endlicher" authorityYear="1842" box="[1347,1474,1940,1960]" class="Pinopsida" family="Cephalotaxaceae" genus="Cephalotaxus" kingdom="Plantae" order="Pinales" pageId="1" pageNumber="96" phylum="Tracheophyta" rank="genus">
<emphasis id="B955A839FF942E74FADDFFF36DE7F872" box="[1347,1474,1940,1960]" italics="true" pageId="1" pageNumber="96">Cephalotaxus</emphasis>
</taxonomicName>
tree (
<bibRefCitation id="EFB009DAFF942E74FCC5FFD66C2BF81E" author="Zhang, Y. Q. &amp; Xu, Z. H. &amp; Deng, Y. L." box="[859,1038,1969,1988]" pageId="1" pageNumber="96" pagination="1386 - 1391" refId="ref8589" refString="Zhang, Y. Q., Xu, Z. H., Deng, Y. L., 2011. Characterization of flavone glycosides and aglycones in Cephalotaxus sinensis by HPLC - DAD - MS. Anal. Methods 3, 1386 - 1391." type="journal article" year="2011">Zhang et al., 2011</bibRefCitation>
) (
<figureCitation id="131A68AEFF942E74FBBAFFD66C7FF81E" box="[1060,1114,1969,1988]" captionStart="Fig" captionStartId="2.[113,139,1581,1595]" captionTargetBox="[348,1265,861,1501]" captionTargetPageId="2" captionText="Fig. 2. Structures of the main compounds (1–3) isolated from the leaves of C. harringtonia (A) biotransformed by the endophytic fungus P. variabile and their fragments at m/z provided by fragmentation of their respective protonated ion [M+H] + at m/z 579.14, 621.18, 651.19 from CID experiments. Key NMR correlations (HMBC) are also represented for localization of the acetate group of compound (2) and are identical for compound (3)." figureDoi="http://doi.org/10.5281/zenodo.10490608" httpUri="https://zenodo.org/record/10490608/files/figure.png" pageId="1" pageNumber="96">Fig. 2</figureCitation>
).
</paragraph>
<caption id="DF5E24A3FF972E77FFEFFA816BE3FCF8" ID-DOI="http://doi.org/10.5281/zenodo.10490606" ID-Zenodo-Dep="10490606" httpUri="https://zenodo.org/record/10490606/files/figure.png" pageId="2" pageNumber="97" startId="2.[113,139,742,756]" targetBox="[272,1334,181,712]" targetPageId="2" targetType="figure">
<paragraph id="8B9E742BFF972E77FFEFFA816BE3FCF8" blockId="2.[113,1502,738,802]" pageId="2" pageNumber="97">
<emphasis id="B955A839FF972E77FFEFFA816881FD2E" bold="true" box="[113,164,742,756]" pageId="2" pageNumber="97">Fig. 1.</emphasis>
Decrease of the relative intensity of the protonated ions [M+H]
<superScript id="7C54D963FF972E77FD2CFA856A9CFD31" attach="left" box="[690,697,738,747]" fontSize="4" pageId="2" pageNumber="97">+</superScript>
at
<emphasis id="B955A839FF972E77FD4AFA826AD5FD2E" box="[724,752,741,756]" italics="true" pageId="2" pageNumber="97">m/z</emphasis>
651.19, 621.18, 579.14, 373.22 and 371.21 of the metabolites mainly biotransformed by
<taxonomicName id="4C210FA8FF972E77FA51FA826891FCD1" class="Dothideomycetes" family="Didymosphaeriaceae" genus="Paraconiothyrium" kingdom="Fungi" order="Pleosporales" pageId="2" pageNumber="97" phylum="Ascomycota" rank="species" species="variabile">
<emphasis id="B955A839FF972E77FA51FA826891FCD1" italics="true" pageId="2" pageNumber="97">P. variabile</emphasis>
</taxonomicName>
in the crude extract of
<taxonomicName id="4C210FA8FF972E77FEEEFA9C69C6FCD1" box="[368,483,763,779]" class="Pinopsida" family="Cephalotaxaceae" genus="Cephalotaxus" kingdom="Plantae" order="Pinales" pageId="2" pageNumber="97" phylum="Tracheophyta" rank="species" species="harringtonia">
<emphasis id="B955A839FF972E77FEEEFA9C69C6FCD1" box="[368,483,763,779]" italics="true" pageId="2" pageNumber="97">C. harringtonia</emphasis>
</taxonomicName>
at 16, 20, 40 and 60 h post-inoculation by comparison to control (
<emphasis id="B955A839FF972E77FC66FA9B6BDBFCD1" box="[1016,1022,764,779]" italics="true" pageId="2" pageNumber="97">t</emphasis>
= 0) generated by XCMS software. Stars indicate significant differences calculated by XCMS by comparison to control (
<emphasis id="B955A839FF972E77FDD1FB746A70FCF8" box="[591,597,787,802]" italics="true" pageId="2" pageNumber="97">t</emphasis>
= 0) at
<emphasis id="B955A839FF972E77FD0DFB746ABCFCF8" box="[659,665,787,802]" italics="true" pageId="2" pageNumber="97">t</emphasis>
= 40 h and
<emphasis id="B955A839FF972E77FD69FB746AD8FCF8" box="[759,765,787,802]" italics="true" pageId="2" pageNumber="97">t</emphasis>
= 60 h.
<emphasis id="B955A839FF972E77FCA2FB766B6CFCC3" bold="true" box="[828,841,785,793]" italics="true" pageId="2" pageNumber="97">
<superScript id="7C54D963FF972E77FCA2FB766B6CFCC3" attach="right" box="[828,841,785,793]" fontSize="4" pageId="2" pageNumber="97">⁄⁄</superScript>
</emphasis>
<emphasis id="B955A839FF972E77FCD4FB746B71FCF8" box="[842,852,787,802]" italics="true" pageId="2" pageNumber="97">p</emphasis>
&lt;0.01 (
<emphasis id="B955A839FF972E77FC0AFB746BBBFCF8" box="[916,926,787,802]" italics="true" pageId="2" pageNumber="97">n</emphasis>
= 3).
</paragraph>
</caption>
<paragraph id="8B9E742BFF972E77FE39FD9F6AA9F9D4" blockId="2.[423,652,1528,1550]" box="[423,652,1528,1550]" pageId="2" pageNumber="97">
<superScript id="7C54D963FF972E77FE39FD9F698AF9DF" attach="right" box="[423,431,1528,1541]" fontSize="5" pageId="2" pageNumber="97">1</superScript>
H-
<superScript id="7C54D963FF972E77FE5AFD9F69F6F9DF" attach="left" box="[452,467,1528,1541]" fontSize="5" pageId="2" pageNumber="97">13</superScript>
<collectionCode id="ED30ECEEFF972E77FE46FD9A69C3F9D4" box="[472,486,1533,1550]" country="Denmark" name="University of Copenhagen" pageId="2" pageNumber="97" type="Herbarium">C</collectionCode>
HMBC correlations
</paragraph>
<caption id="DF5E24A3FF972E77FFEFFE4A6B6FF9B3" ID-DOI="http://doi.org/10.5281/zenodo.10490608" ID-Zenodo-Dep="10490608" httpUri="https://zenodo.org/record/10490608/files/figure.png" pageId="2" pageNumber="97" startId="2.[113,139,1581,1595]" targetBox="[348,1265,861,1501]" targetPageId="2" targetType="figure">
<paragraph id="8B9E742BFF972E77FFEFFE4A6B6FF9B3" blockId="2.[113,1500,1580,1641]" pageId="2" pageNumber="97">
<emphasis id="B955A839FF972E77FFEFFE4A6887F9E1" bold="true" box="[113,162,1581,1595]" pageId="2" pageNumber="97">Fig. 2.</emphasis>
Structures of the main compounds (
<emphasis id="B955A839FF972E77FE54FE4A69CDF9E1" bold="true" box="[458,488,1581,1595]" pageId="2" pageNumber="97">1–3</emphasis>
) isolated from the leaves of
<taxonomicName id="4C210FA8FF972E77FD54FE4B6B18F9E6" box="[714,829,1580,1596]" class="Pinopsida" family="Cephalotaxaceae" genus="Cephalotaxus" kingdom="Plantae" order="Pinales" pageId="2" pageNumber="97" phylum="Tracheophyta" rank="species" species="harringtonia">
<emphasis id="B955A839FF972E77FD54FE4B6B18F9E6" box="[714,829,1580,1596]" italics="true" pageId="2" pageNumber="97">C. harringtonia</emphasis>
</taxonomicName>
(
<emphasis id="B955A839FF972E77FCD6FE4A6B71F9E1" bold="true" box="[840,852,1581,1595]" pageId="2" pageNumber="97">A</emphasis>
) biotransformed by the endophytic fungus
<taxonomicName id="4C210FA8FF972E77FB2EFE4A6D20F9E6" box="[1200,1285,1581,1596]" class="Dothideomycetes" family="Didymosphaeriaceae" genus="Paraconiothyrium" kingdom="Fungi" order="Pleosporales" pageId="2" pageNumber="97" phylum="Ascomycota" rank="species" species="variabile">
<emphasis id="B955A839FF972E77FB2EFE4A6D20F9E6" box="[1200,1285,1581,1596]" italics="true" pageId="2" pageNumber="97">P. variabile</emphasis>
</taxonomicName>
and their fragments at
<emphasis id="B955A839FF972E77FA5EFE4A6DF9F9E6" box="[1472,1500,1581,1596]" italics="true" pageId="2" pageNumber="97">m/z</emphasis>
provided by fragmentation of their respective protonated ion [M+H]
<superScript id="7C54D963FF972E77FD0FFE276ABDF993" attach="left" box="[657,664,1600,1609]" fontSize="4" pageId="2" pageNumber="97">+</superScript>
at
<emphasis id="B955A839FF972E77FD2EFE246AE9F988" box="[688,716,1603,1618]" italics="true" pageId="2" pageNumber="97">m/z</emphasis>
579.14, 621.18, 651.19 from CID experiments. Key NMR correlations (HMBC) are also represented for localization of the acetate group of compound (
<emphasis id="B955A839FF972E77FD8CFE3C6A39F9B3" bold="true" box="[530,540,1627,1641]" pageId="2" pageNumber="97">2</emphasis>
) and are identical for compound (
<emphasis id="B955A839FF972E77FCAAFE3C6B1BF9B3" bold="true" box="[820,830,1627,1641]" pageId="2" pageNumber="97">3</emphasis>
).
</paragraph>
</caption>
<paragraph id="8B9E742BFF972E77FFECFED4685BF939" blockId="2.[113,783,1715,1763]" pageId="2" pageNumber="97">
<emphasis id="B955A839FF972E77FFECFED46B2AF91D" box="[114,783,1715,1735]" italics="true" pageId="2" pageNumber="97">
2.3. Preparative scale biotransformation of the isolated compounds
<emphasis id="B955A839FF972E77FD6BFED46B2AF91D" bold="true" box="[757,783,1715,1735]" italics="true" pageId="2" pageNumber="97">1–</emphasis>
</emphasis>
<subSection id="E2AE6DC0FF972E77FFEFFEA86BAFF8C1" pageId="2" pageNumber="97" type="multiple">
<emphasis id="B955A839FF972E77FFEFFEA8685BF939" bold="true" box="[113,126,1743,1763]" italics="true" pageId="2" pageNumber="97">3</emphasis>
<paragraph id="8B9E742BFF972E77FF0FFF6F6BAFF8C1" blockId="2.[113,783,1800,2015]" lastBlockId="2.[831,1501,1716,1819]" pageId="2" pageNumber="97">
Preparative scale biotransformations of the pure glycosylated flavonoids
<emphasis id="B955A839FF972E77FF7FFF436922F8ED" bold="true" box="[225,263,1828,1847]" pageId="2" pageNumber="97">1–3</emphasis>
obtained from the crude extract of
<taxonomicName id="4C210FA8FF972E77FDE3FF446B2AF8ED" box="[637,783,1827,1847]" class="Pinopsida" family="Cephalotaxaceae" genus="Cephalotaxus" kingdom="Plantae" order="Pinales" pageId="2" pageNumber="97" phylum="Tracheophyta" rank="species" species="harringtonia">
<emphasis id="B955A839FF972E77FDE3FF446B2AF8ED" box="[637,783,1827,1847]" italics="true" pageId="2" pageNumber="97">C. harringtonia</emphasis>
</taxonomicName>
were performed. Compounds
<emphasis id="B955A839FF972E77FE3BFF2769EEF889" bold="true" box="[421,459,1856,1875]" pageId="2" pageNumber="97">1–3</emphasis>
were added to
<taxonomicName id="4C210FA8FF972E77FDEFFF586AFAF889" box="[625,735,1855,1875]" class="Dothideomycetes" family="Didymosphaeriaceae" genus="Paraconiothyrium" kingdom="Fungi" order="Pleosporales" pageId="2" pageNumber="97" phylum="Ascomycota" rank="species" species="variabile">
<emphasis id="B955A839FF972E77FDEFFF586AFAF889" box="[625,735,1855,1875]" italics="true" pageId="2" pageNumber="97">P. variabile</emphasis>
</taxonomicName>
in a phosphate buffer. In all cases, large decreases of the substrate concentrations were observed by
<collectionCode id="ED30ECEEFF972E77FE3BFF1F69FFF851" box="[421,474,1912,1931]" pageId="2" pageNumber="97">HPLC</collectionCode>
monitoring after 24 h of inoculation. The resulting products were isolated by chromatography before NMR and
<collectionCode id="ED30ECEEFF972E77FE82FFD76919F819" box="[284,316,1968,1987]" country="Italy" lsid="urn:lsid:biocol.org:col:14396" name="Herbarium Messanaensis, Università di Messina" pageId="2" pageNumber="97" type="Herbarium">MS</collectionCode>
analyses. From these, we saw deglycosylation reactions in all cases, leading to two different aglycones. Indeed, from flavonoids
<emphasis id="B955A839FF972E77FC72FED36BDCF91D" bold="true" box="[1004,1017,1716,1735]" pageId="2" pageNumber="97">1</emphasis>
and
<emphasis id="B955A839FF972E77FBAAFED36C64F91D" bold="true" box="[1076,1089,1716,1735]" pageId="2" pageNumber="97">2</emphasis>
, apigenin (
<emphasis id="B955A839FF972E77FB25FED36CEDF91D" bold="true" box="[1211,1224,1716,1735]" pageId="2" pageNumber="97">4</emphasis>
) was characterized while, from flavonoid
<emphasis id="B955A839FF972E77FC77FEB76BD3F939" bold="true" box="[1001,1014,1744,1763]" pageId="2" pageNumber="97">3</emphasis>
, the small amount of product (
<emphasis id="B955A839FF972E77FAC3FEB66D4FF93E" bold="true" box="[1373,1386,1745,1764]" pageId="2" pageNumber="97">5</emphasis>
) could be assigned as chrysoeriol by comparison with a commercial standard (
<figureCitation id="131A68AEFF972E77FCD9FF6F6B5BF8C1" box="[839,894,1800,1819]" captionStart="Fig" captionStartId="3.[87,113,1976,1990]" captionTargetBox="[337,1218,1182,1945]" captionTargetId="graphics-862@3.[368,759,1720,1923]" captionTargetPageId="3" captionText="Fig. 3. Biotransformations by P. variabile of the glycosylated flavonoids (1–3) and isolated from the crude extract of C. harringtonia and structures of the obtained aglycone moieties (4,5) evidenced by metabolomics." figureDoi="http://doi.org/10.5281/zenodo.10490610" httpUri="https://zenodo.org/record/10490610/files/figure.png" pageId="2" pageNumber="97">Fig. 3</figureCitation>
).
</paragraph>
</subSection>
</paragraph>
<paragraph id="8B9E742BFF972E77FCA1FF586D88F889" blockId="2.[831,1453,1855,1875]" box="[831,1453,1855,1875]" pageId="2" pageNumber="97">
<heading id="D0D6C347FF972E77FCA1FF586D88F889" box="[831,1453,1855,1875]" fontSize="36" level="2" pageId="2" pageNumber="97" reason="3">
<emphasis id="B955A839FF972E77FCA1FF586D88F889" box="[831,1453,1855,1875]" italics="true" pageId="2" pageNumber="97">
2.4. Flavones
<emphasis id="B955A839FF972E77FC5CFF586BEAF889" bold="true" box="[962,975,1855,1875]" italics="true" pageId="2" pageNumber="97">4</emphasis>
and
<emphasis id="B955A839FF972E77FB9FFF586C2BF889" bold="true" box="[1025,1038,1855,1875]" italics="true" pageId="2" pageNumber="97">5</emphasis>
increase the growth of
<taxonomicName id="4C210FA8FF972E77FB6BFF586D88F889" box="[1269,1453,1855,1875]" class="Dothideomycetes" family="Didymosphaeriaceae" genus="Paraconiothyrium" kingdom="Fungi" order="Pleosporales" pageId="2" pageNumber="97" phylum="Ascomycota" rank="subSpecies" species="variabile" subSpecies="hyphae">P. variabile hyphae</taxonomicName>
</emphasis>
</heading>
</paragraph>
<paragraph id="8B9E742BFF972E76FCC0FF1F6A5FFE8F" blockId="2.[831,1501,1912,2015]" lastBlockId="3.[87,757,182,341]" lastPageId="3" lastPageNumber="98" pageId="2" pageNumber="97">
We investigated the respective impact of glycosylated flavonoids (
<emphasis id="B955A839FF972E77FC19FFF36B8BF87D" bold="true" box="[903,942,1940,1959]" pageId="2" pageNumber="97">1–3</emphasis>
) and aglycones (
<emphasis id="B955A839FF972E77FBFAFFF36C54F87D" bold="true" box="[1124,1137,1940,1959]" pageId="2" pageNumber="97">4</emphasis>
,
<emphasis id="B955A839FF972E77FBE8FFF36CA6F87D" bold="true" box="[1142,1155,1940,1959]" pageId="2" pageNumber="97">5</emphasis>
) on the germination and hyphal growth of the spores of
<taxonomicName id="4C210FA8FF972E77FBABFFC86C84F819" box="[1077,1185,1967,1987]" class="Dothideomycetes" family="Didymosphaeriaceae" genus="Paraconiothyrium" kingdom="Fungi" order="Pleosporales" pageId="2" pageNumber="97" phylum="Ascomycota" rank="species" species="variabile">
<emphasis id="B955A839FF972E77FBABFFC86C84F819" box="[1077,1185,1967,1987]" italics="true" pageId="2" pageNumber="97">P. variabile</emphasis>
</taxonomicName>
. While no effect was observed on spore germination in any case (data not shown), we clearly observed (
<figureCitation id="131A68AEFF962E76FF5AF8D168D9FF13" box="[196,252,182,201]" captionStart="Fig" captionStartId="4.[113,139,770,784]" captionTargetBox="[349,1256,182,739]" captionTargetId="figure-696@4.[348,1257,181,740]" captionTargetPageId="4" captionText="Fig. 4. Hyphal length (mm) from the spores of P. variabile in presence of glycosylated flavonoids (1–3) and aglycones (4–5) after 24 h and compared to control DMSO. ⁄⁄⁄p &lt;0.001 (n = 3)." figureDoi="http://doi.org/10.5281/zenodo.10490612" httpUri="https://zenodo.org/record/10490612/files/figure.png" pageId="3" pageNumber="98">Fig. 4</figureCitation>
) that after 24 h, the two aglycones apigenin (
<emphasis id="B955A839FF962E76FD7EF8D16AC8FF13" bold="true" box="[736,749,182,201]" pageId="3" pageNumber="98">4</emphasis>
) and, mostly, chrysoeriol (
<emphasis id="B955A839FF962E76FEC0F8B5694EFF3F" bold="true" box="[350,363,210,229]" pageId="3" pageNumber="98">5</emphasis>
) caused a significant improvement of the hyphal growth compared to the corresponding glycosylated compounds (
<emphasis id="B955A839FF962E76FF42F96D6927FEC7" bold="true" box="[220,258,266,285]" pageId="3" pageNumber="98">1–3</emphasis>
). No inhibition of spore germination and hyphal growth of the spore of
<taxonomicName id="4C210FA8FF962E76FEA6F9426987FEE3" box="[312,418,293,313]" class="Dothideomycetes" family="Didymosphaeriaceae" genus="Paraconiothyrium" kingdom="Fungi" order="Pleosporales" pageId="3" pageNumber="98" phylum="Ascomycota" rank="species" species="variabile">
<emphasis id="B955A839FF962E76FEA6F9426987FEE3" box="[312,418,293,313]" italics="true" pageId="3" pageNumber="98">P. variabile</emphasis>
</taxonomicName>
was observed with either the glycosylated flavonoids, or the corresponding flavonoids.
</paragraph>
</subSubSection>
</treatment>
</document>
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