243 lines
32 KiB
XML
243 lines
32 KiB
XML
<document id="8DE3B0599AF3F2CA2BB6332E0CE7F43C" ID-DOI="10.1016/j.phytochem.2019.04.013" ID-ISSN="1873-3700" ID-Zenodo-Dep="10483032" IM.bibliography_approvedBy="felipe" IM.illustrations_approvedBy="valdenar" IM.materialsCitations_approvedBy="felipe" IM.metadata_approvedBy="felipe" IM.tables_approvedBy="valdenar" IM.taxonomicNames_approvedBy="valdenar" IM.treatments_approvedBy="valdenar" checkinTime="1704933699037" checkinUser="felipe" docAuthor="Zhang, Chi, Chen, Xinlu, Crandall-Stotler, Barbara, Qian, Ping, Köllner, Tobias G., Guo, Hong & Chen, Feng" docDate="2019" docId="03AB0102996DE01BAB4B08D9FCF93A23" docLanguage="en" docName="Phytochemistry.164.50-59.pdf" docOrigin="Phytochemistry 164" docSource="http://dx.doi.org/10.1016/j.phytochem.2019.04.013" docStyle="DocumentStyle:9E596C34F4E94307D29315B03ACE1007.6:Phytochemistry.2014-2019.journal_article" docStyleId="9E596C34F4E94307D29315B03ACE1007" docStyleName="Phytochemistry.2014-2019.journal_article" docStyleVersion="6" docTitle="Conocephalum salebrosum" docType="treatment" docVersion="1" lastPageNumber="53" masterDocId="FF92797A996CE018AB2F0D2FFFF8385C" masterDocTitle="Biosynthesis of methyl (E) - cinnamate in the liverwort Conocephalum salebrosum and evolution of cinnamic acid methyltransferase" masterLastPageNumber="59" masterPageNumber="50" pageNumber="51" updateTime="1706537424403" updateUser="valdenar">
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<mods:title id="20D3E7483DFFD625EB987FDB1522E159">Biosynthesis of methyl (E) - cinnamate in the liverwort Conocephalum salebrosum and evolution of cinnamic acid methyltransferase</mods:title>
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<mods:namePart id="5B22DD3E1EE433D242F4CC6B69CD69B0">Zhang, Chi</mods:namePart>
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<mods:affiliation id="9D7615AC974304BD500EE4C5D43E7EE9">∗ & Department of Plant Sciences, University of Tennessee, Knoxville, TN 37996, USA</mods:affiliation>
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<mods:namePart id="0FF3AC074DFFB88E4A71651E4205C965">Chen, Xinlu</mods:namePart>
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<mods:namePart id="B5B3C102FBCBB1376C1481ABCA3CDDD2">Crandall-Stotler, Barbara</mods:namePart>
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<mods:namePart id="6E15A679E16EDD150E298C8BC35E7671">Qian, Ping</mods:namePart>
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<mods:namePart id="E6AEEAEE86C4080E06E6B63D967BF099">Köllner, Tobias G.</mods:namePart>
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<mods:namePart id="3F79FB6BB3F8154C2656ABA8D2162F87">Guo, Hong</mods:namePart>
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<mods:namePart id="0BFB5E658FBFF9E4E01D5D1BB1FB5031">Chen, Feng</mods:namePart>
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<mods:title id="316F079C6290F892A2456E509C1BAE17">Phytochemistry</mods:title>
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<mods:date id="2555A6F7349193B1CDCBADCDD3F1B6F9">2019</mods:date>
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<mods:number id="21F145007EE09E665908CF6E1266B919">2019-08-31</mods:number>
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<mods:number id="DFA0D025BFA700AF215C559DD91720A0">164</mods:number>
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<mods:url id="5EFACDDA0B02C1CA22D3AF2C6566A375">http://dx.doi.org/10.1016/j.phytochem.2019.04.013</mods:url>
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<mods:classification id="DAEC33398422BA8E1DEC7525154EB5A5">journal article</mods:classification>
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<mods:identifier id="2A5D3A537FF136BA4D53A7701EDDB14A" type="DOI">10.1016/j.phytochem.2019.04.013</mods:identifier>
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<mods:identifier id="1E1B616751743BAD1E45BE80755813B4" type="ISSN">1873-3700</mods:identifier>
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<treatment id="03AB0102996DE01BAB4B08D9FCF93A23" LSID="urn:lsid:plazi:treatment:03AB0102996DE01BAB4B08D9FCF93A23" httpUri="http://treatment.plazi.org/id/03AB0102996DE01BAB4B08D9FCF93A23" lastPageId="3" lastPageNumber="53" pageId="1" pageNumber="51">
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<subSubSection id="C318E39F996DE019AB4B08D9FD033E55" box="[100,763,1526,1545]" pageId="1" pageNumber="51" type="nomenclature">
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<paragraph id="8BBDB014996DE019AB4B08D9FD033E55" blockId="1.[100,763,1526,1545]" box="[100,763,1526,1545]" pageId="1" pageNumber="51">
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<heading id="D0F50778996DE019AB4B08D9FD033E55" bold="true" box="[100,763,1526,1545]" centered="true" fontSize="36" level="1" pageId="1" pageNumber="51" reason="1">
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<emphasis id="B9766C06996DE019AB4B08D9FD033E55" bold="true" box="[100,763,1526,1545]" italics="true" pageId="1" pageNumber="51">
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2.1. Production of axenic culture of
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<taxonomicName id="4C02CB97996DE019AA9F08D9FDD53E55" ID-CoL="5ZSN9" authority="Szweyk., Buczk. & Odrzyk" box="[432,557,1526,1545]" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="salebrosum">C. salebrosum</taxonomicName>
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and chemical profiling
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</emphasis>
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</heading>
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</paragraph>
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</subSubSection>
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<subSubSection id="C318E39F996DE01BABAA0B01FCF93A23" lastPageId="3" lastPageNumber="53" pageId="1" pageNumber="51" type="description">
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<paragraph id="8BBDB014996DE019ABAA0B01FC733C95" blockId="1.[100,770,1582,1992]" lastBlockId="1.[818,1488,1038,1225]" pageId="1" pageNumber="51">
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The sample collected from
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<collectingRegion id="49C67EF6996DE019AAA90B01FE3F3E1D" box="[390,455,1582,1601]" country="United States of America" name="Illinois" pageId="1" pageNumber="51">Illinois</collectingRegion>
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(IL) morphologically matched
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<taxonomicName id="4C02CB97996DE019A9C10B01FF3F3E00" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="salebrosum">
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<emphasis id="B9766C06996DE019A9C10B01FF3F3E00" bold="true" italics="true" pageId="1" pageNumber="51">C. salebrosum</emphasis>
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</taxonomicName>
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, a segregate species of
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<taxonomicName id="4C02CB97996DE019AAEE0B65FF663E25" authority="(Szweykowski et al., 2005)" baseAuthorityName="Szweykowski" baseAuthorityYear="2005" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="conicum">
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<emphasis id="B9766C06996DE019AAEE0B65FDD23E01" bold="true" box="[449,554,1610,1629]" italics="true" pageId="1" pageNumber="51">C. conicum</emphasis>
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(
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<bibRefCitation id="EF93CDE5996DE019A9110B65FF6C3E24" author="Szweykowski, J. & Buczkowska, K. & Odrzykoski, I." pageId="1" pageNumber="51" pagination="133 - 158" refId="ref11373" refString="Szweykowski, J., Buczkowska, K., Odrzykoski, I., 2005. Conocephalum salebrosum (Marchantiopsida, Conocephalaceae) - a new Holarctic liverwort species. Plant Systemat. Evol. 253, 133 - 158." type="journal article" year="2005">Szweykowski et al., 2005</bibRefCitation>
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)
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</taxonomicName>
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. This population is the chemotype with methyl (
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<emphasis id="B9766C06996DE019A9430B49FD803E25" bold="true" box="[620,632,1638,1657]" italics="true" pageId="1" pageNumber="51">E</emphasis>
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)-cinnamate as the dominant compound described by
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<bibRefCitation id="EF93CDE5996DE019AACC0BAEFD593EC8" author="Wood, W. F. & Lancaster, W. C. & Fisher, C. O. & Stotler, R. E." box="[483,673,1665,1685]" pageId="1" pageNumber="51" pagination="241 - 242" refId="ref11597" refString="Wood, W. F., Lancaster, W. C., Fisher, C. O., Stotler, R. E., 1996. (E) - methyl cinnamate: the major volatile from some populations of the liverwort, Conocephalum conicum. Phytochemistry 42, 241 - 242." type="journal article" year="1996">Wood et al. (1996)</bibRefCitation>
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, and was therefore chosen as the model chemotype for most of the experiments in this study. Live plants were collected in the field and grown in axenic culture. Chemical profiling analysis was performed using gas chromatography–mass spectrometry (GC-MS) to determine the volatile components in the axenically grown gametophytes of this species of the
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<taxonomicName id="4C02CB97996DE019A9C10A22FF563F60" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="conicum">
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<emphasis id="B9766C06996DE019A9C10A22FF563F60" bold="true" italics="true" pageId="1" pageNumber="51">C. conicum</emphasis>
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</taxonomicName>
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complex. From organic extraction, a total of 10 volatiles were detected (
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<figureCitation id="1339AC91996DE019ABEB0A6AFEF63F04" box="[196,270,1861,1880]" captionStart="Fig" captionStartId="1.[818,848,822,839]" captionTargetBox="[823,1481,152,800]" captionTargetPageId="1" captionText="Fig. 1. Chemical analysis of C. salebrosum. (A) GC chromatogram of the organic extraction of C. salebrosum. 1*, sabinene; 2, 1-octanol; 3, 1-octenyl acetate; 4*, methyl (E)-cinnamate; 5, unidentified sesquiterpene hydrocarbon; 6, 10-epi- cubebol; 7, 1-epi-cubenol; 8, 1-octadecyne; 9, (E)-2-tetradecen-1-ol; 10, phytol. (B) GC chromatogram of headspace collection of C. salebrosum. 1*, sabinene; 4*, methyl (E)-cinnamate; 5, unidentified sesquiterpene hydrocarbon. “*” indicates compounds that were verified by authentic standards." figureDoi="http://doi.org/10.5281/zenodo.10483034" httpUri="https://zenodo.org/record/10483034/files/figure.png" pageId="1" pageNumber="51">Fig. 1A</figureCitation>
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), of which methyl (
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<emphasis id="B9766C06996DE019AAFF0A6AFE243F04" bold="true" box="[464,476,1861,1880]" italics="true" pageId="1" pageNumber="51">E</emphasis>
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)-cinnamate was the dominant component, accounting for 50.5% of the total volatiles. The monoterpene sabinene was the second most abundant with a concentration of 20% of that of methyl (
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<emphasis id="B9766C06996DE019AA130AB6FEB03FF0" bold="true" box="[316,328,1945,1964]" italics="true" pageId="1" pageNumber="51">E</emphasis>
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)-cinnamate. The thallus of
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<taxonomicName id="4C02CB97996DE019A9670AB7FD3C3FF7" box="[584,708,1944,1963]" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="salebrosum">
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<emphasis id="B9766C06996DE019A9670AB7FD3C3FF7" bold="true" box="[584,708,1944,1963]" italics="true" pageId="1" pageNumber="51">C. salebrosum</emphasis>
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</taxonomicName>
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, when growing in the wild, possesses a characteristic aromatic odor. To determine whether
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<taxonomicName id="4C02CB97996DE019A8C20921FB923C7D" box="[1005,1130,1038,1057]" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="salebrosum">
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<emphasis id="B9766C06996DE019A8C20921FB923C7D" bold="true" box="[1005,1130,1038,1057]" italics="true" pageId="1" pageNumber="51">C. salebrosum</emphasis>
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</taxonomicName>
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grown axenically on culture medium emits volatiles, we performed headspace collection of explants of
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<taxonomicName id="4C02CB97996DE019AE930905FC6D3C05" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="salebrosum">
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<emphasis id="B9766C06996DE019AE930905FC6D3C05" bold="true" italics="true" pageId="1" pageNumber="51">C. salebrosum</emphasis>
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</taxonomicName>
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, which was analyzed by GC-MS. Three dominant compounds were detected (
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<figureCitation id="1339AC91996DE019A8EA094DFBF33C29" box="[965,1035,1122,1141]" captionStart="Fig" captionStartId="1.[818,848,822,839]" captionTargetBox="[823,1481,152,800]" captionTargetPageId="1" captionText="Fig. 1. Chemical analysis of C. salebrosum. (A) GC chromatogram of the organic extraction of C. salebrosum. 1*, sabinene; 2, 1-octanol; 3, 1-octenyl acetate; 4*, methyl (E)-cinnamate; 5, unidentified sesquiterpene hydrocarbon; 6, 10-epi- cubebol; 7, 1-epi-cubenol; 8, 1-octadecyne; 9, (E)-2-tetradecen-1-ol; 10, phytol. (B) GC chromatogram of headspace collection of C. salebrosum. 1*, sabinene; 4*, methyl (E)-cinnamate; 5, unidentified sesquiterpene hydrocarbon. “*” indicates compounds that were verified by authentic standards." figureDoi="http://doi.org/10.5281/zenodo.10483034" httpUri="https://zenodo.org/record/10483034/files/figure.png" pageId="1" pageNumber="51">Fig. 1B</figureCitation>
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). All of them were also detected by organic extraction. Similar to organic extraction, methyl (
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<emphasis id="B9766C06996DE019AE2F0951FAF43CCD" bold="true" box="[1280,1292,1150,1169]" italics="true" pageId="1" pageNumber="51">E</emphasis>
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)-cinnamate was the most abundant compound in the volatile bouquet of
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<taxonomicName id="4C02CB97996DE019AE6109B5FA283CF0" box="[1358,1488,1177,1197]" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="salebrosum">
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<emphasis id="B9766C06996DE019AE6109B5FA283CF0" bold="true" box="[1358,1488,1177,1197]" italics="true" pageId="1" pageNumber="51">C. salebrosum</emphasis>
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</taxonomicName>
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(
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<figureCitation id="1339AC91996DE019A815099AFC783C95" box="[826,896,1205,1225]" captionStart="Fig" captionStartId="1.[818,848,822,839]" captionTargetBox="[823,1481,152,800]" captionTargetPageId="1" captionText="Fig. 1. Chemical analysis of C. salebrosum. (A) GC chromatogram of the organic extraction of C. salebrosum. 1*, sabinene; 2, 1-octanol; 3, 1-octenyl acetate; 4*, methyl (E)-cinnamate; 5, unidentified sesquiterpene hydrocarbon; 6, 10-epi- cubebol; 7, 1-epi-cubenol; 8, 1-octadecyne; 9, (E)-2-tetradecen-1-ol; 10, phytol. (B) GC chromatogram of headspace collection of C. salebrosum. 1*, sabinene; 4*, methyl (E)-cinnamate; 5, unidentified sesquiterpene hydrocarbon. “*” indicates compounds that were verified by authentic standards." figureDoi="http://doi.org/10.5281/zenodo.10483034" httpUri="https://zenodo.org/record/10483034/files/figure.png" pageId="1" pageNumber="51">Fig. 1B</figureCitation>
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).
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</paragraph>
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<paragraph id="8BBDB014996DE019A81D09D4FC813D75" blockId="1.[818,1473,1274,1294]" lastBlockId="1.[818,889,1302,1321]" pageId="1" pageNumber="51">
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<emphasis id="B9766C06996DE019A81D09D4FC813D75" bold="true" italics="true" pageId="1" pageNumber="51">
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2.2. Isolation of SABATH genes from
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<taxonomicName id="4C02CB97996DE019AFA509D4FAF03D51" ID-CoL="5ZSN9" box="[1162,1288,1274,1294]" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="salebrosum">C. salebrosum</taxonomicName>
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and gene expression
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<heading id="D0F50778996DE019A81D0839FC813D75" box="[818,889,1302,1321]" fontSize="8" level="3" pageId="1" pageNumber="51" reason="8">analysis</heading>
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</emphasis>
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</paragraph>
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<paragraph id="8BBDB014996DE019A87C0860FC2D3F28" blockId="1.[818,1488,1358,1991]" pageId="1" pageNumber="51">
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We hypothesized that in the
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<taxonomicName id="4C02CB97996DE019AF480860FB1D3D3D" box="[1127,1253,1358,1378]" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="salebrosum">
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<emphasis id="B9766C06996DE019AF480860FB1D3D3D" bold="true" box="[1127,1253,1358,1378]" italics="true" pageId="1" pageNumber="51">C. salebrosum</emphasis>
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</taxonomicName>
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methyl (
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<emphasis id="B9766C06996DE019AE130860FAB03D3E" bold="true" box="[1340,1352,1359,1378]" italics="true" pageId="1" pageNumber="51">E</emphasis>
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)-cinnamate is synthesized by the action of SABATH methyltransferase. To identify
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<emphasis id="B9766C06996DE019A81D08A9FC7A3DC5" bold="true" box="[818,898,1414,1433]" italics="true" pageId="1" pageNumber="51">SABATH</emphasis>
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genes, we analyzed the transcriptome of
|
||
<taxonomicName id="4C02CB97996DE019AE3F08A9FA8C3DC5" box="[1296,1396,1414,1433]" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="conicum">
|
||
<emphasis id="B9766C06996DE019AE3F08A9FA8C3DC5" bold="true" box="[1296,1396,1414,1433]" italics="true" pageId="1" pageNumber="51">C. conicum</emphasis>
|
||
</taxonomicName>
|
||
prepared from vegetative thalli (https://sites.google.com/a/ualberta.ca/onekp), which was produced by the 1 KP consortium (
|
||
<bibRefCitation id="EF93CDE5996DE019AFCC0891FA643D8D" author="Matasci, N. & Hung, L. & Yan, Z. & Carpenter, E. J. & Wickett, N. J. & Mirarab, S. & Nguyen, N. & Warnow, T. & Ayyampalayam, S. & Barker, M. & Burleigh, J. G. & Gitzendanner, M. A. & Wafula, E. & Der, J. P. & dePamphilis, C. W. & Roure, B. & Philippe, H. & Ruhfel, B. R. & Miles, N. W. & Graham, S. W. & Mathews, S. & Surek, B. & Melkonian, M. & Soltis, D. E. & Soltis, P. S. & Rothfels, C. & Pokorny, L. & Shaw, J. A. & DeGironimo, L. & Stevenson, D. W. & Villarreal, J. C. & Chen, T. & Kutchan, T. M. & Rolf, M. & Baucom, R. S. & Deyholos, M. K. & Samudrala, R. & Tian, Z. & Wu, X. & Sun, X. & Zhang, Y. & Wang, J. & Leebens-Mack, J. & Wong, G. K." box="[1251,1436,1470,1490]" pageId="1" pageNumber="51" pagination="17" refId="ref10566" refString="Matasci, N., Hung, L., Yan, Z., Carpenter, E. J., Wickett, N. J., Mirarab, S., Nguyen, N., Warnow, T., Ayyampalayam, S., Barker, M., Burleigh, J. G., Gitzendanner, M. A., Wafula, E., Der, J. P., dePamphilis, C. W., Roure, B., Philippe, H., Ruhfel, B. R., Miles, N. W., Graham, S. W., Mathews, S., Surek, B., Melkonian, M., Soltis, D. E., Soltis, P. S., Rothfels, C., Pokorny, L., Shaw, J. A., DeGironimo, L., Stevenson, D. W., Villarreal, J. C., Chen, T., Kutchan, T. M., Rolf, M., Baucom, R. S., Deyholos, M. K., Samudrala, R., Tian, Z., Wu, X., Sun, X., Zhang, Y., Wang, J., Leebens-Mack, J., Wong, G. K., 2014. Data access for the 1,000 Plants (1 KP) project. GigaScience 3, 17." type="journal article" year="2014">Matasci et al., 2014</bibRefCitation>
|
||
). Via a HMMR search (
|
||
<bibRefCitation id="EF93CDE5996DE019A8F408F5FB843DB1" author="Finn, R. D. & Clements, J. & Eddy, S. R." box="[987,1148,1498,1517]" pageId="1" pageNumber="51" pagination="37" refId="ref9119" refString="Finn, R. D., Clements, J., Eddy, S. R., 2011. HMMER web server: interactive sequence similarity searching. Nucleic Acids Res. 39, W 29 - W 37." type="journal article" year="2011">Finn et al., 2011</bibRefCitation>
|
||
) using Methyltransf_7 Pfam profile (
|
||
<bibRefCitation id="EF93CDE5996DE019A81508D9FC2D3E55" author="Finn, R. D. & Coggill, P. & Eberhardt, R. Y. & Eddy, S. R. & Mistry, J. & Mitchell, A. L. & Potter, S. C. & Punta, M. & Qureshi, M. & Sangrador-Vegas, A. & Salazar, G. A. & Tate, J. & Bateman, A." box="[826,981,1526,1545]" pageId="1" pageNumber="51" pagination="285" refId="ref9161" refString="Finn, R. D., Coggill, P., Eberhardt, R. Y., Eddy, S. R., Mistry, J., Mitchell, A. L., Potter, S. C., Punta, M., Qureshi, M., Sangrador-Vegas, A., Salazar, G. A., Tate, J., Bateman, A., 2015. The Pfam protein families database: towards a more sustainable future. Nucleic Acids Res. 44, D 279 - D 285." type="journal article" year="2015">Finn et al., 2015</bibRefCitation>
|
||
) as query, a total of nine unigenes encoding SABATH proteins were identified. Six of them appear to be in full-length when compared to other SABATH proteins and their corresponding orthologs in
|
||
<taxonomicName id="4C02CB97996DE019A8630B65FC323E00" box="[844,970,1609,1629]" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="salebrosum">
|
||
<emphasis id="B9766C06996DE019A8630B65FC323E00" bold="true" box="[844,970,1609,1629]" italics="true" pageId="1" pageNumber="51">C. salebrosum</emphasis>
|
||
</taxonomicName>
|
||
were designated CsSABATH1-6 (
|
||
<tableCitation id="C68085AF996DE019AE270B65FAA33E01" box="[1288,1371,1610,1629]" captionStart="Table 1" captionStartId="3.[100,150,1537,1553]" captionTargetPageId="3" captionText="Table 1 The relative activities of CsSABATH4 and CsSABATH6 from C. salebrosum with ten carboxyl acids." pageId="1" pageNumber="51">Table S1</tableCitation>
|
||
). Using RT-PCR, full-length cDNAs for all six CsSABATH genes were cloned from the thallus of the axenically grown
|
||
<taxonomicName id="4C02CB97996DE019AFBD0BAEFAE93EC8" box="[1170,1297,1665,1684]" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="salebrosum">
|
||
<emphasis id="B9766C06996DE019AFBD0BAEFAE93EC8" bold="true" box="[1170,1297,1665,1684]" italics="true" pageId="1" pageNumber="51">C. salebrosum</emphasis>
|
||
</taxonomicName>
|
||
. The lengths of six proteins range from 383 to 443 amino acids (
|
||
<figureCitation id="1339AC91996DE019AFCA0BB2FAE43EEC" box="[1253,1308,1693,1712]" captionStart="Fig" captionStartId="2.[100,130,1910,1927]" captionTargetBox="[113,1475,153,1886]" captionTargetId="figure-1@2.[112,1476,152,1887]" captionTargetPageId="2" captionText="Fig. 2. Sequence alignment of CcSABATHs with ObCCMT1 and CbSAMT. Conserved residues are in shade with the more conserved the darker. Residues indicated with “&” are SAM/SAH-binding residues. Residues indicated with “*” are residues that interact with the carboxyl moiety of substrate. Residues indicated with “#” interact with the aromatic moiety of substrate and are important for substrate selectivity. CcSABATHs, C. conicum sensu latu SABATHs; ObCCMT1, Ocimum basilicum cinnamate/p-coumarate carboxyl methyltransferase 1; CbSAMT, Clarkia breweri salicylic acid methyltransferase." figureDoi="http://doi.org/10.5281/zenodo.10483036" httpUri="https://zenodo.org/record/10483036/files/figure.png" pageId="1" pageNumber="51">Fig. 2</figureCitation>
|
||
). Sequence similarities among CsSABATHs range from 37% to 52%. Their similarities to ObCCMT1, one known CAMT from basil, are between 36% and 48%. The amino acid residues for binding the methyl donor SAM among CsSABATHs and ObCCMT1 are identical. The residues that interact with the carboxyl moiety of the substrate are also conserved. However, the residues that interact with the aromatic moiety are significantly different (
|
||
<figureCitation id="1339AC91996DE019A8BE0A4EFC3F3F28" box="[913,967,1889,1908]" captionStart="Fig" captionStartId="2.[100,130,1910,1927]" captionTargetBox="[113,1475,153,1886]" captionTargetId="figure-1@2.[112,1476,152,1887]" captionTargetPageId="2" captionText="Fig. 2. Sequence alignment of CcSABATHs with ObCCMT1 and CbSAMT. Conserved residues are in shade with the more conserved the darker. Residues indicated with “&” are SAM/SAH-binding residues. Residues indicated with “*” are residues that interact with the carboxyl moiety of substrate. Residues indicated with “#” interact with the aromatic moiety of substrate and are important for substrate selectivity. CcSABATHs, C. conicum sensu latu SABATHs; ObCCMT1, Ocimum basilicum cinnamate/p-coumarate carboxyl methyltransferase 1; CbSAMT, Clarkia breweri salicylic acid methyltransferase." figureDoi="http://doi.org/10.5281/zenodo.10483036" httpUri="https://zenodo.org/record/10483036/files/figure.png" pageId="1" pageNumber="51">Fig. 2</figureCitation>
|
||
).
|
||
</paragraph>
|
||
<paragraph id="8BBDB014996DE019A87C0A53FA373F9B" blockId="1.[818,1488,1358,1991]" pageId="1" pageNumber="51">
|
||
With six
|
||
<emphasis id="B9766C06996DE019A8830A53FC043FD3" bold="true" box="[940,1020,1916,1935]" italics="true" pageId="1" pageNumber="51">SABATH</emphasis>
|
||
genes cloned from
|
||
<taxonomicName id="4C02CB97996DE019AF900A53FAC63FD3" box="[1215,1342,1916,1935]" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="1" pageNumber="51" phylum="Marchantiophyta" rank="species" species="salebrosum">
|
||
<emphasis id="B9766C06996DE019AF900A53FAC63FD3" bold="true" box="[1215,1342,1916,1935]" italics="true" pageId="1" pageNumber="51">C. salebrosum</emphasis>
|
||
</taxonomicName>
|
||
, next we compared their expression levels using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis. At 25 cycles of
|
||
</paragraph>
|
||
<caption id="DF7DE09C996EE01AAB4B0A59FBDA3F8F" ID-DOI="http://doi.org/10.5281/zenodo.10483036" ID-Zenodo-Dep="10483036" httpUri="https://zenodo.org/record/10483036/files/figure.png" pageId="2" pageNumber="52" startId="2.[100,130,1910,1927]" targetBox="[113,1475,153,1886]" targetPageId="2" targetType="figure">
|
||
<paragraph id="8BBDB014996EE01AAB4B0A59FBDA3F8F" blockId="2.[100,1488,1910,2004]" pageId="2" pageNumber="52">
|
||
<emphasis id="B9766C06996EE01AAB4B0A59FF663FDB" bold="true" box="[100,158,1910,1927]" pageId="2" pageNumber="52">Fig. 2.</emphasis>
|
||
Sequence alignment of CcSABATHs with ObCCMT1 and CbSAMT. Conserved residues are in shade with the more conserved the darker. Residues indicated with “&” are SAM/SAH-binding residues. Residues indicated with “*” are residues that interact with the carboxyl moiety of substrate. Residues indicated with “#” interact with the aromatic moiety of substrate and are important for substrate selectivity. CcSABATHs,
|
||
<emphasis id="B9766C06996EE01AA8E30A86FB813FE6" bold="true" box="[972,1145,1961,1978]" italics="true" pageId="2" pageNumber="52">
|
||
<taxonomicName id="4C02CB97996EE01AA8E30A86FBAD3FE5" box="[972,1109,1961,1978]" class="Marchantiopsida" family="Conocephalaceae" genus="Conocephalum" kingdom="Plantae" order="Marchantiales" pageId="2" pageNumber="52" phylum="Marchantiophyta" rank="subSpecies" species="conicum" subSpecies="sensu">C. conicum sensu</taxonomicName>
|
||
latu
|
||
</emphasis>
|
||
SABATHs; ObCCMT1,
|
||
<taxonomicName id="4C02CB97996EE01AAE130A86FA373FE6" box="[1340,1487,1961,1978]" class="Magnoliopsida" family="Lamiaceae" genus="Ocimum" kingdom="Plantae" order="Lamiales" pageId="2" pageNumber="52" phylum="Tracheophyta" rank="species" species="basilicum">
|
||
<emphasis id="B9766C06996EE01AAE130A86FA373FE6" bold="true" box="[1340,1487,1961,1978]" italics="true" pageId="2" pageNumber="52">Ocimum basilicum</emphasis>
|
||
</taxonomicName>
|
||
cinnamate/
|
||
<emphasis id="B9766C06996EE01AABE90AECFF2E3F8F" bold="true" box="[198,214,1987,2003]" italics="true" pageId="2" pageNumber="52">p-</emphasis>
|
||
coumarate carboxyl methyltransferase 1; CbSAMT,
|
||
<taxonomicName id="4C02CB97996EE01AA9A20AEDFCF23F8F" box="[653,778,1986,2003]" class="Magnoliopsida" family="Onagraceae" genus="Clarkia" kingdom="Plantae" order="Myrtales" pageId="2" pageNumber="52" phylum="Tracheophyta" rank="species" species="breweri">
|
||
<emphasis id="B9766C06996EE01AA9A20AEDFCF23F8F" bold="true" box="[653,778,1986,2003]" italics="true" pageId="2" pageNumber="52">Clarkia breweri</emphasis>
|
||
</taxonomicName>
|
||
salicylic acid methyltransferase.
|
||
</paragraph>
|
||
</caption>
|
||
<caption id="DF7DE09C996FE01BAB4B0C5EFF213993" ID-DOI="http://doi.org/10.5281/zenodo.10483038" ID-Zenodo-Dep="10483038" httpUri="https://zenodo.org/record/10483038/files/figure.png" pageId="3" pageNumber="53" startId="3.[100,130,369,386]" targetBox="[106,761,152,337]" targetPageId="3" targetType="figure">
|
||
<paragraph id="8BBDB014996FE01BAB4B0C5EFF213993" blockId="3.[100,770,369,463]" pageId="3" pageNumber="53">
|
||
<emphasis id="B9766C06996FE01BAB4B0C5EFF6639DF" bold="true" box="[100,158,369,387]" pageId="3" pageNumber="53">Fig. 3.</emphasis>
|
||
Expression of six
|
||
<emphasis id="B9766C06996FE01BAA130C5DFE6739DF" bold="true" box="[316,415,370,387]" italics="true" pageId="3" pageNumber="53">CsSABATHs</emphasis>
|
||
genes in sterile thalli measured by semiquantitative RT-PCR. Same amount of cDNA was used in each reaction. This experiment was repeated three times with similar results. “M” indicates molecular marker.
|
||
</paragraph>
|
||
</caption>
|
||
<paragraph id="8BBDB014996FE01BAB4B0CD2FCF93A23" blockId="3.[100,770,509,640]" pageId="3" pageNumber="53">
|
||
PCR reactions, transcripts for five of the six genes could be detected from thallus tissue, the exception being
|
||
<emphasis id="B9766C06996FE01BAAD80F36FD903A70" bold="true" box="[503,616,537,556]" italics="true" pageId="3" pageNumber="53">CsSABATH1</emphasis>
|
||
(
|
||
<figureCitation id="1339AC91996FE01BA9540F36FD4C3A70" box="[635,692,537,556]" captionStart="Fig" captionStartId="3.[100,130,369,386]" captionTargetBox="[106,761,152,337]" captionTargetId="graphics-1083@3.[272,761,152,279]" captionTargetPageId="3" captionText="Fig. 3. Expression of six CsSABATHs genes in sterile thalli measured by semiquantitative RT-PCR. Same amount of cDNA was used in each reaction. This experiment was repeated three times with similar results. “M” indicates molecular marker." figureDoi="http://doi.org/10.5281/zenodo.10483038" httpUri="https://zenodo.org/record/10483038/files/figure.png" pageId="3" pageNumber="53">Fig. 3</figureCitation>
|
||
).
|
||
<emphasis id="B9766C06996FE01BA9E30F36FF503A1B" bold="true" italics="true" pageId="3" pageNumber="53">CsSABATH4</emphasis>
|
||
showed the highest level of expression. While
|
||
<emphasis id="B9766C06996FE01BA9480F1BFD203A1B" bold="true" box="[615,728,564,583]" italics="true" pageId="3" pageNumber="53">CsSABATH2</emphasis>
|
||
and
|
||
<emphasis id="B9766C06996FE01BAB4B0F7FFF2D3A3F" bold="true" box="[100,213,592,611]" italics="true" pageId="3" pageNumber="53">CsSABATH6</emphasis>
|
||
showed also higher expression levels.
|
||
<emphasis id="B9766C06996FE01BA94D0F7FFD2C3A3F" bold="true" box="[610,724,592,611]" italics="true" pageId="3" pageNumber="53">CsSABATH3</emphasis>
|
||
and
|
||
<emphasis id="B9766C06996FE01BAB4B0F43FF2D3A23" bold="true" box="[100,213,620,639]" italics="true" pageId="3" pageNumber="53">CsSABATH5</emphasis>
|
||
were found to be expressed at relatively low levels (
|
||
<figureCitation id="1339AC91996FE01BA9900F43FD0C3A23" box="[703,756,620,639]" captionStart="Fig" captionStartId="3.[100,130,369,386]" captionTargetBox="[106,761,152,337]" captionTargetId="graphics-1083@3.[272,761,152,279]" captionTargetPageId="3" captionText="Fig. 3. Expression of six CsSABATHs genes in sterile thalli measured by semiquantitative RT-PCR. Same amount of cDNA was used in each reaction. This experiment was repeated three times with similar results. “M” indicates molecular marker." figureDoi="http://doi.org/10.5281/zenodo.10483038" httpUri="https://zenodo.org/record/10483038/files/figure.png" pageId="3" pageNumber="53">Fig. 3</figureCitation>
|
||
).
|
||
</paragraph>
|
||
</subSubSection>
|
||
</treatment>
|
||
</document> |