460 lines
48 KiB
XML
460 lines
48 KiB
XML
<document id="2076509DECF1C904B66591F98C9F9326" ID-DOI="10.1016/j.phytochem.2020.112283" ID-ISSN="1873-3700" ID-Zenodo-Dep="8294206" IM.bibliography_approvedBy="diego" IM.illustrations_approvedBy="felipe" IM.materialsCitations_approvedBy="diego" IM.metadata_approvedBy="felipe" IM.taxonomicNames_approvedBy="diego" IM.treatments_approvedBy="diego" checkinTime="1693255514846" checkinUser="felipe" docAuthor="Lu, Xiaoxiao, Xie, Chunqin, Wang, Yunxiao, Liu, Yongnan, Han, Jing, Shi, Liang, Zhu, Jing, Yu, Hanshou, Ren, Ang & Zhao, Mingwen" docDate="2020" docId="03D26736FF98FF8D123ADCC8FB0DFA46" docLanguage="en" docName="Phytochemistry.172.112283.pdf" docOrigin="Phytochemistry (112283) 172" docSource="http://dx.doi.org/10.1016/j.phytochem.2020.112283" docStyle="DocumentStyle:F36D69FC8B198FBE91029DF9C24697D3.5:Phytochemistry.2020-.journal_article" docStyleId="F36D69FC8B198FBE91029DF9C24697D3" docStyleName="Phytochemistry.2020-.journal_article" docStyleVersion="5" docTitle="Ganoderma lucidum subsp. contains P. Karst." docType="treatment" docVersion="1" lastPageNumber="3" masterDocId="FFEB1F4EFF99FF8F1108D833FFF6FF94" masterDocTitle="Function of ceramide synthases on growth, ganoderic acid biosynthesis and sphingolipid homeostasis in Ganoderma lucidum" masterLastPageNumber="9" masterPageNumber="1" pageNumber="2" updateTime="1693401629264" updateUser="diego">
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<mods:title id="1E527C7308931655313479CB18EB3F3A">Function of ceramide synthases on growth, ganoderic acid biosynthesis and sphingolipid homeostasis in Ganoderma lucidum</mods:title>
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<mods:namePart id="9E397CFBA6612659B260BDE171A48DBA">Lu, Xiaoxiao</mods:namePart>
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<mods:affiliation id="761DB530945C87AF66BDCAE6480FFEDE">∗ & Key Laboratory of Microbiological Engineering of Agricultural Environment, Ministry of Agriculture, Microbiology Department, College of Life Sciences, Nanjing</mods:affiliation>
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<mods:namePart id="FDCFBFF04C89370C3A7F59B548685FFE">Xie, Chunqin</mods:namePart>
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<mods:namePart id="84DBBE9D8091B423F81E68CA4F9A223A">Wang, Yunxiao</mods:namePart>
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<mods:namePart id="A0D77E37B4AC926E879D99301C6AB4B4">Liu, Yongnan</mods:namePart>
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<mods:namePart id="51D3413C49C10FE9A85609939FF1D333">Han, Jing</mods:namePart>
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<mods:namePart id="F53835E4F018D6DAEFFBB619A9765A8B">Shi, Liang</mods:namePart>
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<mods:namePart id="51AC915A5B0C6C9E7299CD3BE3369A0C">Zhu, Jing</mods:namePart>
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<mods:namePart id="3BF1CFF7B5BCCD71939879A3A03DA1E6">Yu, Hanshou</mods:namePart>
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<mods:namePart id="54CC9A7B58BDB5387FC1A102FB380E96">Ren, Ang</mods:namePart>
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<mods:namePart id="5FE3D04640CF154DD03B17EDC641D152">Zhao, Mingwen</mods:namePart>
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<mods:title id="B3A03695C68B3744D68FCAB4CBB9756C">Phytochemistry</mods:title>
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<mods:part id="73FB27988AE59DF2D1BCEE1C0D7EC204">
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<mods:date id="0D9CEC13B945303B923647435B074205">2020</mods:date>
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<mods:title id="42124CBA17170CBD11AE5B3EE01362C6">112283</mods:title>
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<mods:number id="F6C8688B9FF424D62E78E879ADD36CA9">2020-04-30</mods:number>
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<mods:number id="BAD3C556EDD92C264BEBB3E444AA9D33">172</mods:number>
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<mods:start id="8A0E10899F2F3E344A601283DCE8AE47">1</mods:start>
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<mods:url id="A357958B8C09F3D398E676B915BA75B2">http://dx.doi.org/10.1016/j.phytochem.2020.112283</mods:url>
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<mods:identifier id="295C6EECD459CE2436D3B988072E0A99" type="DOI">10.1016/j.phytochem.2020.112283</mods:identifier>
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<mods:identifier id="5794036103F88258B321F52FAC6DFBDD" type="ISSN">1873-3700</mods:identifier>
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<mods:identifier id="D358C7A0455B25DE17D3EBFA21786195" type="Zenodo-Dep">8294206</mods:identifier>
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<subSubSection id="C36185ABFF98FF8E123ADCC8FB01FA9A" box="[818,1271,1275,1294]" pageId="1" pageNumber="2" type="nomenclature">
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<paragraph id="8BC4D620FF98FF8E123ADCC8FB01FA9A" blockId="1.[818,1271,1275,1294]" box="[818,1271,1275,1294]" pageId="1" pageNumber="2">
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<heading id="D08C614CFF98FF8E123ADCC8FB01FA9A" bold="true" box="[818,1271,1275,1294]" fontSize="36" level="1" pageId="1" pageNumber="2" reason="1">
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<emphasis id="B90F0A32FF98FF8E123ADCC8FB01FA9A" bold="true" box="[818,1271,1275,1294]" italics="true" pageId="1" pageNumber="2">
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2.1.
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<taxonomicName id="4C7BADA3FF98FF8E1268DCC8FC34FA9A" ID-CoL="6JWYZ" ID-ENA="5315" authority="(Curtis) P. Karst." authorityName="P. Karst." baseAuthorityName="Curtis" box="[864,962,1275,1294]" class="Agaricomycetes" family="Ganodermataceae" genus="Ganoderma" kingdom="Fungi" order="Polyporales" pageId="1" pageNumber="2" phylum="Basidiomycota" rank="species" species="lucidum">G. lucidum</taxonomicName>
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contains three ceramide synthases
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</emphasis>
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</heading>
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</paragraph>
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<paragraph id="8BC4D620FF98FF8E125BDD00FC49F8B4" blockId="1.[818,1487,1331,1992]" pageId="1" pageNumber="2">
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On the basis of evidence of ceramide synthases in other fungi, we performed a bioinformatic search for putative ceramide synthases present in
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<taxonomicName id="4C7BADA3FF98FF8E1290DD58FC0DFAEA" box="[920,1019,1387,1406]" class="Agaricomycetes" family="Ganodermataceae" genus="Ganoderma" kingdom="Fungi" order="Polyporales" pageId="1" pageNumber="2" phylum="Basidiomycota" rank="species" species="lucidum">
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<emphasis id="B90F0A32FF98FF8E1290DD58FC0DFAEA" bold="true" box="[920,1019,1387,1406]" italics="true" pageId="1" pageNumber="2">G. lucidum</emphasis>
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</taxonomicName>
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. Our analysis revealed that there are three putative ceramide synthases in
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<taxonomicName id="4C7BADA3FF98FF8E154DDDB4FB5AFA0E" box="[1093,1196,1415,1434]" class="Agaricomycetes" family="Ganodermataceae" genus="Ganoderma" kingdom="Fungi" order="Polyporales" pageId="1" pageNumber="2" phylum="Basidiomycota" rank="species" species="lucidum">
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<emphasis id="B90F0A32FF98FF8E154DDDB4FB5AFA0E" bold="true" box="[1093,1196,1415,1434]" italics="true" pageId="1" pageNumber="2">G. lucidum</emphasis>
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</taxonomicName>
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by homologue alignment by comparing to the proteins from other fungi. They are referred in this study as LAG1, LAG2 and LAG3, respectively. The LAG1, LAG2 and LAG3 had significant homology to other ceramide synthase from
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<emphasis id="B90F0A32FF98FF8E123ADDC5FC6FF9D5" italics="true" pageId="1" pageNumber="2">
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<emphasis id="B90F0A32FF98FF8E123ADDC5FBB0F9B1" bold="true" italics="true" pageId="1" pageNumber="2">
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Basidiomycetes including
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<taxonomicName id="4C7BADA3FF98FF8E151DDDC5FB5EF99D" box="[1045,1192,1526,1545]" class="Agaricomycetes" family="Polyporaceae" genus="Lentinus" kingdom="Fungi" order="Polyporales" pageId="1" pageNumber="2" phylum="Basidiomycota" rank="species" species="tigrinus">Lentinus tigrinus</taxonomicName>
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,
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<taxonomicName id="4C7BADA3FF98FF8E15BADDC5FAB9F99D" box="[1202,1359,1526,1545]" class="Agaricomycetes" family="Physalacriaceae" genus="Armillaria" kingdom="Fungi" order="Agaricales" pageId="1" pageNumber="2" phylum="Basidiomycota" rank="species" species="gallica">Armillaria gallica</taxonomicName>
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,
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<taxonomicName id="4C7BADA3FF98FF8E1452DDC5FC5FF9B1" class="Tremellomycetes" family="Tremellaceae" genus="Cryptococcus" kingdom="Fungi" order="Tremellales" pageId="1" pageNumber="2" phylum="Basidiomycota" rank="species" species="depauperatus">Cryptococcus depauperatus</taxonomicName>
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,
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<taxonomicName id="4C7BADA3FF98FF8E12BBDE21FBE0F9B1" authorityName="Y.Yamada, H.Kawas., Itoh, I.Banno & Nakase" authorityYear="1988" box="[947,1046,1554,1573]" class="Tremellomycetes" family="Tremellaceae" genus="Tsuchiyaea" kingdom="Fungi" order="Tremellales" pageId="1" pageNumber="2" phylum="Basidiomycota" rank="genus">Tsuchiyaea</taxonomicName>
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wing
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</emphasis>
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fi
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<emphasis id="B90F0A32FF98FF8E155BDE21FC6FF9D5" bold="true" italics="true" pageId="1" pageNumber="2">
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eldii,
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<taxonomicName id="4C7BADA3FF98FF8E158CDE21FADDF9B1" box="[1156,1323,1554,1573]" class="Tremellomycetes" family="Tremellaceae" genus="Cryptococcus" kingdom="Fungi" order="Tremellales" pageId="1" pageNumber="2" phylum="Basidiomycota" rank="species" species="gattii">Cryptococcus gattii</taxonomicName>
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and
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<taxonomicName id="4C7BADA3FF98FF8E1452DE21FC6FF9D5" class="Tremellomycetes" family="Tremellaceae" genus="Cryptococcus" kingdom="Fungi" order="Tremellales" pageId="1" pageNumber="2" phylum="Basidiomycota" rank="species" species="neoformans">Cryptococcus neoformans</taxonomicName>
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</emphasis>
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</emphasis>
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. The genomic DNA sequence of the
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<emphasis id="B90F0A32FF98FF8E15E1DE1DFAF9F9D5" bold="true" box="[1257,1295,1582,1601]" italics="true" pageId="1" pageNumber="2">lag1</emphasis>
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gene is 1385 bp and is interrupted by three introns, whereas the spliced transcript of
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<emphasis id="B90F0A32FF98FF8E149CDE79FA4FF9C9" bold="true" box="[1428,1465,1610,1629]" italics="true" pageId="1" pageNumber="2">lag1</emphasis>
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is 1089 bp and encodes a protein containing 362 amino acids. The genomic DNA sequence of the
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<emphasis id="B90F0A32FF98FF8E1550DEB1FB8BF901" bold="true" box="[1112,1149,1666,1685]" italics="true" pageId="1" pageNumber="2">lag2</emphasis>
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gene is composed of 1844 bp gene and contains nine introns, whereas the 1317 bp spliced transcript of
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<emphasis id="B90F0A32FF98FF8E123ADE8AFCA1F958" bold="true" box="[818,855,1721,1740]" italics="true" pageId="1" pageNumber="2">lag2</emphasis>
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encodes a protein consisting of 438 amino acids. The cDNA of
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<emphasis id="B90F0A32FF98FF8E14A1DE8AFA39F958" bold="true" box="[1449,1487,1721,1740]" italics="true" pageId="1" pageNumber="2">lag3</emphasis>
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contains a spliced transcript of 1431 bp encoding 476 amino acids, whereas the
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<emphasis id="B90F0A32FF98FF8E12BCDEC2FC38F890" bold="true" box="[948,974,1777,1796]" italics="true" pageId="1" pageNumber="2">lag</emphasis>
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3 genomic DNA sequence is 2083 bp and contains eleven introns.
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</paragraph>
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<paragraph id="8BC4D620FF98FF8D125BDF1AFA37FF26" blockId="1.[818,1487,1331,1992]" lastBlockId="2.[818,1488,159,402]" lastPageId="2" lastPageNumber="3" pageId="1" pageNumber="2">
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We investigated the divergence of the three CERSs in
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<taxonomicName id="4C7BADA3FF98FF8E1462DF1AFA39F8A8" box="[1386,1487,1833,1852]" class="Agaricomycetes" family="Ganodermataceae" genus="Ganoderma" kingdom="Fungi" order="Polyporales" pageId="1" pageNumber="2" phylum="Basidiomycota" rank="species" species="lucidum">
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<emphasis id="B90F0A32FF98FF8E1462DF1AFA39F8A8" bold="true" box="[1386,1487,1833,1852]" italics="true" pageId="1" pageNumber="2">G. lucidum</emphasis>
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</taxonomicName>
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from other known CERS proteins from animals, plants and fungi over the course of evolution. The entire phylogenetic tree can be divided into three major groups (
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<figureCitation id="1340CAA5FF98FF8E12F5DF4EFBC3F804" box="[1021,1077,1917,1936]" captionStart="Fig" captionStartId="2.[100,130,1274,1291]" captionTargetBox="[106,765,154,1243]" captionTargetId="figure-844@2.[105,766,152,1251]" captionTargetPageId="2" captionText="Fig. 2. G. lucidum harbours three ceramide synthases. Phylogenetic analysis of ceramide synthases in eukaryotes. The phylogenetic tree was conducted using MEGA 6 and can be roughly divided into three major groups: human, plant and fungi. The evolutionary history was inferred by the neighbour-joining method from 1000 replicates using MEGA 6." figureDoi="http://doi.org/10.5281/zenodo.8294210" httpUri="https://zenodo.org/record/8294210/files/figure.png" pageId="1" pageNumber="2">Fig. 2</figureCitation>
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). Interestingly, the LAG1 protein from
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<taxonomicName id="4C7BADA3FF98FF8E14B2DF4EFC8FF838" class="Agaricomycetes" family="Ganodermataceae" genus="Ganoderma" kingdom="Fungi" order="Polyporales" pageId="1" pageNumber="2" phylum="Basidiomycota" rank="species" species="lucidum">
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<emphasis id="B90F0A32FF98FF8E14B2DF4EFC8FF838" bold="true" italics="true" pageId="1" pageNumber="2">G. lucidum</emphasis>
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</taxonomicName>
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, the Cer1 proteins from
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<emphasis id="B90F0A32FF98FF8E1569DFAAFB11F838" bold="true" box="[1121,1255,1945,1964]" italics="true" pageId="1" pageNumber="2">Basidiomycetes</emphasis>
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and the Lac1 (or Bar1p) proteins from
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<emphasis id="B90F0A32FF98FF8E12B1DF86FBDEF85C" bold="true" box="[953,1064,1973,1992]" italics="true" pageId="1" pageNumber="2">Ascomycetes</emphasis>
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are clustered together. The LAG2 and LAG3 proteins from
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<taxonomicName id="4C7BADA3FF9BFF8D11E5DDACFEA7FA26" box="[237,337,1439,1458]" class="Agaricomycetes" family="Ganodermataceae" genus="Ganoderma" kingdom="Fungi" order="Polyporales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="lucidum">
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<emphasis id="B90F0A32FF9BFF8D11E5DDACFEA7FA26" bold="true" box="[237,337,1439,1458]" italics="true" pageId="2" pageNumber="3">G. lucidum</emphasis>
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</taxonomicName>
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, the Cer2 and Cer3 proteins from
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<emphasis id="B90F0A32FF9BFF8D1397DDACFF78FA5A" bold="true" italics="true" pageId="2" pageNumber="3">Basidiomycetes</emphasis>
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and the Lag1 proteins in
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<emphasis id="B90F0A32FF9BFF8D1094DD88FDFDFA5A" bold="true" box="[412,523,1467,1486]" italics="true" pageId="2" pageNumber="3">Ascomycetes</emphasis>
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grouped into a separate cluster. In addition, LAG1, LAG2 and LAG3 from
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<taxonomicName id="4C7BADA3FF9BFF8D133ADDE4FD62FA7E" box="[562,660,1495,1514]" class="Agaricomycetes" family="Ganodermataceae" genus="Ganoderma" kingdom="Fungi" order="Polyporales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="lucidum">
|
||
<emphasis id="B90F0A32FF9BFF8D133ADDE4FD62FA7E" bold="true" box="[562,660,1495,1514]" italics="true" pageId="2" pageNumber="3">G. lucidum</emphasis>
|
||
</taxonomicName>
|
||
exhibited a high degree of sequence identity with other ceramide synthases from
|
||
<emphasis id="B90F0A32FF9BFF8D116CDE3CFF1CF9B6" bold="true" box="[100,234,1551,1570]" italics="true" pageId="2" pageNumber="3">Basidiomycetes</emphasis>
|
||
including
|
||
<taxonomicName id="4C7BADA3FF9BFF8D1045DE3CFE2BF9B6" box="[333,477,1551,1570]" class="Agaricomycetes" family="Polyporaceae" genus="Lentinus" kingdom="Fungi" order="Polyporales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="tigrinus">
|
||
<emphasis id="B90F0A32FF9BFF8D1045DE3CFE2BF9B6" bold="true" box="[333,477,1551,1570]" italics="true" pageId="2" pageNumber="3">Lentinus tigrinus</emphasis>
|
||
</taxonomicName>
|
||
,
|
||
<taxonomicName id="4C7BADA3FF9BFF8D10E0DE3CFD74F9B6" box="[488,642,1551,1570]" class="Agaricomycetes" family="Physalacriaceae" genus="Armillaria" kingdom="Fungi" order="Agaricales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="gallica">
|
||
<emphasis id="B90F0A32FF9BFF8D10E0DE3CFD74F9B6" bold="true" box="[488,642,1551,1570]" italics="true" pageId="2" pageNumber="3">Armillaria gallica</emphasis>
|
||
</taxonomicName>
|
||
,
|
||
<taxonomicName id="4C7BADA3FF9BFF8D1385DE3CFF2FF9AA" class="Tremellomycetes" family="Tremellaceae" genus="Cryptococcus" kingdom="Fungi" order="Tremellales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="depauperatus">
|
||
<emphasis id="B90F0A32FF9BFF8D1385DE3CFF2FF9AA" bold="true" italics="true" pageId="2" pageNumber="3">Cryptococcus depauperatus</emphasis>
|
||
</taxonomicName>
|
||
,
|
||
<emphasis id="B90F0A32FF9BFF8D11EEDE18FE5CF9AA" box="[230,426,1578,1598]" italics="true" pageId="2" pageNumber="3">
|
||
<emphasis id="B90F0A32FF9BFF8D11EEDE18FE8EF9AA" bold="true" box="[230,376,1579,1598]" italics="true" pageId="2" pageNumber="3">
|
||
<taxonomicName id="4C7BADA3FF9BFF8D11EEDE18FEBFF9AA" authorityName="Y.Yamada, H.Kawas., Itoh, I.Banno & Nakase" authorityYear="1988" box="[230,329,1579,1598]" class="Tremellomycetes" family="Tremellaceae" genus="Tsuchiyaea" kingdom="Fungi" order="Tremellales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="genus">Tsuchiyaea</taxonomicName>
|
||
wing
|
||
</emphasis>
|
||
fi
|
||
<emphasis id="B90F0A32FF9BFF8D108DDE18FE5CF9AA" bold="true" box="[389,426,1579,1598]" italics="true" pageId="2" pageNumber="3">eldii</emphasis>
|
||
</emphasis>
|
||
,
|
||
<taxonomicName id="4C7BADA3FF9BFF8D10BEDE18FDABF9AA" box="[438,605,1579,1598]" class="Tremellomycetes" family="Tremellaceae" genus="Cryptococcus" kingdom="Fungi" order="Tremellales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="gattii">
|
||
<emphasis id="B90F0A32FF9BFF8D10BEDE18FDABF9AA" bold="true" box="[438,605,1579,1598]" italics="true" pageId="2" pageNumber="3">Cryptococcus gattii</emphasis>
|
||
</taxonomicName>
|
||
and
|
||
<taxonomicName id="4C7BADA3FF9BFF8D1385DE18FF3DF9CD" class="Tremellomycetes" family="Tremellaceae" genus="Cryptococcus" kingdom="Fungi" order="Tremellales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="neoformans">
|
||
<emphasis id="B90F0A32FF9BFF8D1385DE18FF3DF9CD" bold="true" italics="true" pageId="2" pageNumber="3">Cryptococcus neoformans</emphasis>
|
||
</taxonomicName>
|
||
. LAG2 and LAG3 shared high identity with each other (displayed 66.06% identity), as well as to ceramide synthases (Cer2 and Cer3) of
|
||
<taxonomicName id="4C7BADA3FF9BFF8D11B6DE4DFEEFF905" box="[190,281,1662,1681]" class="Agaricomycetes" family="Polyporaceae" genus="Lentinus" kingdom="Fungi" order="Polyporales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="tigrinus">
|
||
<emphasis id="B90F0A32FF9BFF8D11B6DE4DFEEFF905" bold="true" box="[190,281,1662,1681]" italics="true" pageId="2" pageNumber="3">L. tigrinus</emphasis>
|
||
</taxonomicName>
|
||
(displayed 44.10% identity),
|
||
<taxonomicName id="4C7BADA3FF9BFF8D1348DE4DFD61F905" box="[576,663,1662,1681]" class="Agaricomycetes" family="Physalacriaceae" genus="Armillaria" kingdom="Fungi" order="Agaricales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="gallica">
|
||
<emphasis id="B90F0A32FF9BFF8D1348DE4DFD61F905" bold="true" box="[576,663,1662,1681]" italics="true" pageId="2" pageNumber="3">A. gallica</emphasis>
|
||
</taxonomicName>
|
||
(displayed 57.33% identity),
|
||
<taxonomicName id="4C7BADA3FF9BFF8D1028DEA9FE40F939" box="[288,438,1690,1709]" class="Tremellomycetes" family="Tremellaceae" genus="Cryptococcus" kingdom="Fungi" order="Tremellales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="depauperatus">
|
||
<emphasis id="B90F0A32FF9BFF8D1028DEA9FE40F939" bold="true" box="[288,438,1690,1709]" italics="true" pageId="2" pageNumber="3">C. depauperatus</emphasis>
|
||
</taxonomicName>
|
||
(displayed 49.88% identity),
|
||
<emphasis id="B90F0A32FF9BFF8D13E7DEA9FD0DF939" bold="true" box="[751,763,1690,1709]" italics="true" pageId="2" pageNumber="3">T</emphasis>
|
||
.
|
||
<emphasis id="B90F0A32FF9BFF8D116CDE85FF49F95D" box="[100,191,1718,1737]" italics="true" pageId="2" pageNumber="3">
|
||
<emphasis id="B90F0A32FF9BFF8D116CDE85FF7BF95D" bold="true" box="[100,141,1718,1737]" italics="true" pageId="2" pageNumber="3">wing</emphasis>
|
||
fi
|
||
<emphasis id="B90F0A32FF9BFF8D1192DE85FF49F95D" bold="true" box="[154,191,1718,1737]" italics="true" pageId="2" pageNumber="3">eldii</emphasis>
|
||
</emphasis>
|
||
(displayed 49.76% identity),
|
||
<taxonomicName id="4C7BADA3FF9BFF8D10FADE85FDC8F95D" box="[498,574,1718,1737]" class="Tremellomycetes" family="Tremellaceae" genus="Cryptococcus" kingdom="Fungi" order="Tremellales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="gattii">
|
||
<emphasis id="B90F0A32FF9BFF8D10FADE85FE09F95D" bold="true" box="[498,511,1718,1737]" italics="true" pageId="2" pageNumber="3">C</emphasis>
|
||
.
|
||
<emphasis id="B90F0A32FF9BFF8D131ADE85FDC8F95D" bold="true" box="[530,574,1718,1737]" italics="true" pageId="2" pageNumber="3">gattii</emphasis>
|
||
</taxonomicName>
|
||
(displayed 66.19% identity) and
|
||
<taxonomicName id="4C7BADA3FF9BFF8D11E6DEE1FE85F971" box="[238,371,1746,1765]" class="Tremellomycetes" family="Tremellaceae" genus="Cryptococcus" kingdom="Fungi" order="Tremellales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="neoformans">
|
||
<emphasis id="B90F0A32FF9BFF8D11E6DEE1FF0DF971" bold="true" box="[238,251,1746,1765]" italics="true" pageId="2" pageNumber="3">C</emphasis>
|
||
.
|
||
<emphasis id="B90F0A32FF9BFF8D1004DEE1FE85F971" bold="true" box="[268,371,1746,1765]" italics="true" pageId="2" pageNumber="3">neoformans</emphasis>
|
||
</taxonomicName>
|
||
(displayed 65.70% identity). The LAG1 amino acid sequence is greatly diverged from LAG2 and LAG3 (displayed 23.93% and 21.34% identity, respectively) in
|
||
<taxonomicName id="4C7BADA3FF9BFF8D1379DF39FD20F889" box="[625,726,1802,1821]" class="Agaricomycetes" family="Ganodermataceae" genus="Ganoderma" kingdom="Fungi" order="Polyporales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="lucidum">
|
||
<emphasis id="B90F0A32FF9BFF8D1379DF39FD20F889" bold="true" box="[625,726,1802,1821]" italics="true" pageId="2" pageNumber="3">G. lucidum</emphasis>
|
||
</taxonomicName>
|
||
and showed 19.14–36.98% identity to the proteins denoted as LAG1 or Cer
|
||
<quantity id="4C837BC5FF9BFF8D13F2DF15FF80F8C1" metricMagnitude="-2" metricUnit="m" metricValue="2.54" pageId="2" pageNumber="3" unit="in" value="1.0">1 in</quantity>
|
||
<taxonomicName id="4C7BADA3FF9BFF8D1175DF71FF20F8C1" box="[125,214,1858,1877]" class="Agaricomycetes" family="Polyporaceae" genus="Lentinus" kingdom="Fungi" order="Polyporales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="tigrinus">
|
||
<emphasis id="B90F0A32FF9BFF8D1175DF71FF7EF8C1" bold="true" box="[125,136,1858,1877]" italics="true" pageId="2" pageNumber="3">L</emphasis>
|
||
.
|
||
<emphasis id="B90F0A32FF9BFF8D119DDF71FF20F8C1" bold="true" box="[149,214,1858,1877]" italics="true" pageId="2" pageNumber="3">tigrinus</emphasis>
|
||
</taxonomicName>
|
||
(36.98%),
|
||
<taxonomicName id="4C7BADA3FF9BFF8D104ADF71FE61F8C1" box="[322,407,1858,1877]" class="Agaricomycetes" family="Physalacriaceae" genus="Armillaria" kingdom="Fungi" order="Agaricales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="gallica">
|
||
<emphasis id="B90F0A32FF9BFF8D104ADF71FEA6F8C1" bold="true" box="[322,336,1858,1877]" italics="true" pageId="2" pageNumber="3">A</emphasis>
|
||
.
|
||
<emphasis id="B90F0A32FF9BFF8D1055DF71FE61F8C1" bold="true" box="[349,407,1858,1877]" italics="true" pageId="2" pageNumber="3">gallica</emphasis>
|
||
</taxonomicName>
|
||
(26.88%),
|
||
<taxonomicName id="4C7BADA3FF9BFF8D130ADF71FD75F8C1" box="[514,643,1858,1877]" class="Tremellomycetes" family="Tremellaceae" genus="Cryptococcus" kingdom="Fungi" order="Tremellales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="neoformans">
|
||
<emphasis id="B90F0A32FF9BFF8D130ADF71FDF9F8C1" bold="true" box="[514,527,1858,1877]" italics="true" pageId="2" pageNumber="3">C</emphasis>
|
||
.
|
||
<emphasis id="B90F0A32FF9BFF8D1314DF71FD75F8C1" bold="true" box="[540,643,1858,1877]" italics="true" pageId="2" pageNumber="3">neoformans</emphasis>
|
||
</taxonomicName>
|
||
(35.84%),
|
||
<taxonomicName id="4C7BADA3FF9BFF8D13E6DF71FF2FF8E4" class="Tremellomycetes" family="Tremellaceae" genus="Cryptococcus" kingdom="Fungi" order="Tremellales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="depauperatus">
|
||
<emphasis id="B90F0A32FF9BFF8D13E6DF71FD0DF8C1" bold="true" box="[750,763,1858,1877]" italics="true" pageId="2" pageNumber="3">C</emphasis>
|
||
.
|
||
<emphasis id="B90F0A32FF9BFF8D116CDF6EFF2FF8E4" bold="true" box="[100,217,1885,1904]" italics="true" pageId="2" pageNumber="3">depauperatus</emphasis>
|
||
</taxonomicName>
|
||
(33.08%),
|
||
<emphasis id="B90F0A32FF9BFF8D1041DF6EFEA3F8E4" bold="true" box="[329,341,1885,1904]" italics="true" pageId="2" pageNumber="3">T</emphasis>
|
||
.
|
||
<emphasis id="B90F0A32FF9BFF8D106CDF6EFE48F8E4" box="[356,446,1885,1904]" italics="true" pageId="2" pageNumber="3">
|
||
<emphasis id="B90F0A32FF9BFF8D106CDF6EFE7BF8E4" bold="true" box="[356,397,1885,1904]" italics="true" pageId="2" pageNumber="3">wing</emphasis>
|
||
fi
|
||
<emphasis id="B90F0A32FF9BFF8D1091DF6EFE48F8E4" bold="true" box="[409,446,1885,1904]" italics="true" pageId="2" pageNumber="3">eldii</emphasis>
|
||
</emphasis>
|
||
(32.59%) and
|
||
<taxonomicName id="4C7BADA3FF9BFF8D135BDF6EFD6DF8E4" box="[595,667,1885,1904]" class="Tremellomycetes" family="Tremellaceae" genus="Cryptococcus" kingdom="Fungi" order="Tremellales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="gattii">
|
||
<emphasis id="B90F0A32FF9BFF8D135BDF6EFD96F8E4" bold="true" box="[595,608,1885,1904]" italics="true" pageId="2" pageNumber="3">C</emphasis>
|
||
.
|
||
<emphasis id="B90F0A32FF9BFF8D1367DF6EFD6DF8E4" bold="true" box="[623,667,1885,1904]" italics="true" pageId="2" pageNumber="3">gattii</emphasis>
|
||
</taxonomicName>
|
||
(36.90%), Lac
|
||
<quantity id="4C837BC5FF9BFF8D1181DF4AFF59F818" box="[137,175,1913,1932]" metricMagnitude="-2" metricUnit="m" metricValue="2.54" pageId="2" pageNumber="3" unit="in" value="1.0">1 in</quantity>
|
||
<taxonomicName id="4C7BADA3FF9BFF8D11B4DF4AFE99F818" box="[188,367,1913,1932]" class="Tremellomycetes" family="Tremellaceae" genus="Blastomyces" kingdom="Fungi" order="Tremellales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="parvus">
|
||
<emphasis id="B90F0A32FF9BFF8D11B4DF4AFE99F818" bold="true" box="[188,367,1913,1932]" italics="true" pageId="2" pageNumber="3">Blastomyces parvus</emphasis>
|
||
</taxonomicName>
|
||
(20.11%),
|
||
<taxonomicName id="4C7BADA3FF9BFF8D10EFDF4AFD61F818" box="[487,663,1913,1932]" class="Agaricomycetes" family="Fomitopsidaceae" genus="Emmonsia" kingdom="Fungi" order="Polyporales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="crescens">
|
||
<emphasis id="B90F0A32FF9BFF8D10EFDF4AFD61F818" bold="true" box="[487,663,1913,1932]" italics="true" pageId="2" pageNumber="3">Emmonsia crescens</emphasis>
|
||
</taxonomicName>
|
||
(19.14%),
|
||
<taxonomicName id="4C7BADA3FF9BFF8D116CDFA6FEE6F83C" box="[100,272,1941,1960]" class="Saccharomycetes" family="Dipodascaceae" genus="Yarrowia" kingdom="Fungi" order="Saccharomycetales" pageId="2" pageNumber="3" phylum="Ascomycota" rank="species" species="lipolytica">
|
||
<emphasis id="B90F0A32FF9BFF8D116CDFA6FEE6F83C" bold="true" box="[100,272,1941,1960]" italics="true" pageId="2" pageNumber="3">Yarrowia lipolytica</emphasis>
|
||
</taxonomicName>
|
||
(30.26%),
|
||
<taxonomicName id="4C7BADA3FF9BFF8D1075DFA6FD92F83C" box="[381,612,1941,1960]" class="Saccharomycetes" family="Wickerhamomycetaceae" genus="Wickerhamomyces" kingdom="Fungi" order="Saccharomycetales" pageId="2" pageNumber="3" phylum="Ascomycota" rank="species" species="ciferrii">
|
||
<emphasis id="B90F0A32FF9BFF8D1075DFA6FD92F83C" bold="true" box="[381,612,1941,1960]" italics="true" pageId="2" pageNumber="3">Wickerhamomyces ciferrii</emphasis>
|
||
</taxonomicName>
|
||
(26.12%),
|
||
<taxonomicName id="4C7BADA3FF9BFF8D13DADFA6FEC4F850" class="Saccharomycetes" family="Saccharomycetaceae" genus="Kluyveromyces" kingdom="Fungi" order="Saccharomycetales" pageId="2" pageNumber="3" phylum="Ascomycota" rank="species" species="marxianus">
|
||
<emphasis id="B90F0A32FF9BFF8D13DADFA6FEC4F850" bold="true" italics="true" pageId="2" pageNumber="3">Kluyveromyces marxianus</emphasis>
|
||
</taxonomicName>
|
||
(27.98%),
|
||
<taxonomicName id="4C7BADA3FF9BFF8D10BFDF82FDA8F850" box="[439,606,1969,1988]" class="Tremellomycetes" family="Tremellaceae" genus="Candida" kingdom="Fungi" order="Tremellales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="albicans">
|
||
<emphasis id="B90F0A32FF9BFF8D10BFDF82FDA8F850" bold="true" box="[439,606,1969,1988]" italics="true" pageId="2" pageNumber="3">Candida albicans</emphasis>
|
||
</taxonomicName>
|
||
(24.25%) and
|
||
<taxonomicName id="4C7BADA3FF9BFF8D123AD8ACFC1DFF26" box="[818,1003,159,178]" class="Saccharomycetes" family="Saccharomycetaceae" genus="Kluyveromyces" kingdom="Fungi" order="Saccharomycetales" pageId="2" pageNumber="3" phylum="Ascomycota" rank="species" species="lactis">
|
||
<emphasis id="B90F0A32FF9BFF8D123AD8ACFC1DFF26" bold="true" box="[818,1003,159,178]" italics="true" pageId="2" pageNumber="3">Kluyveromyces lactis</emphasis>
|
||
</taxonomicName>
|
||
(32.29%), and Bar1p in
|
||
<taxonomicName id="4C7BADA3FF9BFF8D15D2D8ACFAADFF26" box="[1242,1371,159,178]" class="Saccharomycetes" family="Pichiaceae" genus="Pichia" kingdom="Fungi" order="Saccharomycetales" pageId="2" pageNumber="3" phylum="Ascomycota" rank="species" species="pastoris">
|
||
<emphasis id="B90F0A32FF9BFF8D15D2D8ACFAADFF26" bold="true" box="[1242,1371,159,178]" italics="true" pageId="2" pageNumber="3">Pichia pastoris</emphasis>
|
||
</taxonomicName>
|
||
(27.98%).
|
||
</paragraph>
|
||
<paragraph id="8BC4D620FF9BFF8D125BD888FCA0FE06" blockId="2.[818,1488,159,402]" pageId="2" pageNumber="3">
|
||
The conserved domain analysis for the CERS proteins was performed using the SMART database (http://smart.emblheidelberg.de). The domain analysis revealed that the three predicted
|
||
<taxonomicName id="4C7BADA3FF9BFF8D1426D8C0FA79FE92" box="[1326,1423,243,262]" class="Agaricomycetes" family="Ganodermataceae" genus="Ganoderma" kingdom="Fungi" order="Polyporales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="lucidum">
|
||
<emphasis id="B90F0A32FF9BFF8D1426D8C0FA79FE92" bold="true" box="[1326,1423,243,262]" italics="true" pageId="2" pageNumber="3">G. lucidum</emphasis>
|
||
</taxonomicName>
|
||
CERSs contain one Tram-Lag-CLN8 (TLC) domain. The TLC domains of LAG2 and LAG3 are highly similar to each other and are different from that of LAG1. In addition, LAG2 and LAG3 have two transmembrane (TM) domains, whereas LAG1 has only one TM domain (Supplementary
|
||
<figureCitation id="1340CAA5FF9BFF8D14A4D950FCBFFE06" captionStart="Fig" captionStartId="1.[1116,1146,152,169]" captionTargetBox="[103,1084,155,1016]" captionTargetId="figure-665@1.[100,1086,151,1018]" captionTargetPageId="1" captionText="Fig. 1. Overview of the sphingolipid biosynthetic pathway in yeast and filamentous fungi. The abbreviations used are as follows: Dihydroxy LCB: dihydroxy longchain (sphingoid) base; Trihydroxy LCB: trihydroxy long-chain sphingoid base; Long chain FA-CoA: long chain fatty acyl-coenzyme A; Very long chain FA-CoA: very long chain fatty acyl-coenzyme A; IPC synthase: inositol phosphorylceramide synthase; IPC mannosyl transferase: inositol phosphorylceramide mannosyl transferase; MIPCs: mannose inositol phosphorylceramides; M (IP)2Cs: mannose (inositol phosphoryl)2- ceramides." figureDoi="http://doi.org/10.5281/zenodo.8294208" httpUri="https://zenodo.org/record/8294208/files/figure.png" pageId="2" pageNumber="3">Fig. S1</figureCitation>
|
||
).
|
||
</paragraph>
|
||
<paragraph id="8BC4D620FF9BFF8D123AD985FAF6FE5D" blockId="2.[818,1280,438,457]" box="[818,1280,438,457]" pageId="2" pageNumber="3">
|
||
<heading id="D08C614CFF9BFF8D123AD985FAF6FE5D" bold="true" box="[818,1280,438,457]" fontSize="36" level="1" pageId="2" pageNumber="3" reason="1">
|
||
<emphasis id="B90F0A32FF9BFF8D123AD985FAF6FE5D" bold="true" box="[818,1280,438,457]" italics="true" pageId="2" pageNumber="3">
|
||
2.2. Construction of
|
||
<taxonomicName id="4C7BADA3FF9BFF8D12F8D985FB49FE5D" box="[1008,1215,438,457]" class="Agaricomycetes" family="Ganodermataceae" genus="Ganoderma" kingdom="Fungi" order="Polyporales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="subSpecies" species="lucidum" subSpecies="lag-silenced">G. lucidum lag-silenced</taxonomicName>
|
||
strains
|
||
</emphasis>
|
||
</heading>
|
||
</paragraph>
|
||
<paragraph id="8BC4D620FF9BFF8D125BD9DDFA53FCA0" blockId="2.[818,1487,494,820]" pageId="2" pageNumber="3">
|
||
To investigate the function of CERS in physiological processes, we constructed
|
||
<emphasis id="B90F0A32FF9BFF8D12A0DA39FC3BFD89" bold="true" box="[936,973,522,541]" italics="true" pageId="2" pageNumber="3">lag1</emphasis>
|
||
,
|
||
<emphasis id="B90F0A32FF9BFF8D12D4DA39FBF4FD89" bold="true" box="[988,1026,522,541]" italics="true" pageId="2" pageNumber="3">lag2</emphasis>
|
||
and
|
||
<emphasis id="B90F0A32FF9BFF8D153FDA39FBABFD89" bold="true" box="[1079,1117,522,541]" italics="true" pageId="2" pageNumber="3">lag3</emphasis>
|
||
single-silenced mutants and
|
||
<emphasis id="B90F0A32FF9BFF8D1471DA39FA39FD89" bold="true" box="[1401,1487,522,541]" italics="true" pageId="2" pageNumber="3">lag2/lag3</emphasis>
|
||
double-silenced mutants as previously described by using an established dual-promoter system (Supplementary
|
||
<figureCitation id="1340CAA5FF9BFF8D15D4DA71FAEDFDC1" box="[1244,1307,578,597]" captionStart="Fig" captionStartId="2.[100,130,1274,1291]" captionTargetBox="[106,765,154,1243]" captionTargetId="figure-844@2.[105,766,152,1251]" captionTargetPageId="2" captionText="Fig. 2. G. lucidum harbours three ceramide synthases. Phylogenetic analysis of ceramide synthases in eukaryotes. The phylogenetic tree was conducted using MEGA 6 and can be roughly divided into three major groups: human, plant and fungi. The evolutionary history was inferred by the neighbour-joining method from 1000 replicates using MEGA 6." figureDoi="http://doi.org/10.5281/zenodo.8294210" httpUri="https://zenodo.org/record/8294210/files/figure.png" pageId="2" pageNumber="3">Fig. S2</figureCitation>
|
||
), which carries the hygromycin B (Hyg) resistance gene as a selectable marker. Real-time PCR analysis was used to examine the expression of the
|
||
<emphasis id="B90F0A32FF9BFF8D1447DA49FA9FFD19" bold="true" box="[1359,1385,634,653]" italics="true" pageId="2" pageNumber="3">lag</emphasis>
|
||
genes and confirm silencing in the transformants. The results showed that the expression of
|
||
<emphasis id="B90F0A32FF9BFF8D12BCDA82FC38FD50" bold="true" box="[948,974,689,708]" italics="true" pageId="2" pageNumber="3">lag</emphasis>
|
||
was down-regulated greater than 50% (
|
||
<emphasis id="B90F0A32FF9BFF8D1445DA82FAAFFD50" bold="true" box="[1357,1369,689,708]" italics="true" pageId="2" pageNumber="3">P</emphasis>
|
||
<0.01) in all
|
||
<emphasis id="B90F0A32FF9BFF8D125BDAFEFC9BFD74" bold="true" box="[851,877,717,736]" italics="true" pageId="2" pageNumber="3">lag</emphasis>
|
||
-silenced strains compared to that in WT strains. The silenced strains
|
||
<emphasis id="B90F0A32FF9BFF8D127DDADAFC57FD68" bold="true" box="[885,929,745,764]" italics="true" pageId="2" pageNumber="3">lag1-</emphasis>
|
||
(LAG1i-7 and LAG1i-8),
|
||
<emphasis id="B90F0A32FF9BFF8D1584DADAFB4EFD68" bold="true" box="[1164,1208,745,764]" italics="true" pageId="2" pageNumber="3">lag2-</emphasis>
|
||
(LAG2i-1 and LAG2i-7),
|
||
<emphasis id="B90F0A32FF9BFF8D14AADADAFA39FD68" bold="true" box="[1442,1487,745,764]" italics="true" pageId="2" pageNumber="3">lag3-</emphasis>
|
||
(LAG3i-22 and LAG3i-26) and
|
||
<emphasis id="B90F0A32FF9BFF8D1561DB36FB30FC8C" bold="true" box="[1129,1222,773,792]" italics="true" pageId="2" pageNumber="3">lag2/lag3-</emphasis>
|
||
(LAG23i-4 and LAG23i-5) strains were selected for further analysis (Supplementary
|
||
<figureCitation id="1340CAA5FF9BFF8D145FDB12FA6EFCA0" box="[1367,1432,801,820]" captionStart="Fig" captionStartId="3.[1116,1146,152,169]" captionTargetBox="[107,1064,173,658]" captionTargetId="figure-350@3.[100,1086,151,678]" captionTargetPageId="3" captionText="Fig. 3. Effect of lag gene silencing on G. lucidum growth. Morphology of fungal colonies in lag-silenced, WT and SiControl strains after cultivation in the dark on CYM medium at 28 ̊C for 5 days. Three independent biological replicates in each column. Error bars represent standard deviations, and asterisks show significant differences from control (WT and Sicontrol) strains according to Student's t-test (**P <0.01, n = 3)." figureDoi="http://doi.org/10.5281/zenodo.8294212" httpUri="https://zenodo.org/record/8294212/files/figure.png" pageId="2" pageNumber="3">Fig. S3</figureCitation>
|
||
).
|
||
</paragraph>
|
||
<paragraph id="8BC4D620FF9BFF8D123ADB6AFC98FC1C" blockId="2.[818,1451,857,904]" pageId="2" pageNumber="3">
|
||
<heading id="D08C614CFF9BFF8D123ADB6AFC98FC1C" bold="true" fontSize="36" level="1" pageId="2" pageNumber="3" reason="1">
|
||
<emphasis id="B90F0A32FF9BFF8D123ADB6AFC98FC1C" bold="true" italics="true" pageId="2" pageNumber="3">
|
||
2.3. Ceramide synthases
|
||
<collectionCode id="ED6A4EE5FF9BFF8D151DDB6AFBC9FCF8" box="[1045,1087,857,876]" pageId="2" pageNumber="3">LAG</collectionCode>
|
||
2 and
|
||
<collectionCode id="ED6A4EE5FF9BFF8D1571DB6AFB55FCF8" box="[1145,1187,857,876]" pageId="2" pageNumber="3">LAG</collectionCode>
|
||
3 are required for
|
||
<taxonomicName id="4C7BADA3FF9BFF8D1440DB6AFA5DFCF8" box="[1352,1451,857,876]" class="Agaricomycetes" family="Ganodermataceae" genus="Ganoderma" kingdom="Fungi" order="Polyporales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="subSpecies" species="lucidum" subSpecies="growth">G. lucidum</taxonomicName>
|
||
growth
|
||
</emphasis>
|
||
</heading>
|
||
</paragraph>
|
||
<paragraph id="8BC4D620FF9BFF8D125BDB9FFB0DFA46" blockId="2.[818,1488,940,1490]" pageId="2" pageNumber="3">
|
||
To explore whether
|
||
<collectionCode id="ED6A4EE5FF9BFF8D1516DB9FFBA7FC2B" box="[1054,1105,940,959]" pageId="2" pageNumber="3">CERS</collectionCode>
|
||
impact the growth of
|
||
<taxonomicName id="4C7BADA3FF9BFF8D1433DB9FFA57FC2B" box="[1339,1441,940,959]" class="Agaricomycetes" family="Ganodermataceae" genus="Ganoderma" kingdom="Fungi" order="Polyporales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="species" species="lucidum">
|
||
<emphasis id="B90F0A32FF9BFF8D1433DB9FFA57FC2B" bold="true" box="[1339,1441,940,959]" italics="true" pageId="2" pageNumber="3">G. lucidum</emphasis>
|
||
</taxonomicName>
|
||
, the SiControl and WT strains and the individual
|
||
<emphasis id="B90F0A32FF9BFF8D15FFDBFBFAE7FC4F" bold="true" box="[1271,1297,968,987]" italics="true" pageId="2" pageNumber="3">lag</emphasis>
|
||
- and the
|
||
<emphasis id="B90F0A32FF9BFF8D1471DBFBFA39FC4F" bold="true" box="[1401,1487,968,987]" italics="true" pageId="2" pageNumber="3">lag2/lag3</emphasis>
|
||
double-silenced transformants were cultivated on solid CYM for 5 days to evaluate the resulting colony diameters. The colony diameters of the individual
|
||
<emphasis id="B90F0A32FF9BFF8D12AADC2FFC4AFBBB" bold="true" box="[930,956,1052,1071]" italics="true" pageId="2" pageNumber="3">lag</emphasis>
|
||
-silenced transformants
|
||
<collectionCode id="ED6A4EE5FF9BFF8D15BBDC2FFB12FBBB" box="[1203,1252,1052,1071]" pageId="2" pageNumber="3">LAG</collectionCode>
|
||
1i-7,
|
||
<collectionCode id="ED6A4EE5FF9BFF8D141FDC2FFABFFBBB" box="[1303,1353,1052,1071]" pageId="2" pageNumber="3">LAG</collectionCode>
|
||
1i-8,
|
||
<collectionCode id="ED6A4EE5FF9BFF8D1474DC2FFA5BFBBB" box="[1404,1453,1052,1071]" pageId="2" pageNumber="3">LAG</collectionCode>
|
||
2i-1,
|
||
<collectionCode id="ED6A4EE5FF9BFF8D123ADC0BFC97FBDF" box="[818,865,1080,1099]" pageId="2" pageNumber="3">LAG</collectionCode>
|
||
2i-7,
|
||
<collectionCode id="ED6A4EE5FF9BFF8D129BDC0BFC37FBDF" box="[915,961,1080,1099]" pageId="2" pageNumber="3">LAG</collectionCode>
|
||
3i-22 and
|
||
<collectionCode id="ED6A4EE5FF9BFF8D1524DC0BFBACFBDF" box="[1068,1114,1080,1099]" pageId="2" pageNumber="3">LAG</collectionCode>
|
||
3i-26 were not significantly different (
|
||
<emphasis id="B90F0A32FF9BFF8D1232DC67FCB0FBF3" bold="true" box="[826,838,1108,1127]" italics="true" pageId="2" pageNumber="3">
|
||
<collectionCode id="ED6A4EE5FF9BFF8D1232DC67FCB0FBF3" box="[826,838,1108,1127]" country="France" lsid="urn:lsid:biocol.org:col:15763" name="Museum National d' Histoire Naturelle, Paris (MNHN) - Vascular Plants" pageId="2" pageNumber="3" type="Herbarium">P</collectionCode>
|
||
</emphasis>
|
||
> 0.05) from those of the WT and SiControl strains (
|
||
<figureCitation id="1340CAA5FF9BFF8D1463DC67FA55FBF3" box="[1387,1443,1108,1127]" captionStart="Fig" captionStartId="3.[1116,1146,152,169]" captionTargetBox="[107,1064,173,658]" captionTargetId="figure-350@3.[100,1086,151,678]" captionTargetPageId="3" captionText="Fig. 3. Effect of lag gene silencing on G. lucidum growth. Morphology of fungal colonies in lag-silenced, WT and SiControl strains after cultivation in the dark on CYM medium at 28 ̊C for 5 days. Three independent biological replicates in each column. Error bars represent standard deviations, and asterisks show significant differences from control (WT and Sicontrol) strains according to Student's t-test (**P <0.01, n = 3)." figureDoi="http://doi.org/10.5281/zenodo.8294212" httpUri="https://zenodo.org/record/8294212/files/figure.png" pageId="2" pageNumber="3">Fig. 3</figureCitation>
|
||
and Supplementary
|
||
<figureCitation id="1340CAA5FF9BFF8D12C1DC43FBECFB17" box="[969,1050,1136,1155]" captionStart="Fig" captionStartId="3.[100,130,1889,1906]" captionTargetBox="[264,1323,1260,1866]" captionTargetId="figure-431@3.[263,1325,1258,1867]" captionTargetPageId="3" captionText="Fig. 4. Ganoderic acid biosynthesis in G. lucidum is influenced by the lag gene. A Systematic content analysis of ganoderic acid in the lag-silenced, WT and SiControl strains. B–D Relative gene expression of hmgr (B), sqs (C) and osc (D) in the WT, SiControl and lag-silenced strains. There are three independent biological replicates in each column. Error bars represent standard deviations, and asterisks show significant differences from control (WT and Sicontrol) strains according to Student's t-test (**P <0.01, n = 3)." figureDoi="http://doi.org/10.5281/zenodo.8294214" httpUri="https://zenodo.org/record/8294214/files/figure.png" pageId="2" pageNumber="3">Fig. S4A</figureCitation>
|
||
). For the l
|
||
<emphasis id="B90F0A32FF9BFF8D1580DC43FB5EFB17" bold="true" box="[1160,1192,1136,1155]" italics="true" pageId="2" pageNumber="3">ag2</emphasis>
|
||
/
|
||
<emphasis id="B90F0A32FF9BFF8D15BBDC43FB2FFB17" bold="true" box="[1203,1241,1136,1155]" italics="true" pageId="2" pageNumber="3">lag3</emphasis>
|
||
double-silenced transformants
|
||
<collectionCode id="ED6A4EE5FF9BFF8D127EDCBFFC52FB0B" box="[886,932,1164,1183]" pageId="2" pageNumber="3">LAG</collectionCode>
|
||
23i-4 and
|
||
<collectionCode id="ED6A4EE5FF9BFF8D1503DCBFFBCDFB0B" box="[1035,1083,1164,1183]" pageId="2" pageNumber="3">LAG</collectionCode>
|
||
23i-5, growth was decreased by approximately 22% and 45% (
|
||
<emphasis id="B90F0A32FF9BFF8D151DDC9BFBD7FB2F" bold="true" box="[1045,1057,1192,1211]" italics="true" pageId="2" pageNumber="3">
|
||
<collectionCode id="ED6A4EE5FF9BFF8D151DDC9BFBD7FB2F" box="[1045,1057,1192,1211]" country="France" lsid="urn:lsid:biocol.org:col:15763" name="Museum National d' Histoire Naturelle, Paris (MNHN) - Vascular Plants" pageId="2" pageNumber="3" type="Herbarium">P</collectionCode>
|
||
</emphasis>
|
||
<0.01), respectively, compared with that observed for the SiControl and WT strains. We further evaluated the effects of the loss of the
|
||
<emphasis id="B90F0A32FF9BFF8D1519DCECFBDDFB66" bold="true" box="[1041,1067,1247,1266]" italics="true" pageId="2" pageNumber="3">lag</emphasis>
|
||
genes on
|
||
<taxonomicName id="4C7BADA3FF9BFF8D1580DCECFAC7FB66" box="[1160,1329,1247,1266]" class="Agaricomycetes" family="Ganodermataceae" genus="Ganoderma" kingdom="Fungi" order="Polyporales" pageId="2" pageNumber="3" phylum="Basidiomycota" rank="subSpecies" species="lucidum" subSpecies="growth">
|
||
<emphasis id="B90F0A32FF9BFF8D1580DCECFB1FFB66" bold="true" box="[1160,1257,1247,1266]" italics="true" pageId="2" pageNumber="3">G. lucidum</emphasis>
|
||
growth
|
||
</taxonomicName>
|
||
by analysing the biomass of the strains. Compared to the SiControl and WT strains, the dry weights of the individual
|
||
<emphasis id="B90F0A32FF9BFF8D155ADD24FB9AFABE" bold="true" box="[1106,1132,1303,1322]" italics="true" pageId="2" pageNumber="3">lag</emphasis>
|
||
-silenced transformants were not significantly different (
|
||
<emphasis id="B90F0A32FF9BFF8D12FFDD00FBF5FAD2" bold="true" box="[1015,1027,1331,1350]" italics="true" pageId="2" pageNumber="3">
|
||
<collectionCode id="ED6A4EE5FF9BFF8D12FFDD00FBF5FAD2" box="[1015,1027,1331,1350]" country="France" lsid="urn:lsid:biocol.org:col:15763" name="Museum National d' Histoire Naturelle, Paris (MNHN) - Vascular Plants" pageId="2" pageNumber="3" type="Herbarium">P</collectionCode>
|
||
</emphasis>
|
||
> 0.05); however, the dry weights were significantly (
|
||
<emphasis id="B90F0A32FF9BFF8D1295DD7CFC5FFAF6" bold="true" box="[925,937,1359,1378]" italics="true" pageId="2" pageNumber="3">
|
||
<collectionCode id="ED6A4EE5FF9BFF8D1295DD7CFC5FFAF6" box="[925,937,1359,1378]" country="France" lsid="urn:lsid:biocol.org:col:15763" name="Museum National d' Histoire Naturelle, Paris (MNHN) - Vascular Plants" pageId="2" pageNumber="3" type="Herbarium">P</collectionCode>
|
||
</emphasis>
|
||
<0.01) decreased by approximately 44 and 49%, respectively, in the
|
||
<collectionCode id="ED6A4EE5FF9BFF8D12E8DD58FBF8FAEA" box="[992,1038,1387,1406]" pageId="2" pageNumber="3">LAG</collectionCode>
|
||
23i-4 and
|
||
<collectionCode id="ED6A4EE5FF9BFF8D1567DD58FB68FAEA" box="[1135,1182,1387,1406]" pageId="2" pageNumber="3">LAG</collectionCode>
|
||
23i-5 strains compared with the SiControl and WT strains (Supplementary
|
||
<figureCitation id="1340CAA5FF9BFF8D15C3DDB4FAEDFA0E" box="[1227,1307,1415,1434]" captionStart="Fig" captionStartId="3.[100,130,1889,1906]" captionTargetBox="[264,1323,1260,1866]" captionTargetId="figure-431@3.[263,1325,1258,1867]" captionTargetPageId="3" captionText="Fig. 4. Ganoderic acid biosynthesis in G. lucidum is influenced by the lag gene. A Systematic content analysis of ganoderic acid in the lag-silenced, WT and SiControl strains. B–D Relative gene expression of hmgr (B), sqs (C) and osc (D) in the WT, SiControl and lag-silenced strains. There are three independent biological replicates in each column. Error bars represent standard deviations, and asterisks show significant differences from control (WT and Sicontrol) strains according to Student's t-test (**P <0.01, n = 3)." figureDoi="http://doi.org/10.5281/zenodo.8294214" httpUri="https://zenodo.org/record/8294214/files/figure.png" pageId="2" pageNumber="3">Fig. S4B</figureCitation>
|
||
). These results indicated that
|
||
<emphasis id="B90F0A32FF9BFF8D12A2DD90FC39FA22" bold="true" box="[938,975,1443,1462]" italics="true" pageId="2" pageNumber="3">lag2</emphasis>
|
||
and
|
||
<emphasis id="B90F0A32FF9BFF8D1508DD90FBD0FA22" bold="true" box="[1024,1062,1443,1462]" italics="true" pageId="2" pageNumber="3">lag3</emphasis>
|
||
are required and are functionally redundant with respect to the observed growth phenotype.
|
||
</paragraph>
|
||
</subSubSection>
|
||
</treatment>
|
||
</document> |