215 lines
29 KiB
XML
215 lines
29 KiB
XML
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<mods:title id="74C60B4D9DEF8AA97809C1C8FF4F4ACC">Influences of hydrozoan colonization on proteomic profiles of the brown alga Saccharina japonica</mods:title>
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<mods:namePart id="A15E891BE6CBF896D26E71BBB6669EB2">Getachew, Paulos</mods:namePart>
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<mods:namePart id="502C9EB56268AC98F7416F7969B9C58E">Nam, Bo-Hye</mods:namePart>
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<mods:namePart id="BECC54F43D55B90E39DEB3BBBD0711C8">Young Cho, Ji</mods:namePart>
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<mods:namePart id="69FFFAB595D7940EB34EAE9E92165148">Hong, Yong-Ki</mods:namePart>
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<mods:title id="1A7912DD9EC4CA3511ACD3CA3F369B6B">Botanica Marina</mods:title>
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<mods:title id="1EA96B59375E82786495DF918AB4FFF1">Warsaw, Poland</mods:title>
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<treatment id="03EA879AFFBDFFCFFF2BFB85FE2CFD81" ID-DOI="http://doi.org/10.5281/zenodo.11493311" ID-GBIF-Taxon="232142848" ID-Zenodo-Dep="11493311" LSID="urn:lsid:plazi:treatment:03EA879AFFBDFFCFFF2BFB85FE2CFD81" httpUri="http://treatment.plazi.org/id/03EA879AFFBDFFCFFF2BFB85FE2CFD81" lastPageId="5" lastPageNumber="87" pageId="1" pageNumber="86">
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<subSubSection id="C3596507FFBDFFCBFF2BFB85FEDCFB3C" pageId="1" pageNumber="86" type="nomenclature">
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<paragraph id="8BFC368CFFBDFFCBFF2BFB85FEDCFB3C" blockId="1.[166,681,1101,1171]" pageId="1" pageNumber="86">
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<heading id="D0B481E0FFBDFFCBFF2BFB85FEDCFB3C" fontSize="12" level="2" pageId="1" pageNumber="86" reason="7">
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<taxonomicName id="4C434D0FFFBDFFCBFF2BFB85FE3DFBC4" baseAuthorityName="Park and Hwang" baseAuthorityYear="2012" box="[166,449,1101,1132]" class="Phaeophyceae" family="Laminariaceae" genus="Saccharina" kingdom="Chromista" order="Laminariales" pageId="1" pageNumber="86" phylum="Ochrophyta" rank="species" species="japonica">Saccharina japonica</taxonomicName>
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, hydrozoan, and reagents
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</heading>
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</paragraph>
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</subSubSection>
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<subSubSection id="C3596507FFBDFFCBFF2BFB72FDB7F918" pageId="1" pageNumber="86" type="materials_examined">
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<paragraph id="8BFC368CFFBDFFCBFF2BFB72FDB7F918" blockId="1.[166,794,1208,1719]" pageId="1" pageNumber="86">
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Fresh blades of late-harvested
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<taxonomicName id="4C434D0FFFBDFFCBFE73FB70FD21FB7D" baseAuthorityName="Park and Hwang" baseAuthorityYear="2012" box="[510,733,1208,1234]" class="Phaeophyceae" family="Laminariaceae" genus="Saccharina" kingdom="Chromista" order="Laminariales" pageId="1" pageNumber="86" phylum="Ochrophyta" rank="species" species="japonica">
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<emphasis id="B937EA9EFFBDFFCBFE73FB70FD21FB7D" bold="true" box="[510,733,1208,1234]" italics="true" pageId="1" pageNumber="86">Saccharina japonica</emphasis>
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</taxonomicName>
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were collected from the Gijang aquaculture farm,
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<collectingRegion id="4987F86EFFBDFFCBFD04FB15FD2DFB5B" box="[649,721,1245,1268]" country="South Korea" name="Busan" pageId="1" pageNumber="86">Busan</collectingRegion>
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,
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<collectingCountry id="F354761CFFBDFFCBFD56FB15FCE6FB5B" box="[731,794,1245,1268]" name="South Korea" pageId="1" pageNumber="86">Korea</collectingCountry>
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in
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<date id="FFFD104CFFBDFFCBFF4BFAC8FECDFAB8" box="[198,305,1279,1303]" pageId="1" pageNumber="86" value="2013-06">
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<collectingDate id="EFB9E9A4FFBDFFCBFF4BFAC8FECDFAB8" box="[198,305,1279,1303]" pageId="1" pageNumber="86" value="2013-06">June 2013</collectingDate>
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</date>
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and 2014. A voucher specimen was deposited in the author’s laboratory (
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<collectorName id="26B6535AFFBDFFCBFD8CFAEAFD86FA95" box="[513,634,1314,1338]" pageId="1" pageNumber="86">Y. K. Hong</collectorName>
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||
).
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<collectorName id="26B6535AFFBDFFCBFD00FAEAFD4AFA95" box="[653,694,1314,1338]" pageId="1" pageNumber="86">The</collectorName>
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seaweed tissues were washed and cleaned with autoclaved seawater.
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<collectorName id="26B6535AFFBDFFCBFF5DFAAFFED2FAD0" box="[208,302,1383,1407]" pageId="1" pageNumber="86">Colonies</collectorName>
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of
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<taxonomicName id="4C434D0FFFBDFFCBFEDDFAAEFE21FAD0" box="[336,477,1382,1408]" class="Hydrozoa" family="Campanulariidae" genus="Obelia" kingdom="Animalia" order="Leptothecata" pageId="1" pageNumber="86" phylum="Cnidaria" rank="species" species="geniculata">
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<emphasis id="B937EA9EFFBDFFCBFEDDFAAEFE21FAD0" bold="true" box="[336,477,1382,1408]" italics="true" pageId="1" pageNumber="86">O. geniculata</emphasis>
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</taxonomicName>
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were gently scraped off with a stiff plastic sheet.
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<collectorName id="26B6535AFFBDFFCBFE0CFA42FE2AFA0D" box="[385,470,1418,1442]" pageId="1" pageNumber="86">Healthy</collectorName>
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tissues located at least
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<quantity id="4CBB9B69FFBDFFCBFD55FA42FCE6FA0E" box="[728,794,1418,1442]" metricMagnitude="-1" metricUnit="m" metricValue="3.0" pageId="1" pageNumber="86" unit="cm" value="30.0">30 cm</quantity>
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from the colony were used as a control.
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<collectorName id="26B6535AFFBDFFCBFDFFFA65FD5AFA6A" box="[626,678,1453,1477]" pageId="1" pageNumber="86">Both</collectorName>
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colonized tissues (blade tissues remaining beneath the colony after the removal of hydrozoans) and healthy tissues collected from many thalli were immediately freeze-dried (
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||
<collectionCode id="ED52AE49FFBDFFCBFD36F9DDFCEAF982" box="[699,790,1557,1581]" pageId="1" pageNumber="86">SFD-SM</collectionCode>
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,
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<collectingCounty id="629D4E00FFBDFFCBFF2BF9FFFDE0F9E0" box="[166,540,1591,1615]" pageId="1" pageNumber="86">Samwon Freezing Engineering Co.</collectingCounty>
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,
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<collectingRegion id="4987F86EFFBDFFCBFDA8F9F0FD91F9E0" box="[549,621,1592,1615]" country="South Korea" name="Busan" pageId="1" pageNumber="86">Busan</collectingRegion>
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||
, Korea), ground to a fine powder, and kept at -70°C before analysis.
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<location id="8E9C6057FFBDFFCBFD68F992FCE6F9DE" LSID="urn:lsid:plazi:treatment:03EA879AFFBDFFCFFF2BFB85FE2CFD81:8E9C6057FFBDFFCBFD68F992FCE6F9DE" box="[741,794,1626,1649]" country="United States of America" county="Samwon Freezing Engineering Co." municipality="St. Louis" name="Most" pageId="1" pageNumber="86" stateProvince="Busan">Most</location>
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||
reagents used in this study were of analytical grade from
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||
<location id="8E9C6057FFBDFFCBFF2BF957FE92F918" LSID="urn:lsid:plazi:treatment:03EA879AFFBDFFCFFF2BFB85FE2CFD81:8E9C6057FFBDFFCBFF2BF957FE92F918" box="[166,366,1695,1719]" country="United States of America" county="Samwon Freezing Engineering Co." municipality="St. Louis" name="Sigma-Aldrich Co." pageId="1" pageNumber="86" stateProvince="Busan">Sigma-Aldrich Co.</location>
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||
,
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||
<collectorName id="26B6535AFFBDFFCBFEF5F957FDF7F918" box="[376,523,1695,1719]" pageId="1" pageNumber="86">
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||
<collectingMunicipality id="6B98ACF6FFBDFFCBFEF5F957FE25F918" box="[376,473,1695,1719]" pageId="1" pageNumber="86">St. Louis</collectingMunicipality>
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||
, MO
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||
</collectorName>
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||
,
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||
<collectingCountry id="F354761CFFBDFFCBFD98F957FDBBF918" box="[533,583,1695,1719]" name="United States of America" pageId="1" pageNumber="86">USA</collectingCountry>
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||
.
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||
</paragraph>
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||
</subSubSection>
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||
<subSubSection id="C3596507FFBDFFCFFF2BF939FE2CFD81" lastPageId="5" lastPageNumber="90" pageId="1" pageNumber="86" type="description">
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<paragraph id="8BFC368CFFBDFFCBFF2BF939FE17F8A0" blockId="1.[166,491,1777,1807]" box="[166,491,1777,1807]" pageId="1" pageNumber="86">
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<emphasis id="B937EA9EFFBDFFCBFF2BF939FE17F8A0" bold="true" box="[166,491,1777,1807]" pageId="1" pageNumber="86">Protein electrophoresis</emphasis>
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||
</paragraph>
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||
<paragraph id="8BFC368CFFBDFFCBFF2BF8F3FB8CFC8E" blockId="1.[166,794,1851,1910]" lastBlockId="1.[824,1452,187,801]" pageId="1" pageNumber="86">
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Protein preparation followed the previous methods of
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<bibRefCitation id="EFD24B7DFFBDFFCBFF4BF896FE36F8D9" author="Getachew, P. & M. A. Hannan & B. H. Nam & J. Y. Cho & Y. K. Hong" box="[198,458,1886,1910]" pageId="1" pageNumber="86" pagination="657 - 664" refId="ref5089" refString="Getachew, P., M. A. Hannan, B. H. Nam, J. Y. Cho and Y. K. Hong. 2014. Induced changes in the proteomic profile of the phaeophyte Saccharina japonica upon colonization by the bryozoan Membranipora membranacea. J. Appl. Phycol. 26: 657 - 664." type="journal article" year="2014">Getachew et al. (2014)</bibRefCitation>
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||
. Briefly, the seaweed powder (
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||
<quantity id="4CBB9B69FFBDFFCBFCB2FF73FC82FF7C" box="[831,894,187,211]" metricMagnitude="-4" metricUnit="kg" metricValue="5.0" pageId="1" pageNumber="86" unit="g" value="0.5">0.5 g</quantity>
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) was homogenized in ten volumes of a lysis solution. Proteins were extracted for 1 h, and used for two-dimensional gel electrophoresis (2-DE). For 2-DE, immobilized pH gradient dry strips (4–10 NL IPG,
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<quantity id="4CBB9B69FFBDFFCBFAEEFEEBFA5BFE94" box="[1379,1447,291,315]" metricMagnitude="-1" metricUnit="m" metricValue="2.4" pageId="1" pageNumber="86" unit="cm" value="24.0">24 cm</quantity>
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||
; Genomine, Pohang,
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||
<collectingCountry id="F354761CFFBDFFCBFBA9FE8EFB9AFEF1" box="[1060,1126,326,350]" name="South Korea" pageId="1" pageNumber="86">Korea</collectingCountry>
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||
) were equilibrated for 14 h, and loaded with 200 µg samples. Isoelectric focusing was performed at 20°C using a Multiphor II electrophoresis unit (GE Healthcare, Little Chalfont,
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||
<collectingCountry id="F354761CFFBDFFCBFACAFE66FA92FE69" box="[1351,1390,430,454]" name="United Kingdom" pageId="1" pageNumber="86">UK</collectingCountry>
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||
). The voltage was increased linearly from 150 to 3500 V over 3 h for sample entry, and the focusing was considered to be complete after 96 kVh. Prior to the second dimension, strips were incubated twice for 10 min each in an equilibration buffer. Equilibrated strips were then inserted onto sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) gels (20 ×
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||
<quantity id="4CBB9B69FFBDFFCBFB3AFD69FAFDFD17" box="[1207,1281,673,697]" metricMagnitude="-1" metricUnit="m" metricValue="2.4" pageId="1" pageNumber="86" unit="cm" value="24.0">24 cm</quantity>
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||
, 10–16%). The SDS-PAGE was run at 20°C for 1700 Vh and silver-stained without fixing, followed by sensitization with glutaraldehyde (
|
||
<bibRefCitation id="EFD24B7DFFBDFFCBFC1AFCC0FB98FC8E" author="Oakley, B. R. & D. R. Kirsch & N. R. Morris" box="[919,1124,776,801]" pageId="1" pageNumber="86" pagination="361 - 363" refId="ref5753" refString="Oakley, B. R., D. R. Kirsch and N. R. Morris. 1980. A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal. Biochem. 105: 361 - 363." type="journal article" year="1980">Oakley et al. 1980</bibRefCitation>
|
||
).
|
||
</paragraph>
|
||
<paragraph id="8BFC368CFFBDFFCBFCB5FCB5FB9CFC34" blockId="1.[824,1120,893,924]" box="[824,1120,893,924]" pageId="1" pageNumber="86">
|
||
<emphasis id="B937EA9EFFBDFFCBFCB5FCB5FB9CFC34" bold="true" box="[824,1120,893,924]" pageId="1" pageNumber="86">Quantitative analysis</emphasis>
|
||
</paragraph>
|
||
<paragraph id="8BFC368CFFBDFFCBFCB5FC00FB4DFB7D" blockId="1.[824,1452,967,1234]" pageId="1" pageNumber="86">
|
||
To evaluate the change in intensity of each protein spot on the 2-D gels, quantitative analysis of digitized images was carried out using PDQuest software (version 7.0; Bio-Rad, Hercules, CA,
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||
<collectingCountry id="F354761CFFBDFFCBFC55FBE7FBF7FBE8" box="[984,1035,1071,1095]" name="United States of America" pageId="1" pageNumber="86">USA</collectingCountry>
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||
). The quantity of each spot was normalized by total valid spot intensity. Protein spots were selected for significant differences in expression of over two-fold or less than half of spot intensity ratio compared with the control or healthy tissues.
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</paragraph>
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||
<paragraph id="8BFC368CFFBDFFCBFCB5FAE6FAD9FAE3" blockId="1.[824,1317,1326,1356]" box="[824,1317,1326,1356]" pageId="1" pageNumber="86">
|
||
<emphasis id="B937EA9EFFBDFFCBFCB5FAE6FAD9FAE3" bold="true" box="[824,1317,1326,1356]" pageId="1" pageNumber="86">Protein digestion and identification</emphasis>
|
||
</paragraph>
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||
<paragraph id="8BFC368CFFBDFFCBFCB5FAB1FBB4F8D9" blockId="1.[824,1452,1401,1910]" pageId="1" pageNumber="86">
|
||
Protein spots were enzymatically digested in gel by the method of
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||
<bibRefCitation id="EFD24B7DFFBDFFCBFC22FA53FB46FA1C" author="Shevchenko, A. & M. Wilm & O. Vorm & M. Mann" box="[943,1210,1435,1459]" pageId="1" pageNumber="86" pagination="850 - 858" refId="ref5898" refString="Shevchenko, A., M. Wilm, O. Vorm and M. Mann. 1996. Mass spectrometric sequencing of proteins from silver-stained polyacrylamide gels. Anal. Chem. 68: 850 - 858." type="journal article" year="1996">Shevchenko et al. (1996)</bibRefCitation>
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||
using porcine trypsin (Promega, Madison, WI,
|
||
<collectingCountry id="F354761CFFBDFFCBFBD9FA76FB7BFA79" box="[1108,1159,1470,1494]" name="United States of America" pageId="1" pageNumber="86">USA</collectingCountry>
|
||
). Gel pieces were washed with 50% acetonitrile, vacuum-dried, and incubated with trypsin (
|
||
<quantity id="4CBB9B69FFBDFFCBFC41F9CBFC00F9B3" box="[972,1020,1539,1564]" metricMagnitude="-12" metricUnit="kg" metricValue="9.0" pageId="1" pageNumber="86" unit="ng" value="9.0">9 ng</quantity>
|
||
<superScript id="7C369BC4FFBDFFCBFB8FF9CCFBDDF9BF" attach="left" box="[1026,1057,1538,1564]" fontSize="6" pageId="1" pageNumber="86">µl-1</superScript>
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||
) in 50 mM ammonium bicarbonate, pH 8.7 for 9 h at 37°C. For the identification of proteins, samples were analyzed using a 4700 Proteomics Analyzer with matrix-assisted laser desorption ionization-time of flight (MALDI-TOF)/TOF™ ion optics (Applied Biosystems, Foster City, CA,
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||
<collectingCountry id="F354761CFFBDFFCBFBA2F979FB9EF966" box="[1071,1122,1713,1737]" name="United States of America" pageId="1" pageNumber="86">USA</collectingCountry>
|
||
). Sequence tag searches were performed via a National Center for Biotechnology Information (NCBI) search using the program Mascot (Matrix Science Ltd., London,
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||
<collectingCountry id="F354761CFFBDFFCBFBCDF8D1FB9FF89E" box="[1088,1123,1817,1841]" name="United Kingdom" pageId="1" pageNumber="86">UK</collectingCountry>
|
||
) and a European Molecular Biology Laboratory (EMBL) search using MS BLAST (
|
||
<bibRefCitation id="EFD24B7DFFBDFFCBFCB2F896FBC1F8D9" author="Shevchenko, A. & S. Sunyaev & A. Loboda & A. Shevchenko & P. Bork & W. Ens & K. G. Standing" box="[831,1085,1886,1910]" pageId="1" pageNumber="86" pagination="1917 - 1926" refId="ref5937" refString="Shevchenko, A., S. Sunyaev, A. Loboda, A. Shevchenko, P. Bork, W. Ens and K. G. Standing. 2001. Charting the proteomes of organisms with unsequenced genomes by MALDI-quadrupole time-of-flight mass spectrometry and BLAST homology searching. Anal. Chem. 73: 1917 - 1926." type="journal article" year="2001">Shevchenko et al. 2001</bibRefCitation>
|
||
).
|
||
</paragraph>
|
||
<paragraph id="8BFC368CFFBEFFC8FF05FF72FEF6FF4F" blockId="2.[136,266,186,224]" box="[136,266,186,224]" pageId="2" pageNumber="87">
|
||
<heading id="D0B481E0FFBEFFC8FF05FF72FEF6FF4F" box="[136,266,186,224]" fontSize="15" level="1" pageId="2" pageNumber="87" reason="1">
|
||
<emphasis id="B937EA9EFFBEFFC8FF05FF72FEF6FF4F" bold="true" box="[136,266,186,224]" pageId="2" pageNumber="87">Results</emphasis>
|
||
</heading>
|
||
</paragraph>
|
||
<paragraph id="8BFC368CFFBEFFC8FF05FEC9FA83FD16" blockId="2.[136,764,255,731]" lastBlockId="2.[794,1422,187,1944]" pageId="2" pageNumber="87">
|
||
The stoloniferous hydrozoan
|
||
<taxonomicName id="4C434D0FFFBEFFC8FE44FF37FD7AFEB7" box="[457,646,255,281]" class="Hydrozoa" family="Campanulariidae" genus="Obelia" kingdom="Animalia" order="Leptothecata" pageId="2" pageNumber="87" phylum="Cnidaria" rank="species" species="geniculata">
|
||
<emphasis id="B937EA9EFFBEFFC8FE44FF37FD7AFEB7" bold="true" box="[457,646,255,281]" italics="true" pageId="2" pageNumber="87">Obelia geniculata</emphasis>
|
||
</taxonomicName>
|
||
was widespread on blades of late-harvested
|
||
<taxonomicName id="4C434D0FFFBEFFC8FD9BFEEAFD08FE94" baseAuthorityName="Park and Hwang" baseAuthorityYear="2012" box="[534,756,290,316]" class="Phaeophyceae" family="Laminariaceae" genus="Saccharina" kingdom="Chromista" order="Laminariales" pageId="2" pageNumber="87" phylum="Ochrophyta" rank="species" species="japonica">
|
||
<emphasis id="B937EA9EFFBEFFC8FD9BFEEAFD08FE94" bold="true" box="[534,756,290,316]" italics="true" pageId="2" pageNumber="87">Saccharina japonica</emphasis>
|
||
</taxonomicName>
|
||
. Blade parts with hydrozoan-colonies and healthy tissues were used to isolate proteins induced by
|
||
<taxonomicName id="4C434D0FFFBEFFC8FDE4FEAFFD07FE2F" box="[617,763,359,385]" class="Hydrozoa" family="Campanulariidae" genus="Obelia" kingdom="Animalia" order="Leptothecata" pageId="2" pageNumber="87" phylum="Cnidaria" rank="species" species="geniculata">
|
||
<emphasis id="B937EA9EFFBEFFC8FDE4FEAFFD07FE2F" bold="true" box="[617,763,359,385]" italics="true" pageId="2" pageNumber="87">O. geniculata</emphasis>
|
||
</taxonomicName>
|
||
infection. The protein isolation from tissues was replicated and optimized to confirm the differently expressed protein profiles. In the hydrozoan-colonized tissues, 107 protein spots were detected on a 2-DE gel plate, while 75 spots were detected in the healthy tissues (
|
||
<figureCitation id="13782A09FFBEFFC8FD1FFDDEFD11FD81" box="[658,749,534,558]" captionStart="Figure 1" captionStartId="2.[136,193,1835,1855]" captionTargetBox="[136,763,815,1804]" captionTargetId="figure-140@2.[136,763,814,1804]" captionTargetPageId="2" captionText="Figure 1: Two-dimensional gel electrophoresis profiles of late-harvested Saccharina japonica. (A) Healthy tissues. (B) Hydrozoan-colonized tissues. The separated proteins were visualized by silver staining." figureDoi="http://doi.org/10.5281/zenodo.11360031" httpUri="https://zenodo.org/record/11360031/files/figure.png" pageId="2" pageNumber="87">Figure 1</figureCitation>
|
||
). Out of the 107 spots in colonized tissues, 105 had different expression levels between the healthy and colonized tissues; 77 and 28 spots were up- and down-regulated, respectively, upon
|
||
<taxonomicName id="4C434D0FFFBEFFC8FED7FD57FE15FD17" box="[346,489,671,697]" class="Hydrozoa" family="Campanulariidae" genus="Obelia" kingdom="Animalia" order="Leptothecata" pageId="2" pageNumber="87" phylum="Cnidaria" rank="species" species="geniculata">
|
||
<emphasis id="B937EA9EFFBEFFC8FED7FD57FE15FD17" bold="true" box="[346,489,671,697]" italics="true" pageId="2" pageNumber="87">O. geniculata</emphasis>
|
||
</taxonomicName>
|
||
colonization. Out of the 77 up-regulated spots, 30 had more than twice the spot intensity compared with the healthy tissues. Out of the 28 down-regulated spots, 22 had less than half the spot intensity compared with the healthy tissues. Among these 52 (30+22) spots, 38 clear and abundant spots, which appeared constantly in replicated experiments, were selected and subjected to protein analysis. Through a database search of proteins from algae, land plants, and bacteria, 21 spots were identified, of which two were mixtures of two proteins. The identities of these 23 proteins and their molecular weight (MW), isoelectric point (p
|
||
<emphasis id="B937EA9EFFBEFFC8FAF1FE3AFA78FDA4" bold="true" box="[1404,1412,498,523]" italics="true" pageId="2" pageNumber="87">I</emphasis>
|
||
) values, and functions are summarized in
|
||
<tableCitation id="C6C10337FFBEFFC8FB6EFDDEFACCFD81" box="[1251,1328,534,558]" captionStart="Table 1" captionStartId="3.[136,186,435,458]" captionTargetBox="[136,1900,526,1680]" captionText="Table 1: Identified proteins in the hydrozoan-colonized tissues and healthy tissues of the late-harvested Saccharina japonica. Ratio of spot intensity was expressed by colonized tissues per healthy tissues." pageId="2" pageNumber="87">Table 1</tableCitation>
|
||
. Among them, 7 and 16 identified proteins were significantly up-and down-regulated, respectively. By searching the NCBI and EMBL databases, 17 and 18 proteins, respectively, were confidently identified. Twelve proteins overlapped.
|
||
</paragraph>
|
||
<paragraph id="8BFC368CFFBEFFCFFCCAFD0BFE2CFD81" blockId="2.[794,1422,187,1944]" lastBlockId="5.[166,794,187,558]" lastPageId="5" lastPageNumber="90" pageId="2" pageNumber="87">
|
||
The identified 23 proteins were cell-division cycle 46/ minichromosome maintenance protein 5 (Cdc46/Mcm5), glutamyl-tRNA reductase (GluTR), microcompartments protein (MCP), carboxysome shell peptide (CsoS), biotin synthesis protein (bioC), serine/arginine-rich splicing factor (SRSF), two-component response regulator (PilR), chloroplast phosphoglycerate kinase (PGK), expansin 6 (EXPA6), translation initiation factor 3 (IF3), calcium/ calmodulin-dependent protein kinase II inhibitor 2 (CaMK2N2), 50S ribosomal protein L1P (rpl1P), transmembrane protein (TP), protoporphyrinogen oxidase (PPOX), dual oxidase 2 like (DUOX2), PIH1 domain-containing protein 2 (Hsp90), GTPase-activating protein alpha (GAPα), threonyl-tRNA synthetase (TARS), flavanone 3-hydroxylase (F3H), uncoupling protein 3 (UCP3), vanadium-dependent bromoperoxidase 7 (vBPO7), peptide chain release factor 1 (Prf1), and interaptin in a database search of both NCBI and EMBL. Among the identified proteins, two up-regulated proteins (Cdc46/Mcm5 and GluTR) were mostly expressed only in the hydrozoan-colonized tissues but were rare in the healthy tissues (
|
||
<figureCitation id="13782A09FFBEFFC8FB7AFAB0FAAEFA3F" box="[1271,1362,1400,1424]" captionStart="Figure 2" captionStartId="4.[794,851,388,408]" captionTargetBox="[794,1421,174,357]" captionTargetId="figure-191@4.[794,1421,174,357]" captionTargetPageId="4" captionText="Figure 2: A close-up view of 2-dimensional electrophoresis gels showing the identified up-regulated proteins (indicated by arrows) found mostly in hydrozoan-colonized tissues but rare in healthy tissues. (A) Healthy tissues. (B) Hydrozoan-colonized tissues." figureDoi="http://doi.org/10.5281/zenodo.11360033" httpUri="https://zenodo.org/record/11360033/files/figure.png" pageId="2" pageNumber="87">Figure 2</figureCitation>
|
||
). The Cdc46/Mcm5, related to stress control, showed sharply increased spot intensity or protein amount; approximately 1308-fold more in colonized tissues than in healthy tissues. The spot intensity of photosynthesis-related GluTR also increased sharply by approximately 277-fold. Five proteins (MCP, CsoS, bioC, SRSF and PilR) in photosynthesis, stress control, and signal transduction were significantly up-regulated by approximately 3–11-fold in hydrozoan-colonized tissues (
|
||
<figureCitation id="13782A09FFBEFFC8FBEBF978FB39F967" box="[1126,1221,1712,1736]" captionStart="Figure 3" captionStartId="4.[794,851,1044,1064]" captionTargetBox="[794,1420,634,1012]" captionTargetId="figure-231@4.[794,1420,634,1013]" captionTargetPageId="4" captionText="Figure 3: A close-up view of 2-dimensional electrophoresis gels showing the identified up-regulated proteins (indicated by arrows) altered by hydrozoan colonization. (A) Healthy tissues. (B) Hydrozoan-colonized tissues." figureDoi="http://doi.org/10.5281/zenodo.11360035" httpUri="https://zenodo.org/record/11360035/files/figure.png" pageId="2" pageNumber="87">Figure 3</figureCitation>
|
||
). Meanwhile, five down-regulated proteins (PGK, EXPA6, IF3, CAMK2N2 and rpl1P), which were found mostly in healthy tissues but were rare in hydrozoan-colonized tissues, were related to photosynthesis, cell growth, stress control and signal transduction in a database search (
|
||
<figureCitation id="13782A09FFBEFFC8FB13F895FB07F8D9" box="[1182,1275,1885,1910]" captionStart="Figure 4" captionStartId="4.[794,851,1806,1826]" captionTargetBox="[794,1415,1260,1775]" captionTargetId="figure-271@4.[794,1415,1259,1775]" captionTargetPageId="4" captionText="Figure 4: A close-up view of 2-dimensional electrophoresis gels showing the identified down-regulated proteins (indicated by arrows) found mostly in healthy tissues but rare in hydrozoancolonized tissues. (A) Healthy tissues. (B) Hydrozoan-colonized tissues." figureDoi="http://doi.org/10.5281/zenodo.11360037" httpUri="https://zenodo.org/record/11360037/files/figure.png" pageId="2" pageNumber="87">Figure 4</figureCitation>
|
||
). Eleven proteins (TP, PPOX, DUOX2, Hsp90, GAPα, TARS, F3H, UCP3,
|
||
<emphasis id="B937EA9EFFB8FFCEFF04F915FF5CF94D" bold="true" box="[137,160,1757,1762]" pageId="4" pageNumber="89">)</emphasis>
|
||
vBPO7, Prf1, and interaptin), related to signal transduction, defense response, protein metabolism, stress control, photosynthesis and the cytoskeleton were significantly down-regulated by 0.1–0.5-fold in the hydrozoan-colonized tissues (
|
||
<figureCitation id="13782A09FFB9FFCFFECAFE8EFE5AFEF1" box="[327,422,326,350]" captionStart="Figure 5" captionStartId="5.[166,223,1835,1855]" captionTargetBox="[166,787,991,1804]" captionTargetId="figure-140@5.[166,788,990,1804]" captionTargetPageId="5" captionText="Figure 5: A close-up view of 2-dimensional electrophoresis gels showing the identified down-regulated proteins (indicated by arrows) altered by hydrozoan colonization. (A) Healthy tissues. (B) Hydrozoan-colonized tissues." figureDoi="http://doi.org/10.5281/zenodo.11360039" httpUri="https://zenodo.org/record/11360039/files/figure.png" pageId="5" pageNumber="90">Figure 5</figureCitation>
|
||
). From the 23 proteins identified through a homology-based cross-species database, we found that six proteins were related to stress control, five proteins to signal transduction, five proteins to photosynthesis, two proteins to protein metabolism, two proteins to defense response, two proteins to cell growth, and one protein to the cytoskeleton.
|
||
</paragraph>
|
||
</subSubSection>
|
||
</treatment>
|
||
</document> |