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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en"><italic>Citrus Tristeza Virus</italic>
: Survival at the Edge of the Movement Continuum<xref ref-type="fn" rid="fn3">▿</xref>
</title>
<author><name sortKey="Folimonova, Svetlana Y" sort="Folimonova, Svetlana Y" uniqKey="Folimonova S" first="Svetlana Y." last="Folimonova">Svetlana Y. Folimonova</name>
<affiliation><nlm:aff id="aff0">Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, Florida 33850</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Folimonov, Alexey S" sort="Folimonov, Alexey S" uniqKey="Folimonov A" first="Alexey S." last="Folimonov">Alexey S. Folimonov</name>
<affiliation><nlm:aff id="aff0">Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, Florida 33850</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Tatineni, Satyanarayana" sort="Tatineni, Satyanarayana" uniqKey="Tatineni S" first="Satyanarayana" last="Tatineni">Satyanarayana Tatineni</name>
<affiliation><nlm:aff id="aff0">Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, Florida 33850</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Dawson, William O" sort="Dawson, William O" uniqKey="Dawson W" first="William O." last="Dawson">William O. Dawson</name>
<affiliation><nlm:aff id="aff0">Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, Florida 33850</nlm:aff>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PMC</idno>
<idno type="pmid">18434397</idno>
<idno type="pmc">2447058</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2447058</idno>
<idno type="RBID">PMC:2447058</idno>
<idno type="doi">10.1128/JVI.00515-08</idno>
<date when="2008">2008</date>
<idno type="wicri:Area/Pmc/Corpus">000777</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main"><italic>Citrus Tristeza Virus</italic>
: Survival at the Edge of the Movement Continuum<xref ref-type="fn" rid="fn3">▿</xref>
</title>
<author><name sortKey="Folimonova, Svetlana Y" sort="Folimonova, Svetlana Y" uniqKey="Folimonova S" first="Svetlana Y." last="Folimonova">Svetlana Y. Folimonova</name>
<affiliation><nlm:aff id="aff0">Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, Florida 33850</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Folimonov, Alexey S" sort="Folimonov, Alexey S" uniqKey="Folimonov A" first="Alexey S." last="Folimonov">Alexey S. Folimonov</name>
<affiliation><nlm:aff id="aff0">Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, Florida 33850</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Tatineni, Satyanarayana" sort="Tatineni, Satyanarayana" uniqKey="Tatineni S" first="Satyanarayana" last="Tatineni">Satyanarayana Tatineni</name>
<affiliation><nlm:aff id="aff0">Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, Florida 33850</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Dawson, William O" sort="Dawson, William O" uniqKey="Dawson W" first="William O." last="Dawson">William O. Dawson</name>
<affiliation><nlm:aff id="aff0">Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, Florida 33850</nlm:aff>
</affiliation>
</author>
</analytic>
<series><title level="j">Journal of Virology</title>
<idno type="ISSN">0022-538X</idno>
<idno type="eISSN">1098-5514</idno>
<imprint><date when="2008">2008</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><p>Systemic invasion of plants by viruses is thought to involve two processes: cell-to-cell movement between adjacent cells and long-distance movement that allows the virus to rapidly move through sieve elements and unload at the growing parts of the plant. There is a continuum of proportions of these processes that determines the degrees of systemic infection of different plants by different viruses. We examined the systemic distribution of <italic>Citrus tristeza virus</italic>
(CTV) in citrus species with a range of susceptibilities. By using a “pure” culture of CTV from a cDNA clone and green fluorescent protein-labeled virus we show that both cell-to-cell and long-distance movement are unusually limited, and the degree of limitation varies depending on the citrus host. In the more-susceptible hosts CTV infected only a small portion of phloem-associated cells, and moreover, the number of infection sites in less-susceptible citrus species was substantially decreased further, indicating that long-distance movement was reduced in those hosts. Analysis of infection foci in the two most differential citrus species, <italic>Citrus macrophylla</italic>
and sour orange, revealed that in the more-susceptible host the infection foci were composed of a cluster of multiple cells, while in the less-susceptible host infection foci were usually single cells, suggesting that essentially no cell-to-cell movement occurred in the latter host. Thus, CTV in sour orange represents a pattern of systemic infection in which the virus appears to function with only the long-distance movement mechanism, yet is able to survive in nature.</p>
</div>
</front>
</TEI>
<pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Virol</journal-id>
<journal-id journal-id-type="publisher-id">jvi</journal-id>
<journal-title>Journal of Virology</journal-title>
<issn pub-type="ppub">0022-538X</issn>
<issn pub-type="epub">1098-5514</issn>
<publisher><publisher-name>American Society for Microbiology (ASM)</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">18434397</article-id>
<article-id pub-id-type="pmc">2447058</article-id>
<article-id pub-id-type="publisher-id">0515-08</article-id>
<article-id pub-id-type="doi">10.1128/JVI.00515-08</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Pathogenesis and Immunity</subject>
</subj-group>
</article-categories>
<title-group><article-title><italic>Citrus Tristeza Virus</italic>
: Survival at the Edge of the Movement Continuum<xref ref-type="fn" rid="fn3">▿</xref>
</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Folimonova</surname>
<given-names>Svetlana Y.</given-names>
</name>
<xref ref-type="aff" rid="aff0"></xref>
</contrib>
<contrib contrib-type="author"><name><surname>Folimonov</surname>
<given-names>Alexey S.</given-names>
</name>
<xref ref-type="aff" rid="aff0"></xref>
<xref ref-type="fn" rid="fn1">†</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Tatineni</surname>
<given-names>Satyanarayana</given-names>
</name>
<xref ref-type="aff" rid="aff0"></xref>
<xref ref-type="fn" rid="fn2">‡</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Dawson</surname>
<given-names>William O.</given-names>
</name>
<xref ref-type="aff" rid="aff0"></xref>
<xref ref-type="corresp" rid="cor1">*</xref>
</contrib>
</contrib-group>
<aff id="aff0">Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, Florida 33850</aff>
<author-notes><fn id="cor1"><label>*</label>
<p>Corresponding author. Mailing address: Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, FL 33850. Phone: (863) 956-1151. Fax: (863) 956-4631. E-mail: <email>wodtmv@crec.ifas.ufl.edu</email>
</p>
</fn>
<fn id="fn1"><label>†</label>
<p>Present address: Center Bioengineering, Russian Academy of Sciences, Moscow 117312, Russia.</p>
</fn>
<fn id="fn2"><label>‡</label>
<p>Present address: U.S. Department of Agriculture Agricultural Research Service and Department of Plant Pathology, University of Nebraska, Lincoln, NE 68583.</p>
</fn>
</author-notes>
<pub-date pub-type="ppub"><month>7</month>
<year>2008</year>
</pub-date>
<pub-date pub-type="epub"><day>23</day>
<month>4</month>
<year>2008</year>
</pub-date>
<volume>82</volume>
<issue>13</issue>
<fpage>6546</fpage>
<lpage>6556</lpage>
<history><date date-type="received"><day>7</day>
<month>3</month>
<year>2008</year>
</date>
<date date-type="accepted"><day>14</day>
<month>4</month>
<year>2008</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2008, American Society for Microbiology</copyright-statement>
</permissions>
<self-uri xlink:title="pdf" xlink:href="zjv01308006546.pdf"></self-uri>
<abstract><p>Systemic invasion of plants by viruses is thought to involve two processes: cell-to-cell movement between adjacent cells and long-distance movement that allows the virus to rapidly move through sieve elements and unload at the growing parts of the plant. There is a continuum of proportions of these processes that determines the degrees of systemic infection of different plants by different viruses. We examined the systemic distribution of <italic>Citrus tristeza virus</italic>
(CTV) in citrus species with a range of susceptibilities. By using a “pure” culture of CTV from a cDNA clone and green fluorescent protein-labeled virus we show that both cell-to-cell and long-distance movement are unusually limited, and the degree of limitation varies depending on the citrus host. In the more-susceptible hosts CTV infected only a small portion of phloem-associated cells, and moreover, the number of infection sites in less-susceptible citrus species was substantially decreased further, indicating that long-distance movement was reduced in those hosts. Analysis of infection foci in the two most differential citrus species, <italic>Citrus macrophylla</italic>
and sour orange, revealed that in the more-susceptible host the infection foci were composed of a cluster of multiple cells, while in the less-susceptible host infection foci were usually single cells, suggesting that essentially no cell-to-cell movement occurred in the latter host. Thus, CTV in sour orange represents a pattern of systemic infection in which the virus appears to function with only the long-distance movement mechanism, yet is able to survive in nature.</p>
</abstract>
</article-meta>
</front>
<floats-wrap><fig position="float" id="f1"><label>FIG. 1.</label>
<caption><p>(A) Schematic diagram of the genome organization of wild-type CTV (CTV9R) and its derivative encoding GFP. The open boxes represent ORFs and their translation products. PRO, papain-like protease domain; MT, methyltransferase; HEL, helicase; RdRp, an RNA-dependent RNA polymerase; HSP70h, HSP70 homolog. Bent arrows indicate positions of BYV (<italic>B<sub>CP</sub>
</italic>
) or CTV CP (<italic>C<sub>CP</sub>
</italic>
) sgRNA controller elements. Inserted elements are shown in gray. (B) Replication of CTV9R (lane 1) or CTV-BC5/GFP (lane 2) in <italic>N. benthamiana</italic>
mesophyll protoplasts inoculated with virions extracted from infected <italic>C. macrophylla</italic>
seedlings. Total RNA was isolated from protoplasts 4 days postinoculation. Northern blot hybridizations were carried out using CTV 3′ positive RNA strand-specific riboprobe. Positions of sgRNAs are shown.</p>
</caption>
<graphic xlink:href="zjv0130807580001"></graphic>
</fig>
<fig position="float" id="f2"><label>FIG. 2.</label>
<caption><p>Transmission electron micrographs showing phloem cells in a petiole of <italic>C. macrophylla</italic>
infected with CTV9R. (A) Groups of phloem cells at lower magnification. Infected cells are indicated with letters B, C, and D, corresponding to images in subsequent panels. (B to D) Areas from the cells shown in panel at higher magnification. Viral arrays were labeled with polyclonal CTV-specific antibodies used as primary antibodies and secondary antibodies conjugated with 10-nm gold particles. No labeling was detected in noninfected cells.</p>
</caption>
<graphic xlink:href="zjv0130807580002"></graphic>
</fig>
<fig position="float" id="f3"><label>FIG. 3.</label>
<caption><p>Transmission electron micrographs showing phloem cells in a petiole of Mexican lime infected with CTV9R. (A) Groups of phloem cells at lower magnification. Infected cells are indicated with letters B and C, corresponding to images in those panels. (B and C) Areas from the infected cells shown in panel A at higher magnification. Masses of virions were labeled with polyclonal CTV-specific antibodies used as primary antibodies and secondary antibodies conjugated with 10-nm gold particles.</p>
</caption>
<graphic xlink:href="zjv0130807580003"></graphic>
</fig>
<fig position="float" id="f4"><label>FIG. 4.</label>
<caption><p>Detection of GFP fluorescence in phloem-associated cells on the internal surface of bark of <italic>C. macrophylla</italic>
(CM), Mexican lime (ML), sweet orange Madam Vinous (MV), Duncan grapefruit (DG), and sour orange (SO) trees infected with CTV-BC5/GFP at 6 weeks after inoculation (micrographs in the top row). Indicated areas of fluorescing cells in bark of <italic>C. macrophylla</italic>
and sour orange are shown at higher magnification (bottom row) to demonstrate dramatic differences in virus distribution in these two species.</p>
</caption>
<graphic xlink:href="zjv0130807580004"></graphic>
</fig>
<fig position="float" id="f5"><label>FIG. 5.</label>
<caption><p>(A) Detection of GFP fluorescence in phloem-associated cells on the internal surface of bark of <italic>C. macrophylla</italic>
and sour orange shoots from a “duplex” plant created by grafting of sour orange onto a <italic>C. macrophylla</italic>
tree infected with CTV-BC5/GFP. The image was taken at 6 months after the graft was done. (B) Scheme of the “bark patch” experiment. A small piece of bark was excised from the stem of a <italic>C. macrophylla</italic>
tree and replaced with a bark piece of an equal size from a sour orange tree. The substituted bark patch was then allowed to become grafted in place. Six months after inoculation with GFP-expressing CTV, fluorescence was observed in the bark tissue excised from <italic>C. macrophylla</italic>
at a region containing grafted bark patch of sour orange. CM, <italic>C. macrophylla</italic>
; SO, sour orange.</p>
</caption>
<graphic xlink:href="zjv0130807580005"></graphic>
</fig>
<fig position="float" id="f6"><label>FIG. 6.</label>
<caption><p>Confocal laser scanning microscope images of individual infection foci in the bark of several <italic>C. macrophylla</italic>
(A) or sour orange (B) plants at 6 weeks after inoculation with CTV-BC5/GFP. The four panels represent replicas from different plants.</p>
</caption>
<graphic xlink:href="zjv0130807580006"></graphic>
</fig>
<fig position="float" id="f7"><label>FIG. 7.</label>
<caption><p>Light microscope images of infection foci in the bark of <italic>C. macrophylla</italic>
(A) or sour orange (B) infected with wild-type CTV at 6 weeks after inoculation. Tissue sections of fixed samples from infected trees were labeled with CTV-specific antibodies as primary antibodies and antibodies with attached 10-nm gold particles as secondary antibodies. To visualize infected cells at the light microscopy level, labeling was further enhanced with the silver enhancement technique.</p>
</caption>
<graphic xlink:href="zjv0130807580007"></graphic>
</fig>
<table-wrap position="float" id="t1"><label>TABLE 1.</label>
<caption><p>Analysis of CTV accumulation in five citrus species</p>
</caption>
<table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="2" align="center" valign="middle">Exptl group</th>
<th colspan="5" rowspan="1" align="center" valign="bottom">CTV titer<xref ref-type="table-fn" rid="t1fn1"><italic>a</italic>
</xref>
in:
<hr></hr>
</th>
</tr>
<tr><th colspan="1" rowspan="1" align="center" valign="bottom"><italic>Citrus macrophylla</italic>
</th>
<th colspan="1" rowspan="1" align="center" valign="bottom">Mexican lime</th>
<th colspan="1" rowspan="1" align="center" valign="bottom">Sweet orange Madam Vinous</th>
<th colspan="1" rowspan="1" align="center" valign="bottom">Duncan grapefruit</th>
<th colspan="1" rowspan="1" align="center" valign="bottom">Sour orange</th>
</tr>
</thead>
<tbody><tr><td colspan="1" rowspan="1" align="left" valign="top">CTV9R infected</td>
<td colspan="1" rowspan="1" align="center" valign="top">3.08 ± 0.015</td>
<td colspan="1" rowspan="1" align="center" valign="top">2.95 ± 0.021</td>
<td colspan="1" rowspan="1" align="center" valign="top">0.92 ± 0.065</td>
<td colspan="1" rowspan="1" align="center" valign="top">0.54 ± 0.030</td>
<td colspan="1" rowspan="1" align="center" valign="top">0.32 ± 0.015</td>
</tr>
<tr><td colspan="1" rowspan="1" align="left" valign="top">Healthy</td>
<td colspan="1" rowspan="1" align="center" valign="top">0.10 ± 0.004</td>
<td colspan="1" rowspan="1" align="center" valign="top">0.09 ± 0.003</td>
<td colspan="1" rowspan="1" align="center" valign="top">0.08 ± 0.008</td>
<td colspan="1" rowspan="1" align="center" valign="top">0.09 ± 0.009</td>
<td colspan="1" rowspan="1" align="center" valign="top">0.07 ± 0.008</td>
</tr>
</tbody>
</table>
<table-wrap-foot><fn id="t1fn1"><label>a</label>
<p>CTV9R-infected trees were assayed at 6 weeks postinoculation by double antibody sandwich indirect ELISA using CTV-specific 908 IgG as trapping antibody at a 1-μg/ml concentration and ECTV 172 monoclonal antibody as detecting antibody at a 1:50,000 dilution. ELISA values (<italic>A</italic>
<sub>405</sub>
) are averages for three plants ± standard deviations.</p>
</fn>
</table-wrap-foot>
</table-wrap>
</floats-wrap>
</pmc>
</record>
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