Spiralin Is Not Essential for Helicity, Motility, or Pathogenicity but Is Required for Efficient Transmission of Spiroplasma citri by Its Leafhopper Vector Circulifer haematoceps
Identifieur interne : 000223 ( Ncbi/Curation ); précédent : 000222; suivant : 000224Spiralin Is Not Essential for Helicity, Motility, or Pathogenicity but Is Required for Efficient Transmission of Spiroplasma citri by Its Leafhopper Vector Circulifer haematoceps
Auteurs : Sybille Duret [France] ; Nathalie Berho [France] ; Jean-Luc Danet [France] ; Monique Garnier [France] ; Joël Renaudin [France]Source :
- Applied and Environmental Microbiology [ 0099-2240 ] ; 2003.
Abstract
Spiralin is the most abundant protein at the surface of the plant pathogenic mollicute
Url:
DOI: 10.1128/AEM.69.10.6225-6234.2003
PubMed: 14532084
PubMed Central: 201218
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PMC:201218Le document en format XML
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by Its Leafhopper Vector <italic>Circulifer haematoceps</italic>
</title>
<author><name sortKey="Duret, Sybille" sort="Duret, Sybille" uniqKey="Duret S" first="Sybille" last="Duret">Sybille Duret</name>
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<country xml:lang="fr">France</country>
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<author><name sortKey="Garnier, Monique" sort="Garnier, Monique" uniqKey="Garnier M" first="Monique" last="Garnier">Monique Garnier</name>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Spiralin Is Not Essential for Helicity, Motility, or Pathogenicity but Is Required for Efficient Transmission of <italic>Spiroplasma citri</italic>
by Its Leafhopper Vector <italic>Circulifer haematoceps</italic>
</title>
<author><name sortKey="Duret, Sybille" sort="Duret, Sybille" uniqKey="Duret S" first="Sybille" last="Duret">Sybille Duret</name>
<affiliation wicri:level="3"><nlm:aff id="aff0">UMR Génomique Développement et Pouvoir Pathogène, IBVM, Centre INRA de Bordeaux, 33883 Villenave d'Ornon Cedex, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>UMR Génomique Développement et Pouvoir Pathogène, IBVM, Centre INRA de Bordeaux, 33883 Villenave d'Ornon Cedex</wicri:regionArea>
<placeName><region type="region" nuts="2">Nouvelle-Aquitaine</region>
<region type="old region" nuts="2">Aquitaine</region>
<settlement type="city">Villenave d'Ornon</settlement>
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<author><name sortKey="Berho, Nathalie" sort="Berho, Nathalie" uniqKey="Berho N" first="Nathalie" last="Berho">Nathalie Berho</name>
<affiliation wicri:level="3"><nlm:aff id="aff0">UMR Génomique Développement et Pouvoir Pathogène, IBVM, Centre INRA de Bordeaux, 33883 Villenave d'Ornon Cedex, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>UMR Génomique Développement et Pouvoir Pathogène, IBVM, Centre INRA de Bordeaux, 33883 Villenave d'Ornon Cedex</wicri:regionArea>
<placeName><region type="region" nuts="2">Nouvelle-Aquitaine</region>
<region type="old region" nuts="2">Aquitaine</region>
<settlement type="city">Villenave d'Ornon</settlement>
</placeName>
</affiliation>
</author>
<author><name sortKey="Danet, Jean Luc" sort="Danet, Jean Luc" uniqKey="Danet J" first="Jean-Luc" last="Danet">Jean-Luc Danet</name>
<affiliation wicri:level="3"><nlm:aff id="aff0">UMR Génomique Développement et Pouvoir Pathogène, IBVM, Centre INRA de Bordeaux, 33883 Villenave d'Ornon Cedex, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>UMR Génomique Développement et Pouvoir Pathogène, IBVM, Centre INRA de Bordeaux, 33883 Villenave d'Ornon Cedex</wicri:regionArea>
<placeName><region type="region" nuts="2">Nouvelle-Aquitaine</region>
<region type="old region" nuts="2">Aquitaine</region>
<settlement type="city">Villenave d'Ornon</settlement>
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<author><name sortKey="Garnier, Monique" sort="Garnier, Monique" uniqKey="Garnier M" first="Monique" last="Garnier">Monique Garnier</name>
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<settlement type="city">Villenave d'Ornon</settlement>
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<author><name sortKey="Renaudin, Joel" sort="Renaudin, Joel" uniqKey="Renaudin J" first="Joël" last="Renaudin">Joël Renaudin</name>
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<country xml:lang="fr">France</country>
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<series><title level="j">Applied and Environmental Microbiology</title>
<idno type="ISSN">0099-2240</idno>
<idno type="eISSN">1098-5336</idno>
<imprint><date when="2003">2003</date>
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<front><div type="abstract" xml:lang="en"><p>Spiralin is the most abundant protein at the surface of the plant pathogenic mollicute <italic>Spiroplasma citri</italic>
and hence might play a role in the interactions of the spiroplasma with its host plant and/or its insect vector. To study spiralin function, mutants were produced by inactivating the spiralin gene through homologous recombination. A spiralin-green fluorescent protein (GFP) translational fusion was engineered and introduced into <italic>S. citri</italic>
by using an <italic>oriC</italic>
-based targeting vector. According to the strategy used, integration of the plasmid by a single-crossover recombination at the spiralin gene resulted in the expression of the spiralin-GFP fusion protein. Two distinct mutants were isolated. Western and colony immunoblot analyses showed that one mutant (GII3-9a5) did produce the spiralin-GFP fusion protein, which was found not to fluoresce, whereas the other (GII3-9a2) produced neither the fusion protein nor the wild-type spiralin. Both mutants displayed helical morphology and motility, similarly to the wild-type strain GII-3. Genomic DNA analyses revealed that GII3-9a5 was unstable and that GII3-9a2 was probably derived from GII3-9a5 by a double-crossover recombination between plasmid sequences integrated into the GII3-9a5 chromosome and free plasmid. When injected into the leafhopper vector <italic>Circulifer haematoceps</italic>
, the spiralinless mutant GII3-9a2 multiplied to high titers in the insects (1.1 × 10<sup>6</sup>
to 2.8 × 10<sup>6</sup>
CFU/insect) but was transmitted to the host plant 100 times less efficiently than the wild-type strain. As a result, not all plants were infected, and symptom production in these plants was delayed for 2 to 4 weeks compared to that in the wild-type strain. In the infected plants however, the mutant multiplied to high titers (1.2 × 10<sup>6</sup>
to 1.4 × 10<sup>7</sup>
CFU/g of midribs) and produced the typical symptoms of the disease. These results indicate that spiralin is not essential for pathogenicity but is required for efficient transmission of <italic>S. citri</italic>
by its insect vector.</p>
</div>
</front>
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