Journal article Open Access

Towards a sustainable strategy for Xylella fastidiosa control

G. D'Attoma, M. Morelli, S. Cicco, M. Saponari, P. Saldarelli

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    <subfield code="a">xylella fastidiosa, DSF, quorum sensing, olive</subfield>
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    <subfield code="a">G. D'Attoma, M. Morelli, S. Cicco, M. Saponari, P. Saldarelli</subfield>
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    <subfield code="a">Towards a sustainable strategy for Xylella fastidiosa control</subfield>
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    <subfield code="a">Pest Organisms Threatening Europe</subfield>
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    <subfield code="a">&lt;p&gt;&lt;em&gt;Xylella fastidiosa&lt;/em&gt; (&lt;em&gt;Xf&lt;/em&gt;) is a xylem-limited bacterium, regulated as a quarantine pest, that is causing a devastating disease on olive crops in the southern area of Apulia (Italy) and whose potential spread in the Mediterranean area poses a severe threat to EU agriculture and landscape environment.&lt;/p&gt;

&lt;p&gt;&lt;em&gt;Xf&lt;/em&gt; virulence is related to the expression of a cluster of &lt;em&gt;rpf&lt;/em&gt; (regulation of pathogenicity factors) genes responsible for a signalling system based on small fatty acid molecules called DSF (Diffusible Signalling Factor). Since DSF regulation is involved in pathogenicity traits of &lt;em&gt;Xf&lt;/em&gt; and biofilm formation, a “pathogen&lt;strong&gt;-&lt;/strong&gt;confusion” strategy, based on the alteration of DSF levels &lt;em&gt;in planta&lt;/em&gt;,&lt;em&gt; &lt;/em&gt;has been proposed to contrast bacterial infection. In grapevine, the strategy is based on the transgenic expression of the &lt;em&gt;rpfF&lt;/em&gt; gene, which encodes the DSF-synthase.&lt;/p&gt;

&lt;p&gt;We are attempting to express the &lt;em&gt;rpfF&lt;/em&gt; gene of the&lt;em&gt; &lt;/em&gt;olive-infecting &lt;em&gt;Xf&lt;/em&gt; strain CoDiRO in the heterologous &lt;em&gt;Escherichia coli &lt;/em&gt;system. The gene product has been successfully detected by Western blot analysis in cell protein extracts. Chemical characterisation by Gas Chromatography-Mass Spectrometry analysis of the DSF molecules produced by this expression system, in addition to those naturally produced by &lt;em&gt;Xf&lt;/em&gt; CoDiRO, are underway.&lt;/p&gt;

&lt;p&gt;Concurrently, a TMV-based vector has been engineered to harbour the same &lt;em&gt;rpfF &lt;/em&gt;gene and induce its transient expression &lt;em&gt;in planta&lt;/em&gt;. Biologically active transcripts of the vector have been inoculated to &lt;em&gt;Nicotiana tabacum&lt;/em&gt; and &lt;em&gt;N. benthamiana&lt;/em&gt; plants, to establish a model system on herbaceous hosts. &lt;em&gt;RpfF&lt;/em&gt; expression was successfully proved by Western blot analysis, whereas movement and systemic colonisation of plant tissues were evaluated by RT-PCR assays. The same viral vector harbouring GFP in replacement of &lt;em&gt;rpfF&lt;/em&gt; is used as a control. Following inoculation with &lt;em&gt;Xf&lt;/em&gt; CoDiRO bacterial cells the system is now being tested to monitor the persistence of DSF expression and its efficacy to lower disease susceptibility or movement of bacterial cells behind the point of inoculation.&lt;/p&gt;</subfield>
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