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P-40<br />

Towards a characterization of the PR-10 gene family in grapevine<br />

M. Aless<strong>and</strong>rini, L. Bortesi, A. Lovato, M. Fasoli, G. Zoccatelli, S. Zenoni, V. Trope, M.<br />

Pezzotti, A. Polverari*<br />

Dipartimento di Biotecnologie, Università di Verona, Verona, Italy<br />

*Corresponding author: annalisa.polverari@univr.it<br />

We have started a project aimed at characterizing the expression profile <strong>and</strong> protein activity<br />

of pathogenesis related proteins of the PR-10 family in grapevine. According to a large scale<br />

microarray analysis carried out on a Combimatrix grapevine chip, the expression of several<br />

Tentative consensus annotated as similar to PR-10 proteins is strongly increased in resistant<br />

Vitis riparia, <strong>and</strong> to a lesser extent in V. vinifera, following infection <strong>with</strong> Plasmopara<br />

viticola (Polesani et al., BMC Genomics 2010). PR-10 proteins are induced by pathogen<br />

infection <strong>and</strong> abiotic stresses in a number of species; because they often show<br />

ribonucleolytic activity it has been suggested that they may be involved in antiviral defence;<br />

their antimicrobial activity has been tested <strong>with</strong> controversial results. Up to now, we have<br />

analyzed the expression level of four members of the PR-10 family, predicted in the Vitis<br />

vinifera genome, by Real-time RT-PCR, in response to P. viticola infection at 12 <strong>and</strong> 24<br />

hours post-inoculation (hpi). All are barely detectable in healthy V. vinifera leaves but are<br />

induced, <strong>with</strong> similar levels, in response to infection. An homologous gene, that we called<br />

Vr-PR10, has been isolated from the resistant species Vitis riparia following infection; its<br />

coding sequence has been cloned from V. riparia cDNA <strong>and</strong> the corresponding protein<br />

expressed in E. coli. The purified protein showed ribonucleolytic activity in vitro. Using<br />

antibodies raised against the recombinant protein, which recognize the native protein from<br />

leaf tissue in Western blot analyses, a strong induction of the protein upon infection was<br />

shown. The protein is now being tested for antimicrobial <strong>and</strong> antiviral activity in vitro <strong>and</strong><br />

in vivo.<br />

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