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P-41<br />

Assessing the genetic variability of grape clones<br />

S. Vezzulli* 1 , L. Leonardelli 1 , U. Malossini 2 , M. Stefanini 1 , R. Velasco 1 , C. Moser 1<br />

1<br />

IASMA Research <strong>and</strong> Innovation Centre, Fondazione Edmund Mach, Genomics <strong>and</strong> Crop<br />

Biology Area, San Michele a/Adige, Trento, Italy; 2 IASMA Centre for Technology Transfer,<br />

Fondazione Edmund Mach, San Michele a/Adige, Trento, Italy<br />

*Corresponding author: silvia.vezzulli@iasma.it<br />

Grapevine (Vitis vinifera L.) is a long-living <strong>and</strong> woody plant grown worldwide. Its perennial<br />

nature <strong>and</strong> vegetative propagation over long periods favours accumulation <strong>and</strong> fixing of<br />

mutations <strong>with</strong>in individual genotypes, which might exhibit altered phenotypes. Clonal selection,<br />

as a procedure of crop improvement, takes advantage of the identification of sports <strong>with</strong><br />

agronomically <strong>and</strong> enologically important traits, which are vegetatively propagated to raise new<br />

grape clones. Given the high economical value, as reflected by related patenting <strong>and</strong> legal issues,<br />

their identification is of great relevance. While cultivar identification in grapevines is<br />

traditionally based on ampelographic descriptors <strong>and</strong> on reference microsatellite (SSR) profiles,<br />

clone discrimination is not possible <strong>with</strong> such tools. To allow true genetic identification of<br />

clones, new specific molecular markers have to be implemented. Given the availability of the<br />

Pinot Noir (clone ENTAV 115) genome sequence, it is timely to use techniques based on the<br />

polymorphism information of specific genomic sequences of interest, such as DNA transposons<br />

<strong>and</strong> retrotransposons. Transposable elements (TEs), which possess the capability to change their<br />

genomic location, are indeed potential source of mutations leading to clonal variation. An<br />

alternative way to assess clonal genetic diversity consists of the study of various polymorphisms<br />

at different layers; clonal mutations can occur either at both L1 <strong>and</strong> L2 layers or at a single layer.<br />

In this work, to fathom clonal genetic variability <strong>with</strong>in six wine grape cultivars, we adopted<br />

three different methods. Firstly, we have carried out a Transposon Display analysis on 141 grape<br />

genotypes, which correspond to 47 clones (both registered <strong>and</strong> biotypes) belonging to the Pinot<br />

Noir, Pinot Gris, Pinot Blanc, Meunier, Teroldego <strong>and</strong> Gewürztraminer cultivars in 3 biological<br />

replicates (plants). We have tested them using 17 primers targeting specific regions (LTRs, LTR<br />

downstream or upstream, ORF) of 6 different TE families. Secondly, by using 6 triallelic SSR<br />

markers we have analysed four tissues (leaf, berry skin, flesh <strong>and</strong> root) of the 19 registered<br />

clones. Finally, we have genotyped the same set of samples by SNPlex TM Genotyping System,<br />

testing about 500 electronic SNPs identified in coding <strong>and</strong> non-coding regions of the mentioned<br />

grape genome. Based on SNPlex results, we are currently resequencing the regions <strong>with</strong> a high<br />

level of polymorphism to confirm the presence of clonal mutations. Here we report the results of<br />

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