Plant Genetic Resources

Short Communication

Direct multiplex PCR for grapevine genotyping and varietal identification

Daniele Migliaroa1, Giacomo Morrealea1, Massimo Gardimana1, Sara Landolfoa1 and Manna Crespana1 c1

a1 Consiglio per la Ricerca e la Sperimentazione in Agricoltura, Centro di Ricerca per la Viticoltura (CRA-VIT), Viale XXVIII Aprile 26, 31015 Conegliano, Treviso, Italy

Abstract

Grapevine cultivar identification is based mainly on two complementary methodologies: microsatellite (simple sequence repeat (SSR)) DNA analysis and traditional ampelography. Here, we report a direct multiplex PCR approach that allows the simultaneous amplification of 11 SSR loci from crude samples, i.e. bypassing DNA extraction, by using an engineered DNA polymerase improved to tolerate plant PCR inhibitors. Many different plant tissues were successfully amplified: leaf, root, wood, berry flesh and skin, stalk and must, from wine and table grape varieties, and rootstocks. The direct multiplex PCR that we propose is quicker and cheaper than the methodologies used until now, and provides accurate results. Thus, SSR DNA analysis becomes economically more accessible to a larger number of potential users in addition to research institutes.

(Online publication December 05 2012)

Key Words:

  • Vitis ;
  • microsatellites;
  • direct PCR;
  • SSR;
  • multiplex PCR;
  • DNA extraction

Correspondence

c1 Corresponding author: E-mail: manna.crespan@entecra.it

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