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Direct multiplex PCR for grapevine genotyping and varietal identification

Published online by Cambridge University Press:  05 December 2012

Daniele Migliaro ,
Giacomo Morreale ,
Massimo Gardiman ,
Sara Landolfo  and
Manna Crespan
Daniele Migliaro
Affiliation:
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
Giacomo Morreale
Affiliation:
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
Massimo Gardiman
Affiliation:
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
Sara Landolfo
Affiliation:
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
Manna Crespan*
Affiliation:
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
*
*Corresponding author: E-mail: manna.crespan@entecra.it
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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.

Keywords

Vitismicrosatellitesdirect PCRSSRmultiplex PCRDNA extraction
Type
Short Communication
Information
Plant Genetic Resources , Volume 11 , Issue 2 , August 2013 , pp. 182 - 185
Copyright
Copyright © NIAB 2012 

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