Skip to main content

Direct multiplex PCR for grapevine genotyping and varietal identification

  • Daniele Migliaro (a1), Giacomo Morreale (a1), Massimo Gardiman (a1), Sara Landolfo (a1) and Manna Crespan (a1)...

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.

Corresponding author
*Corresponding author: E-mail:
Hide All
Bowers JE, Dangl GS and Meredith CP (1999) Development and characterization of additional microsatellite DNA markers for grape. American Journal of Enology and Viticulture 50: 243246.
Crespan M (2003) The parentage of Muscat of Hamburg. Vitis 42: 193197.
Crespan M, Cabello F, Giannetto S, Ibáñez J, Kontić JK, Maletić E, Pejić I, Rodriguez I and Antonacci D (2006) Malvasia delle Lipari, Malvasia di Sardegna, Greco di Gerace, Malvasia de Sitges and Malvasia dubrovačka – synonyms of an old and famous grape cultivar. Vitis 45: 6973.
Ibáñez J, Vargas AM, Palancar M, Borrego J and De Andrés MT (2009) Genetic relationships among table-grape varieties. American Journal of Enology and Viticulture 60: 3542.
Kalinowski ST, Taper ML and Marshall TC (2007) Revising how the computer program CERVUS accommodates genotyping error increases success in paternity assignment. Molecular Ecology 16: 10991106.
Merdinoglu D, Butterlin G, Bevilacqua L, Chiquet V, Adam-Blondon AF and Decroocq S (2005) Development and characterization of a large set of microsatellite markers in grapevine (Vitis vinifera L.) suitable for multiplex PCR. Molecular Breeding 15: 349366.
Moreno-Sanz P, Loureiro MD and Suárez B (2011) Microsatellite characterization of grapevine (Vitis vinifera L.) genetic diversity in Asturias (Northern Spain). Scientia Horticulturae 129: 433440.
Organisation Internationale de la Vigne et du Vin (OIV) (2009) OIV Descriptor List for Grape Varieties and Vitis Species, 2nd edn. Available at: Paris: OIV Publisher.
Sefc KM, Lefort F, Grando MS, Scott KD, Steinkellner H and Thomas MR (2001) Microsatellite markers for grapevine: a state of the art. In: Roubelakis-Angelakis (ed.) Molecular Biology and Biotechnology of Grapevine. Amsterdam: Kluwer Publishers, pp. 130.
This P, Jung A, Boccacci P, Borrego J, Botta R, Costantini L, Crespan M, Dangl GS, Eisenheld C, Ferreira-Monteiro F, Grando S, Ibáñez J, Lacombe T, Laucou V, Magalhães M, Meredith CP, Milani N, Peterlunger E, Regner F, Zulini L and Maul E (2004) Development of a standard set of microsatellite reference alleles for identification of grape cultivars. Theoretical and Applied Genetics 109: 14481458.
This P, Lacombe T and Thomas MR (2006) Historical origins and genetic diversity of wine grapes. Trends in Genetics 22: 511519.
Welter LJ, Göktürk-Baydar N, Akkurt M, Maul E, Eibach R, Töpfer R and Zyprian EM (2007) Genetic mapping and localization of quantitative trait loci affecting fungal disease resistance and leaf morphology in grapevine (Vitis vinifera L.). Molecular Breeding 20: 359374.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Plant Genetic Resources
  • ISSN: 1479-2621
  • EISSN: 1479-263X
  • URL: /core/journals/plant-genetic-resources
Please enter your name
Please enter a valid email address
Who would you like to send this to? *


Type Description Title
Supplementary Materials

Migliaro Supplementary Material
Tables 1-2

 Word (53 KB)
53 KB


Full text views

Total number of HTML views: 9
Total number of PDF views: 33 *
Loading metrics...

Abstract views

Total abstract views: 285 *
Loading metrics...

* Views captured on Cambridge Core between September 2016 - 23rd November 2017. This data will be updated every 24 hours.