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Combination of high resolution melting (HRM) analysis and SSR molecular markers speeds up plum genotyping: case study genotyping the Greek plum GeneBank collection

Published online by Cambridge University Press:  03 March 2016

Georgios Merkouropoulos*
Affiliation:
Institute of Applied Biosciences, CERTH, Thermi, Thessaloniki 570 01, Greece Hellenic Agricultural Organization ‘Demeter’, Institute of Plant Breeding & Genetic Resources, 57001 Thermi, Greece
Ioannis Ganopoulos
Affiliation:
Institute of Applied Biosciences, CERTH, Thermi, Thessaloniki 570 01, Greece
Athanasios Tsaftaris
Affiliation:
Institute of Applied Biosciences, CERTH, Thermi, Thessaloniki 570 01, Greece Laboratory of Genetics and Plant Breeding, Faculty of Agriculture, Forestry and Natural Environment, School of Agriculture, Aristotle University of Thessaloniki, Thessaloniki 54 124, Greece
Ioannis Papadopoulos
Affiliation:
Technological Educational Institute of Western Macedonia/Branch of Florina, Terma Kontopoulou, 53100 Florina, Greece
Pavlina Drogoudi
Affiliation:
Hellenic Agricultural Organization ‘Demeter’, Institute of Plant Breeding and Genetic Resources, Department of Deciduous Fruit Trees in Naoussa, 38 R.R. Station, 59035, Naoussa, Greece
*
*Corresponding author. E-mail: georgios.merkouropoulos@gmail.com

Abstract

In a germplasm bank collection the conservation and characterization of genetic resources is a prerequisite in order to use the material in breeding projects aiming the creation of new cultivars. In the present study, 54 Prunus salicina domestica and Prunus domestica genotypes (including seven Greek cultivars), maintained in the ex situ National Genebank Collection of Greece, were classified using microsatellite (simple sequence repeat, SSR) markers on high resolution melting (HRM) analysis. The SSR primer pairs were chosen from the published literature as originally designed on Prunus species. This combined approach was used to genotype all plum accessions of the collection highlighting the benefits of either method (HRM and SSRs) for cultivar identification. Dendrograms for P. domestica and P. salicina and a combined one with all the genotypes assayed were produced. A total of 15 from the 19 P. domestica accessions analysed, including all the Greek accessions but ‘Avgati Skopelou’, were grouped into the same clade in the combined dendrogram, whereas the remaining four were dispersed into the P. salinica clades. Bayesian structure analysis confirmed that ‘Avgati Skopelou’ differs from the rest of the Greek plum cultivars since it was not grouped into the same cluster. The combination of HRM and SSRs, provided a considerably faster, cost-effective, closed-tube microsatellite genotyping method for molecular characterization of plum cultivars.

Type
Research Article
Copyright
Copyright © NIAB 2016 

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