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Development and use of retrotransposons-based markers (IRAP/REMAP) to assess genetic divergence among table grape cultivars

Published online by Cambridge University Press:  18 February 2019

Danuza Kelly Strioto
Affiliation:
Genetics and Breeding, State University of Maringá, Maringá, PR, Brazil
Betty Cristiane Kuhn
Affiliation:
Genetics and Breeding, State University of Maringá, Maringá, PR, Brazil
William Seiji Lemes Nagata
Affiliation:
Biotechnology, State University of Maringá, Maringá, PR, Brazil
Giovana Marinelli
Affiliation:
Biotechnology, State University of Maringá, Maringá, PR, Brazil
Sandra Aparecida Oliveira-Collet
Affiliation:
Departament of Biotechnology, Genetics and Cell Biology, State University of Maringá, Maringá, PR, Brazil
Claudete A. Mangolin
Affiliation:
Departament of Biotechnology, Genetics and Cell Biology, State University of Maringá, Maringá, PR, Brazil
Maria de Fátima P. S. Machado*
Affiliation:
Departament of Biotechnology, Genetics and Cell Biology, State University of Maringá, Maringá, PR, Brazil
*
*Corresponding author. E-mail: mfpsmachado@uem.br

Abstract

For more than four decades after the introduction of cv. Italia (Vitis vinifera L.) in Brazil, several somatic mutations in the genome of cv. Italia and its somatic mutants gave rise to phenotypes which generated at least five new cultivars of fine table grapes. Since no molecular marker proved to be effective in discriminating cv. Italia (V. vinifera L.) and its coloured mutants (Rubi, Benitaka, Brasil, Black Star), primers for the long terminal repeat (LTR) sequences were developed to analyse Inter Retrotransposon Amplified Polymorphism (IRAP) and Retrotransposon-Microsatellite Amplified Polymorphism (REMAP), and investigate how the coloured cultivars derived from clonal propagations of somatic mutations are genetically structured. Primers for LTR sequences of IRAP and REMAP markers were edited from grape sequence databases available at a GenBank. Twenty-four primers, denominated DKS001–DKS024, were edited. Three hundred and forty-nine DNA segments were amplified by individual DKS primers and DKS/ISSR (Inter Simple Sequence Repeats) primer combinations, at an average of 13.96 amplicons per primer pair. High genetic divergence between the five cultivars was inferred from polymorphism in retrotransposons IRAP and REMAP. The analysis of polymorphism of IRAP and REMAP retrotransposons was crucial to show that clonal propagation of somatic mutations may lead towards the formation of genetically divergent cultivars by the formation of genetically structured vineyards and show the mixture of genomes within each cultivar.

Type
Research Article
Copyright
Copyright © NIAB 2019 

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