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Open Access Research article

Development and validation of microsatellite markers for Brachiaria ruziziensis obtained by partial genome assembly of Illumina single-end reads

Pedro IT Silva124, Alexandre M Martins12, Ediene G Gouvea1, Marco Pessoa-Filho3 and Márcio E Ferreira1*

Author Affiliations

1 Embrapa Recursos Genéticos e Biotecnologia, Genetics Lab, PO Box 02372, Brasília, CEP 70770-917, Distrito Federal, Brazil

2 Departamento de Biologia Celular, IB - Universidade de Brasília (UnB) Campus Universitário Darcy Ribeiro, Asa Norte, Brasília, CEP 70910-900, Distrito Federal, Brazil

3 Embrapa Cerrados, PO Box 08223, Brasília, CEP 73310-970, Distrito Federal, Brazil

4 Current address: Dupont Pioneer, Palmas, Tocantins, Brazil

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BMC Genomics 2013, 14:17  doi:10.1186/1471-2164-14-17

Published: 16 January 2013

Abstract

Background

Brachiaria ruziziensis is one of the most important forage species planted in the tropics. The application of genomic tools to aid the selection of superior genotypes can provide support to B. ruziziensis breeding programs. However, there is a complete lack of information about the B. ruziziensis genome. Also, the availability of genomic tools, such as molecular markers, to support B. ruziziensis breeding programs is rather limited. Recently, next-generation sequencing technologies have been applied to generate sequence data for the identification of microsatellite regions and primer design. In this study, we present a first validated set of SSR markers for Brachiaria ruziziensis, selected from a de novo partial genome assembly of single-end Illumina reads.

Results

A total of 85,567 perfect microsatellite loci were detected in contigs with a minimum 10X coverage. We selected a set of 500 microsatellite loci identified in contigs with minimum 100X coverage for primer design and synthesis, and tested a subset of 269 primer pairs, 198 of which were polymorphic on 11 representative B. ruziziensis accessions. Descriptive statistics for these primer pairs are presented, as well as estimates of marker transferability to other relevant brachiaria species. Finally, a set of 11 multiplex panels containing the 30 most informative markers was validated and proposed for B. ruziziensis genetic analysis.

Conclusions

We show that the detection and development of microsatellite markers from genome assembled Illumina single-end DNA sequences is highly efficient. The developed markers are readily suitable for genetic analysis and marker assisted selection of Brachiaria ruziziensis. The use of this approach for microsatellite marker development is promising for species with limited genomic information, whose breeding programs would benefit from the use of genomic tools. To our knowledge, this is the first set of microsatellite markers developed for this important species.