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Int. J. Mol. Sci. 2016, 17(2), 203; doi:10.3390/ijms17020203

Use of Repetitive Sequences for Molecular and Cytogenetic Characterization of Avena Species from Portugal

1
Linking Landscape, Environment, Agriculture and Food (LEAF), Instituto Superior de Agronomia, Universidade de Lisboa, Tapada da Ajuda, 1349-017 Lisboa, Portugal
2
Instituto Nacional de Investigação Agrária e Veterinária, Quinta do Marquês, 2784-505 Oeiras, Portugal
*
Author to whom correspondence should be addressed.
Academic Editor: Jianhua Zhu
Received: 5 November 2015 / Revised: 22 January 2016 / Accepted: 26 January 2016 / Published: 4 February 2016
(This article belongs to the Special Issue Plant Molecular Biology)
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Abstract

Genomic diversity of Portuguese accessions of Avena species—diploid A. strigosa and hexaploids A. sativa and A. sterilis—was evaluated through molecular and cytological analysis of 45S rDNA, and other repetitive sequences previously studied in cereal species—rye subtelomeric sequence (pSc200) and cereal centromeric sequence (CCS1). Additionally, retrotransposons and microsatellites targeting methodologies—IRAP (inter-retrotransposon amplified polymorphism) and REMAP (retrotransposon-microsatellite amplified polymorphism)—were performed. A very high homology was detected for ribosomal internal transcribed sequences (ITS1 and ITS2) between the species analyzed, although nucleolar organizing regions (NOR) fluorescent in situ hybridization (FISH) analysis revealed distinct number of Nor loci between diploid and hexaploid species. Moreover, morphological diversity, evidenced by FISH signals with different sizes, was observed between distinct accessions within each species. pSc200 sequences were for the first time isolated from Avena species but proven to be highly similar in all genotypes analyzed. The use of primers designed for CCS1 unraveled a sequence homologous to the Ty3/gypsy retrotransposon Cereba, that was mapped to centromeric regions of diploid and hexaploid species, being however restricted to the more related A and D haplomes. Retrotransposon-based methodologies disclosed species- and accessions-specific bands essential for the accurate discrimination of all genotypes studied. Centromeric, IRAP and REMAP profiles therefore allowed accurate assessment of inter and intraspecific variability, demonstrating the potential of these molecular markers on future oat breeding programs. View Full-Text
Keywords: Avena Portuguese lines; repetitive sequences; genomic diversity; molecular markers Avena Portuguese lines; repetitive sequences; genomic diversity; molecular markers
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Tomás, D.; Rodrigues, J.; Varela, A.; Veloso, M.M.; Viegas, W.; Silva, M. Use of Repetitive Sequences for Molecular and Cytogenetic Characterization of Avena Species from Portugal. Int. J. Mol. Sci. 2016, 17, 203.

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