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Development, characterization and inheritance of new microsatellites in olive (Olea europaea L.) and evaluation of their usefulness in cultivar identification and genetic relationship studies

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Abstract

Twelve new microsatellites have been developed in olive. For that purpose, a genomic library of the olive cultivar ‘Arbequina’ was enriched for GA, GT and ACT repeats. Two methods of screening yielded 27 sequences containing microsatellites out of the 119 clones sequenced. The GA repeat seems to be the most abundant motif. Among sequences containing microsatellites, 4 (14.8%) were redundant, 1 (3.7%) was previously described in the literature and 12 (44.4%) could not be used for primers design because the repeat motifs were incomplete. Suitable primer pairs were obtained for the remaining 10 (37.0%) sequences plus an additional 14 recovered from a formerly developed library. For the 24 primer pairs designed, 4 failed to amplify, 8 produced a complex bands pattern and 12 succeeded in giving amplification products. Considering these 12 primer pairs, 10 showed single locus amplification, whereas the other 2 revealed two loci each. This was demonstrated by studying allele segregation in two olive progenies. Sixty-eight alleles were detected for the 12 microsatellites when 51 olive cultivars were analysed. The number of alleles per locus ranged from 1 to 13. The expected heterozygosity varied between 0 and 0.83. All pairs of cultivars could be distinguished using only three microsatellites due to their great discrimination power value. The data coming from genotyping the 51 olive cultivars for 7 out of the 12 new microsatellites were used for constructing a dendrogram by unweighted pair group method with arithmetic mean cluster analysis using the Dice similarity coefficient. Cultivar association according to their geographical origin was observed.

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Acknowledgements

The assistance of Dr. Dorado in the design of primers is acknowledged with gratitude. The authors are grateful to the World Olive Germplasm Bank of CIFA ‘Alameda del Obispo’ in Córdoba (Spain) for the easy access to the plant material. This research was supported by the ‘CAO00-018-C7-1’ project of the Spanish Ministry of Agriculture, Food and Fisheries and by the European OLIV-TRACK project. Aurora Díaz is grateful for the granting of a predoctoral fellowship to ‘Junta de Andalucía’.

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Authors and Affiliations

  1. Instituto de Agricultura Sostenible-CSIC, Alameda del Obispo s/n, 14080, Córdoba, Spain

    Aurora Díaz & Antonio Martín

  2. IFAPA, CICE, Junta de Andalucía, Avda Menéndez Pidal s/n, 14080, Córdoba, Spain

    Raúl De la Rosa

  3. Departamento de Ciencias Agroforestales, University of Sevilla-EUITA, Ctra Utrera km 1, 41013, Sevilla, Spain

    Pilar Rallo

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  1. Aurora Díaz
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  2. Raúl De la Rosa
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  3. Antonio Martín
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  4. Pilar Rallo
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Correspondence to Aurora Díaz.

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Díaz, A., De la Rosa, R., Martín, A. et al. Development, characterization and inheritance of new microsatellites in olive (Olea europaea L.) and evaluation of their usefulness in cultivar identification and genetic relationship studies. Tree Genetics & Genomes 2, 165–175 (2006). https://doi.org/10.1007/s11295-006-0041-5

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