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ARS Home » Pacific West Area » Corvallis, Oregon » National Clonal Germplasm Repository » Research » Publications at this Location » Publication #333837

Research Project: Management of Temperate-Adapted Fruit, Nut, and Specialty Crop Genetic Resources and Associated Information

Location: National Clonal Germplasm Repository

Title: A multiplexed microsatellite fingerprinting set for hazelnut cultivar identification

Author
item Akin, Meleksen - Oregon State University
item Nyberg, April
item Postman, Joseph
item Mehlenbacher, Shawn - Oregon State University
item Bassil, Nahla

Submitted to: European Journal of Horticultural Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/21/2016
Publication Date: 12/1/2016
Citation: Akin, M., Nyberg, A.M., Postman, J.D., Mehlenbacher, S., Bassil, N.V. 2016. A multiplexed microsatellite fingerprinting set for hazelnut cultivar identification. European Journal of Horticultural Science. 81(6):327-338.

Interpretive Summary: The objective of this study was to develop a cost-effective DNA test for hazelnuts. We tested 20 DNA regions and identified 14 that resulted in fragments that are easy to distinguish with a computer program. This DNA test was used to genotype 102 hazelnut trees from different origins. It distinguished unique genotypes mainly according to parentage and in some cases based on geographic origin. It identified each of the cultivars released from the Oregon State University Breeding Program and confirmed parentage in six cultivars. Tools for DNA fingerprinting of clonally propagated horticultural crops like hazelnut are in demand and this test constitutes a reliable, less-time consuming and cost-effective procedure for identity and parentage confirmation in hazelnut.

Technical Abstract: The objective of this study was to develop a robust and cost-effective fingerprinting set for hazelnuts using microsatellite (SSR) markers. Twenty SSRs containing repeat motifs of = three nucleotides distributed throughout the hazelnut genome were screened on eight genetically diverse cultivars to assess polymorphism, allele size range, and ease of scoring. Six SSRs were discarded after genotyping 96 hazelnut samples either due to large allele bin widths and/or alleles that do not match the motifs complicating allele scoring. Fourteen polymorphic, easy-to-score SSRs were selected and amplified in a single multiplex. The 14-SSR multiplexed set generated the same alleles that were obtained when amplifying each SSR individually in the eight test accessions. SSR primer concentrations were then optimized to generate a clear signal for each locus. This 14-SSR fingerprinting set was used to genotype 102 hazelnut accessions from different origins. The fingerprinting set distinguished unique accessions mainly according to parentage and in some cases based on geographic origin. They identified each of the cultivars released from the Oregon state University breeding program and confirmed parentage in six cultivars. Tools for DNA fingerprinting of clonally propagated horticultural crops like hazelnut are in demand and this multiplexed set constitutes a reliable, less-time consuming and cost-effective procedure for identity and parentage confirmation in hazelnut.