Eur.J.Hortic.Sci. 81 (6) 327-338 | DOI: 10.17660/eJHS.2016/81.6.6|
ISSN 1611-4426 print and 1611-4434 online | © ISHS 2016 | European Journal of Horticultural Science | Original article
A multiplexed microsatellite fingerprinting set for hazelnut cultivar identification
M. Akin1, A. Nyberg2, J. Postman2, S. Mehlenbacher1 and N.V. Bassil2
1Oregon State University, Department of Horticulture, Corvallis, OR 97331, USA
2USDA-ARS, National Clonal Germplasm Repository, Corvallis, OR 97333, USA
The objective of this study was to develop a robust and cost-effective fingerprinting set for hazelnuts using microsatellite markers, also known as simple sequence repeat (SSR) markers. Twenty SSRs containing repeat motifs of three or more nucleotides distributed throughout the hazelnut genome were screened on eight 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, which complicates 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 the parentage of 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.
Corylus avellana L., genetic resources, identity, markers, simple sequence repeats, trueness-to-type
Significance of this study
What is already known on this subject?
What are the new findings?
Microsatellite markers are available in hazelnut and can distinguish hazelnut cultivars.
What is the expected impact on horticulture?
We found that microsatellite markers with a minimum of three core repeats have fewer artifacts than the more abundant dinucleotide-containing markers. After evaluating 20 SSRs in hazelnut, we developed a DNA test that contains 14 such SSRs and that can be amplified in a single PCR reaction. This DNA test distinguished all unique hazelnut cultivars tested with the exception of clonal duplicates in the field or in tissue culture.
This DNA test provides hazelnut growers, nurserymen, propagators, breeders, curators of collections and other scientists with an economical and reliable tool to confirm identity and parentage, detect propagation errors and enforce intellectual property.
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Received: 20 September 2016 | Accepted: 21 November 2016 | Published: 23 December 2016 | Available online: 23 December 2016