Graduate Thesis Or Dissertation

 

Characterization of European hazelnut (Corylus avellana) cultivars using SSR markers Pubblico Deposited

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  • Twenty-one pairs of simple sequence repeat (SSR) primers were used to investigate genetic diversity in 270 accessions of European hazelnut (Corylus avellana) representing a wide geographic range. A capillary electrophoresis system and ABI Genescan® and Genotyper® software were used to determine the allele size generated from each PCR reaction based on an internal lane standard. A total of 211 alleles were amplified and the number of alleles detected ranged from 5 to 15 per locus with an average of 10.05. The observed heterozygosity (H₀) for individual loci ranged from 0.24 to 0.88, with an average of 0.67 over all loci. The presence of null alleles was detected at three loci (CAC-C010, CaT-C504 and CaT-B508) by comparing the pedigree information of some cultivars. PowerMarker software was used to generate a genetic similarity matrix based on possible pair-wise combinations of accessions using the "proportion of shared alleles". UPGMA cluster analysis was used to construct a phenogram from the genetic similarity matrix using PowerMarker and MEGA3 software. The phenogram revealed geographically tight clusters and some synonyms among European hazelnut cultivars. Of the 274 accessions in the population, 200 are unique cultivars, 70 are suspected synonym accessions (different trees), as they are morphologically identical and 4 are duplicate DNA templates from the same tree included as checks. Of the 211 total alleles amplified, 22 were unique, as they were detected only in one cultivar. Nine of the unique alleles were amplified in cultivars that fell outside of the tightly clustered cultivar groups. A subset of 11 loci is recommended for in future hazelnut fingerprinting studies. A total of 144 seedlings from a controlled cross of OSU 252.146 x OSU 414.062 were scored for 33 SSR markers and 29 of them were successfully integrated into the RAPD marker-based hazelnut linkage map constructed using the two-way pseudotestcross approach, where they will serve as "anchor loci". Two loci showed aberrant segregation ratios and two loci remained unlinked. Eleven linkage groups were identified for each parent, corresponding to the haploid chromosomes number of hazelnut (2n=2x=22) and spaning a total distance of 668 cM in the susceptible parent and 813 cM in the resistant parent. The order of homologous SSR loci in the two parents was collinear in most cases. Placement of these SSR "anchor loci" on the hazelnut linkage map will make it useful in other populations.
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