Microsatellite marker development and molecular characterization in highbush blueberry (Vaccinium corymbosum L.) and Vaccinium species Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/4f16c609k

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  • Pre-existing Expressed Sequence Tag (EST) libraries (1,305 sequences) and a newly constructed microsatellite-enriched genomic library (136 clones) from the V. corymbosum cultivar 'Bluecrop' were screened for unique Simple Sequence Repeat (SSR)-containing sequences. Our goal was to develop a set of robust SSR markers for use in managing the National Clonal Germplasm Repository blueberry collection. Of 153 microsatellite loci detected in the DNA sequences, 94 primer pairs were tested for amplification and polymorphism in a panel of 12 Vaccinium germplasm accessions. A total of 30 loci (20 EST and 10 genomic) repeatedly amplified a well conserved and highly polymorphic locus of the expected size. The average cross-amplification in 12 Vaccinium species of 44 V. corymbosum-derived SSR that amplified a single locus on agarose gels ranged from 59% in V. macrocarpon Ait. to 95% in diploid V. corymbosum (95% confidence interval for mean 73- 82%). The genetic similarity among 114 accessions of North and South American species of Vaccinium L. were investigated using 5 SSR loci that exhibited high allelic diversity. The genetic similarity measure reproduced many well-accepted taxonomic relationships between species in the genus Vaccinium and show great promise for future use in phylogenetic studies. Within section Cyanococcus, our results supported the recognition of V. elliottii Chapm. as a genetically distinct diploid highbush species and the theory of widespread hybridization among polyploid taxa. A set of 71 accessions representing wild and domesticated highbush blueberry germplasm were genotyped using 28 SSR loci. A total of 627 alleles were detected and produced unique fingerprints for all accessions. The duplication of two accessions being maintained as separate inventories was verified. The highest numbers of taxon-specific alleles were detected in wild V. corymbosum and some cultivars. Genetic similarity measures grouped wild and domesticated blueberries separately. Domesticated blueberries of the southern highbush type formed a group distinct from the northern highbush types. Northern highbush accessions grouped among ancestral clones that have been extensively used in blueberry breeding such as 'Rubel' and 'Stanley'. These SSR markers show excellent promise for future use in germplasm identification, studying wild Vaccinium populations, and linkage mapping.
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