Structured diversity using EST-PCR and EST-SSR markers in a set of wild blueberry clones and cultivars

Blueberries are perennial, vegetatively propagated out-crossing shrubs with great potential of health benefits. In tetraploid blueberry (Vaccinium spp.), genetic analyses have focused more on diversity, whereas genetic structure has been studied less extensively. This study investigated the genetic structure and diversity in 28 wild clones, six cultivars and two selections of lowbush, half-high and highbush blueberries. Summary statistics, structure estimation and clustering by neighbour-joining (NJ), principal coordinate analysis (PCoA) and by the analysis of molecular variance (AMOVA), using 10 expressed sequence tag-polymerase chain reaction (EST-PCR) and two EST-simple sequence repeat (SSR) primer pairs, were performed to characterize and discriminate the genotypes. A total of 213 markers were detected. Wide genetic diversity was evident from high values of expected heterozygosities, Shannon's index and polymorphism information content, and from AMOVA. Structure analysis subdivided the lowbush blueberries into three distinct groups leaving the half-high and highbush blueberries into one cluster which was in agreement with the NJ clustering and PCoA. Twelve EST-primer pairs efficiently differentiated wild and cultivated blueberries, making this technology valuable for genome mapping and for long-term conservation of blueberry resources for the benefit of germplasm conservation and breeding purposes.