Submitted to: Scientia Horticulturae
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/12/2010
Publication Date: 7/26/2010
Citation: Rowland, L.J., Ogden, E.L., Ehlenfeldt, M.K. 2010. EST-PCR markers developed for Highbush Blueberry are also useful for genetic fingerprinting and relationship studies in Rabbiteye Blueberry. Scientia Horticulturae. 125:779-784. Interpretive Summary: Commercial production of blueberry is from multiple species of the genus Vaccinium. About 2/3 of blueberry production is from improved varieties mainly of V. corymbosum (highbush blueberry) and, to a lesser extent, V. virgatum (rabbiteye blueberry). The other 1/3 of blueberry production is from the wild, managed stands of V. angustifolium (lowbush blueberry). Rabbiteye blueberry is not as widely grown as highbush or lowbush blueberry, but its production is important to the southeastern U.S. Until now, there have been no reproducible DNA markers available for use in genetic studies on rabbiteye blueberry. Here we have tested whether genetic markers called EST-PCR markers, originally developed for highbush blueberry, are useful for DNA fingerprinting and relationship studies in rabbiteye blueberry. Our results indicate that the markers were very effective at distinguishing all the rabbiteye varieties in the study and for constructing a family tree. These markers are very easy to generate and affordable; thus, they should have general utility for scientists working on DNA fingerprinting, genetic diversity, and mapping studies in rabbiteye blueberry.
Technical Abstract: Up until now, randomly amplified polymorphic DNA (RAPD) has been the only type of molecular marker used extensively in rabbiteye blueberry (Vaccinium virgatum). Here we have tested whether a type of sequence-tagged site (STS) marker which utilizes specific ~20-mer primers from Expressed Sequence Tags (ESTs) of highbush blueberry (V. corymbosum), which we call EST-PCR markers, are useful for genetic fingerprinting and relationship studies in rabbiteye blueberry. Of 44 EST-PCR primer pairs, from an assortment of genes expressed in flower buds of cold acclimated and non-acclimated plants, and previously shown to amplify polymorphic fragments among a collection of highbush genotypes, 40 (91%) resulted in successful amplification, and 33 of those (83%) amplified polymorphic fragments among the rabbiteye genotypes. The average number of scorable bands per primer pair was two. A dendrogram constructed from genetic similarity values, based on the EST-PCR marker data, tended to group siblings and parent/progeny together, generally agreeing with pedigree information. A group of 20 markers from five EST-PCR primer pairs distinguished all the genotypes in this study. These markers are as easy to generate and as affordable as RAPDs, but are based on actual gene sequences, and should have general utility for DNA fingerprinting, genetic diversity, and mapping studies.