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Title: UTILITY OF BLUEBERRY-DERIVED EST-PCR PRIMERS IN RELATED ERICACEAE SPECIES

Author
item Rowland, Lisa
item DHANARAJ, ANIK - VISITING SCIENTIST
item Polashock, James
item ARORA, RAJEEV - IOWA STATE UNIVERSITY

Submitted to: HortScience
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/9/2003
Publication Date: 12/20/2003
Citation: Rowland, L.J., Dhanaraj, A., Polashock, J.J., Arora, R. 2003. Utility of blueberry-derived est-pcr primers in related ericaceae species. Hortscience. 38:1428-1432.

Interpretive Summary: DNA markers are useful in genetic studies of plants for distinguishing (fingerprinting) cultivars, determining genetic relatedness, and for mapping purposes. Recently, we developed a type of DNA marker for blueberry based on sequences of expressed genes called EST-PCR (expressed sequence tag-polymerase chain reaction) markers. These markers were tested with a variety of different blueberry plants and they proved very effective at DNA fingerprinting and mapping. Although there are many advantages to using EST-PCR markers, they are quite expensive, in terms of time and resources, to produce. If the EST-PCR markers designed for one species can be used in other related species, then the cost involved in developing these markers for the other species is significantly reduced. Here, we have tested whether these blueberry-derived markers are useful in other species of the Ericaceae family, closely related cranberry species and more distantly related rhododendron species. Approximately 90% of the blueberry-derived markers proved useful in cranberry and approximately 75% were useful in rhododendron. Thus, these markers will be available to scientists for use in breeding new cranberry and rhododendron cultivars and to scientists, breeders, and the nursery industry for DNA fingerprinting of cultivars.

Technical Abstract: Expressed sequence tag-polymerase chain reaction (EST-PCR) markers for DNA fingerprinting and mapping in blueberry (Vaccinium spp) had previously been developed from expressed sequence tags (ESTs) produced from a cDNA library, derived from RNA from floral buds of cold acclimated plants. Because EST-PCR markers are derived from gene coding regions, they are more likely to be conserved across populations and species than markers derived from random regions of DNA, such as randomly amplified polymorphic DNA (RAPD) or amplified fragment length polymorphism (AFLP) markers. In this study, we tested whether many of the EST-PCR primer pairs developed for blueberry are capable of amplifying DNA fragments in other members of the family Ericaceae. Closely related cranberry genotypes (two wild selections of V. oxycoccus L. and two cultivars of V. macrocarpon Aiton, `Early Black and `Stevens) and more distantly related rhododendron genotypes (one wild selection each of Rhododendron arboreum Marsh, R. maximum L., and R. ponticum L. and three complex species hybrids, `Sonata, `Grumpy Yellow, and `Roseum elegans) were used. Of 26 primer pairs tested in cranberry, 23 (89%) resulted in successful amplification and eight of those (35%) amplified polymorphic fragments. Of 39 primer pairs tested in rhododendron, 29 (74%) resulted in successful amplification and 21 of those (72%) amplified polymorphic fragments. Thus, these markers should be useful for DNA fingerprinting, comparative mapping, and assessing genetic diversity within species of the genus Vaccinium and between some genera of the family Ericaceae.