PAG-IX: MICROSATELLITE DNA MARKERS IN WHITE SPRUCE (PICEA GLAUCA), AND THEIR COMPATIBILITY AND USEFULNESS IN FIVE OTHER SPRUCE SPECIES

PAG-IX   Plant & Animal Genome IX Conference

Town & Country Hotel, San Diego, CA, January 13-17, 2001.

Workshop: Forest Tree Genome Mapping
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MICROSATELLITE DNA MARKERS IN WHITE SPRUCE (PICEA GLAUCA), AND THEIR COMPATIBILITY AND USEFULNESS IN FIVE OTHER SPRUCE SPECIES

OM P. RAJORA1, Muhammad H. Rahman2, Alex Mosseler3,

1 Department of Biology, Life Sciences Centre, Dalhousie University, Halifax, NS B3H 4J1, Canada
2 Department of Renewable Resources, University of Alberta, Edmonton, AB T6G 2H1, Canada
3 Canadian Forest Service, Atlantic Forestry Centre, PO Box 4000, Fredericton, NB E3B 5P7, Canada

Microsatellite DNA/simple sequence repeat (SSR) loci were identified, isolated and characterized in white spruce (Picea glauca) by screening both non-enriched and (AG/TC)n -enriched partial genomic libraries. The compatibility and usefulness of the P. glauca microsatellite DNA markers were determined in five other Picea species. Twenty-four microsatellites were identified by sequencing 32 clones selected from screening of 5400 clones from two libraries. The (AG/TC)n microsatellites were the most abundant in the non-enriched library. Eight microsatellite DNA loci were of single-copy type, six of which were polymorphic. Eighty-seven alleles were detected at the six polymorphic SSR loci in 32 P. glauca individuals. The number of alleles at the polymorphic SSR loci ranged from 2 to 22, with an average of 14.5 alleles per locus, and the observed heterozygosity ranged from 0.48 to 0.91, with a mean of 0.66 per locus. Microsatellite DNA variants at each of the five SSR loci informative for inheritance studies followed a single-locus codominant Mendelian inheritance. Joint two-locus segregation tests indicated a complete linkage between two loci and no linkage between any of the remaining SSR loci. Each of the 32 P. glauca individuals examined had unique single or two-locus genotypes. With one exception, primer pairs for all six polymorphic SSR loci were successful in PCR-amplifying genomic DNA and resolving polymorphic microsatellites of comparable size in P. engelmanni, P. sitchensis, P. mariana, P. rubens, and P. abies. The closely related P. mariana and P. rubens, and P. glauca and P. sitchensiss could be distinguished by the markers of one SSR locus. The microsatellite DNA markers developed and reported here could be used for assisting various genetics, breeding, biotechnology, tree forensics, genome mapping, conservation, restoration, and sustainable forest management programs in spruce species.

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