Summary
Large - scale isolation of microsatellite and information in any conifer species is limited. Our knowledge of microsatellite in spruce (Picea spp.) is still sketchy. Genomic libraries of P. mariana were constructed and screened with (AC)15 probes. Over 200 positive clones were found for this dinucleotide and ten were analyzed in details. They were sequenced to confirm the presence of microsatellites and to generate polymerase chain reaction (PCR) primers based on sequences flanking the microsatellites. All sequenced (AC)n clones had repeats of n > 23. Primer pairs were designed from seven sequences. These primer pairs along with 15 primer pairs from white spruce (Picea glauca) were tested on individual trees. Seven primer pairs from P. mariana and three from P. glauca (white spruce) amplified DNA from P. mariana and were used for genetic analysis of black spruce populations from uplands (drylands) and lowlands (wetlands). High levels of polymorphism and heterozygosity were observed in all the populations studied. Both highlands and lowlands showed similar levels of genetic variation. The selected microsatellites sequences are being used for genome organization analysis of black spruce.
References
AMARASINGHE, V. and J. E. CARLSON (2002): The development of microsatellite DNA markers for genetic analysis in Douglas-fir. Can. J. For. Res. 32: 1904-1915.
BOYS, J., M. CHERRY and S. DAYANANDAN (2005): Microsatellite analysis reveals genetically distinct populations of red pine (Pinus resinosa, Pinaceae). American Journal of Botany 92: 833-841.
ECHT, C. S., P. MAY-MARQUARDT, M. HSEIH and R. ZAHORECHAK (1986): Characterization of microsatellites markers in eastern white pine. Genome 39: 1102-1108.
ECHT, C. S., P. MAY-MARQUARDT, M. HSEIH and R. ZAHORECHAK (1996): Characterization of microsatellite markers in eastern white pine. Genome 39: 1102-1108.
ECHT, C. S. (1999): Use of microsatellite markers in management of conifer forest species. In: DOUGLAS, G. C., ed. Strategies for improvement of forest trees. Dublin, Ireland: COFORD National Council for Forest Research and Development: 75-82.
ECHT, C. S., G. G. VENDRAMIN, C. D. NELSON and P. MARQUARDT (1999): Microsatellite DNA as shared genetic markers among conifer species. Can. J. For. Res. 29: 365-371.
EDWARDS, K. J., J. H. A. BARKER, A. DALY, C. JONES and A. KARP (1996): Microsatellite libraries enriched for several microsatellite sequences in plants. BioTechniques 20: 758-760.
GUPTA, A. K., B. Y. KANG, J. K. ROY and O. P. RAJORA (2005): Large scale development of selectively amplified microsatellite polymorphic loci (SAMPL) markers in spruce (Picea). Molecular Ecology notes 5 (3): 481.
HAMILTON, M. B., E. L. PINCUS, A. DI FIORE and R. FLEISHER (1999): Universal linker and ligation procedures for construction of genomic DNA libraries enriched for microsatellites. BioTechniques 27: 500-507.
HODGETTS, R. B., M. A. ALEKSIUK, A. BROWN, C. CLARKE, E. MACDONALD, S. NADEEM and D. KHASA (2001): Development of microsatellite markers for white spruce (Picea glauca) and related species. Theor. Appl. Genet. 102: 1252-1258.
LIU, K. J. and S. V. MUSE (2005): PowerMarker: an integrated analysis environment for genetic marker analysis. Bioinformatics 21: 2128-2129.
MEHES, M., K. K. NKONGOLO and P. MICHAEL (2007): Genetic analysis of Pinus monticola and Pinus strobes using microsatellites. Poster presented at the joint annual meeting of the Canadian Genetic Society of Canada and Canadian Fly Society in Montreal (Qc), June 18-22.
NKONGOLO, K. K., P. MICHAEL and T. DEMERS (2005): Application of ISSR, RAPD, and cytological markers to the certification of Picea mariana, P. glauca, and P. engelmannii trees and their putative hybrids. Genome 48: 302-311.
PFEIFFER, A., A. M. OLVIERI and M. MORGANTE (1997): Identification and characterization of microsatellites in Norway spruce (Picea abies K.). Genome 40: 411-419.
RAJORA, O. P., M. H. RHAMAN, S. DAYANANDAN and A. MOSSELER (2001): Isolation, characterization, inheritance and linkage of microsatellite DNA markers in white spruce and their usefulness in other spruce species. Mol. Gen. Genet. 264: 871-882.
RANGER, M., K. VANDELIGT, P. MICHAEL, K. K. NKONGOLO and P. BECKETT (2007): Microsatellite and ISSR analysis of Pinus banksiana and Pinus resinosa from metal contaminated areas in the Greater Sudbury region. Paper presented at the Mining and Environment IV Conference in Sudbury (ON), October 19-27.
SCOTTI, I., G. P. PAGLIA, F. MAGNI and M. MORGANTE (2002): Efficient development of dinucleotide microsatellite markers in Norway spruce (Picea abies Karst.) through dot-blot selection. Theor. Appl. Genet. 104: 1035-1041.
WANG, Y., J. LUO, X. XUE, H. KORPELAINEN and C. LI (2005): Diversity of microsatellite markers in the populations of Picea asparta originating from the mountains of China. Plant Science 168: 707-714.
YEH, F. C. and T. J. B. BOYLE (1997): Population genetic analysis of co-dominant and dominant markers and quantitative traits. Belg. J. Bot. 129: 157.
